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Sample records for regulates filament formation

  1. Metabolic regulation via enzyme filamentation

    PubMed Central

    Aughey, Gabriel N.; Liu, Ji-Long

    2016-01-01

    Abstract Determining the mechanisms of enzymatic regulation is central to the study of cellular metabolism. Regulation of enzyme activity via polymerization-mediated strategies has been shown to be widespread, and plays a vital role in mediating cellular homeostasis. In this review, we begin with an overview of the filamentation of CTP synthase, which forms filamentous structures termed cytoophidia. We then highlight other important examples of the phenomenon. Moreover, we discuss recent data relating to the regulation of enzyme activity by compartmentalization into cytoophidia. Finally, we hypothesize potential roles for enzyme filament formation in the regulation of metabolism, development and disease. PMID:27098510

  2. Molecular Mechanisms Regulating Cell Fusion and Heterokaryon Formation in Filamentous Fungi.

    PubMed

    Daskalov, Asen; Heller, Jens; Herzog, Stephanie; Fleißner, André; Glass, N Louise

    2017-03-01

    For the majority of fungal species, the somatic body of an individual is a network of interconnected cells sharing a common cytoplasm and organelles. This syncytial organization contributes to an efficient distribution of resources, energy, and biochemical signals. Cell fusion is a fundamental process for fungal development, colony establishment, and habitat exploitation and can occur between hyphal cells of an individual colony or between colonies of genetically distinct individuals. One outcome of cell fusion is the establishment of a stable heterokaryon, culminating in benefits for each individual via shared resources or being of critical importance for the sexual or parasexual cycle of many fungal species. However, a second outcome of cell fusion between genetically distinct strains is formation of unstable heterokaryons and the induction of a programmed cell death reaction in the heterokaryotic cells. This reaction of nonself rejection, which is termed heterokaryon (or vegetative) incompatibility, is widespread in the fungal kingdom and acts as a defense mechanism against genome exploitation and mycoparasitism. Here, we review the currently identified molecular players involved in the process of somatic cell fusion and its regulation in filamentous fungi. Thereafter, we summarize the knowledge of the molecular determinants and mechanism of heterokaryon incompatibility and place this phenomenon in the broader context of biotropic interactions and immunity.

  3. Reversible S-glutathionylation of Cys 374 regulates actin filament formation by inducing structural changes in the actin molecule.

    PubMed

    Dalle-Donne, I; Giustarini, D; Rossi, R; Colombo, R; Milzani, A

    2003-01-01

    S-glutathionylation, the reversible formation of mixed disulphides of cysteinyl residues in target proteins with glutathione, occurs under conditions of oxidative stress; this could be a posttranslational mechanism through which protein function is regulated by the cellular redox status. A novel physiological relevance of actin polymerization regulated by glutathionylation of Cys(374) has been recently suggested. In the present study we showed that glutathionylated actin (GS-actin) has a decreased capacity to polymerize compared to native actin, filament elongation being the polymerization step actually inhibited. Actin polymerizability recovers completely after dethiolation, indicating that S-glutathionylation does not induce any protein denaturation and is therefore a reversible oxidative modification. The increased exposure of hydrophobic regions of protein surface observed upon S-glutathionylation indicates changes in actin conformation. Structural alterations are confirmed by the increased rate of ATP exchange as well as by the decreased susceptibility to proteolysis of the subtilisin cleavage site between Met(47) and Gly(48), in the DNase-I-binding loop of the actin subdomain 2. Structural changes in the surface loop 39-51 induced by S-glutathionylation could influence actin polymerization in view of the involvement of the N-terminal portion of this loop in intermonomer interactions, as predicted by the atomic models of F-actin.

  4. Simulations of Filament Channel Formation

    NASA Astrophysics Data System (ADS)

    Knizhnik, K. J.; Antiochos, S. K.; DeVore, C. R.

    2014-12-01

    A major unexplained feature of the solar atmosphere is the accumulation of magnetic shear, in the form of filament channels, at photospheric polarity inversion lines (PILs). In addition to free energy, this shear also represents magnetic helicity, which is conserved under reconnection. In this work, we address the problem of filament channel formation and show how they acquire their shear and magnetic helicity. Results of 3D MHD simulations using the Adaptively Refined MHD Solver (ARMS) are presented that support the magnetic helicity-condensation model of filament-channel formation described by Antiochos, 2013. We consider the supergranular twisting of a quasi-potential flux system that is bounded by a PIL and contains a coronal hole (CH). The magnetic helicity injected by the small-scale photospheric motions is shown to inverse-cascade up to the largest allowable scales that define the closed flux system: the PIL and the CH boundary. This process produces field lines that are both sheared and smooth, and are sheared in opposite senses at the PIL and the CH. The accumulated helicity and shear flux are shown to be in excellent quantitative agreement with the helicity-condensation model. We present a detailed analysis of the simulation, including comparisons of our analytical and numerical results, and discuss their implications for observations.

  5. Dynamics of heteromolecular filament formation

    NASA Astrophysics Data System (ADS)

    Dear, Alexander J.; Michaels, Thomas C. T.; Knowles, Tuomas P. J.

    2016-11-01

    The self-assembly of molecular building blocks into linear filaments is a common form of self-organization in nature and underlies the formation of supra-molecular polymers in a variety of contexts, including in both functional and aberrant biology. To date, attention has focused mainly on homomolecular assembly phenomena; however, it has recently become apparent that heteromolecular assemblies can be common, and, for instance, pathological protein filaments such as amyloid aggregates form in vivo in environments supporting copolymerization. Here, we present a general kinetic scheme for heteromolecular filament formation and derive closed-form analytical expressions that describe the dynamics of such systems. Our results reveal the existence of a demixing transition time controlled by the relative rates of depletion of the different aggregating species, after which predominantly homomolecular polymers are formed even when the initial solution is heteromolecular. Furthermore, these results may be applied to the analysis of experimental kinetic data on the aggregation of mixtures of proteins, to determine which fundamental reaction steps occur between unlike proteins, and to provide accurate estimates of their rate constants.

  6. Filament-Filament Switching Can Be Regulated by Separation Between Filaments Together with Cargo Motor Number

    PubMed Central

    Erickson, Robert P.

    2013-01-01

    How intracellular transport controls the probability that cargos switch at intersections between filaments is not well understood. In one hypothesis some motors on the cargo attach to one filament while others attach to the intersecting filament, and the ensuing tug-of-war determines which filament is chosen. We investigate this hypothesis using 3D computer simulations, and discover that switching at intersections increases with the number of motors on the cargo, but is not strongly dependent on motor number when the filaments touch. Thus, simply controlling the number of active motors on the cargo cannot account for in vivo observations that found reduced switching with increasing motor number, suggesting additional mechanisms of regulation. We use simulations to show that one possible way to regulate switching is by simultaneously adjusting the separation between planes containing the crossing filaments and the total number of active motors on the cargo. Heretofore, the effect of filament-filament separation on switching has been unexplored. We find that the switching probability decreases with increasing filament separation. This effect is particularly strong for cargos with only a modest number of motors. As the filament separation increases past the maximum head-to-head distance of the motor, individual motors walking along a filament will be unable to reach the intersecting filament. Thus, any switching requires that other motors on the cargo attach to the intersecting filament and haul the cargo along it, while motor(s) engaged on the original filament detach. Further, if the filament separation is large enough, the cargo can have difficulty proceeding along the initial filament because the engaged motors can walk underneath the intersecting filament, but the cargo itself cannot fit between the filaments. Thus, the cargo either detaches entirely from the original filament, or must dip to the side of the initial filament and then pass below the crossing

  7. The Regulation of Filamentous Growth in Yeast

    PubMed Central

    Cullen, Paul J.; Sprague, George F.

    2012-01-01

    Filamentous growth is a nutrient-regulated growth response that occurs in many fungal species. In pathogens, filamentous growth is critical for host–cell attachment, invasion into tissues, and virulence. The budding yeast Saccharomyces cerevisiae undergoes filamentous growth, which provides a genetically tractable system to study the molecular basis of the response. Filamentous growth is regulated by evolutionarily conserved signaling pathways. One of these pathways is a mitogen activated protein kinase (MAPK) pathway. A remarkable feature of the filamentous growth MAPK pathway is that it is composed of factors that also function in other pathways. An intriguing challenge therefore has been to understand how pathways that share components establish and maintain their identity. Other canonical signaling pathways—rat sarcoma/protein kinase A (RAS/PKA), sucrose nonfermentable (SNF), and target of rapamycin (TOR)—also regulate filamentous growth, which raises the question of how signals from multiple pathways become integrated into a coordinated response. Together, these pathways regulate cell differentiation to the filamentous type, which is characterized by changes in cell adhesion, cell polarity, and cell shape. How these changes are accomplished is also discussed. High-throughput genomics approaches have recently uncovered new connections to filamentous growth regulation. These connections suggest that filamentous growth is a more complex and globally regulated behavior than is currently appreciated, which may help to pave the way for future investigations into this eukaryotic cell differentiation behavior. PMID:22219507

  8. Filament Channel Formation via Magnetic Helicity Condensation

    NASA Astrophysics Data System (ADS)

    Knizhnik, K. J.; Antiochos, S. K.; DeVore, C. R.

    2015-08-01

    A major unexplained feature of the solar atmosphere is the accumulation of magnetic shear in the form of filament channels at photospheric polarity inversion lines (PILs). In addition to free energy, this shear represents magnetic helicity, which is conserved under reconnection. In this paper we address the problem of filament channel formation and show how filaments acquire their shear and magnetic helicity. The results of three-dimensional (3D) simulations using the Adaptively Refined Magnetohydrodynamics Solver are presented. Our findings support the model of filament channel formation by magnetic helicity condensation that was developed by Antiochos. We consider the small-scale photospheric twisting of a quasi-potential flux system that is bounded by a PIL and contains a coronal hole (CH). The magnetic helicity injected by the small-scale photospheric motions is shown to inverse cascade up to the largest allowable scales that define the closed flux system: the PIL and the CH. This process produces field lines that are both sheared and smooth, and are sheared in opposite senses at the PIL and the CH. The accumulated helicity and shear flux are shown to be in excellent quantitative agreement with the helicity condensation model. We present a detailed analysis of the simulations, including comparisons of our analytical and numerical results, and discuss their implications for observations.

  9. FILAMENT CHANNEL FORMATION VIA MAGNETIC HELICITY CONDENSATION

    SciTech Connect

    Knizhnik, K. J.; Antiochos, S. K.; DeVore, C. R.

    2015-08-20

    A major unexplained feature of the solar atmosphere is the accumulation of magnetic shear in the form of filament channels at photospheric polarity inversion lines (PILs). In addition to free energy, this shear represents magnetic helicity, which is conserved under reconnection. In this paper we address the problem of filament channel formation and show how filaments acquire their shear and magnetic helicity. The results of three-dimensional (3D) simulations using the Adaptively Refined Magnetohydrodynamics Solver are presented. Our findings support the model of filament channel formation by magnetic helicity condensation that was developed by Antiochos. We consider the small-scale photospheric twisting of a quasi-potential flux system that is bounded by a PIL and contains a coronal hole (CH). The magnetic helicity injected by the small-scale photospheric motions is shown to inverse cascade up to the largest allowable scales that define the closed flux system: the PIL and the CH. This process produces field lines that are both sheared and smooth, and are sheared in opposite senses at the PIL and the CH. The accumulated helicity and shear flux are shown to be in excellent quantitative agreement with the helicity condensation model. We present a detailed analysis of the simulations, including comparisons of our analytical and numerical results, and discuss their implications for observations.

  10. Filament Channel Formation, Eruption, and Jet Generation

    NASA Astrophysics Data System (ADS)

    DeVore, C. Richard; Antiochos, Spiro K.; Karpen, Judith T.

    2017-08-01

    The mechanism behind filament-channel formation is a longstanding mystery, while that underlying the initiation of coronal mass ejections and jets has been studied intensively but is not yet firmly established. In previous work, we and collaborators have investigated separately the consequences of magnetic-helicity condensation (Antiochos 2013) for forming filament channels (Zhao et al. 2015; Knizhnik et al. 2015, 2017a,b) and of the embedded-bipole model (Antiochos 1996) for generating reconnection-driven jets (Pariat et al. 2009, 2010, 2015, 2016; Wyper et al. 2016, 2017). Now we have taken a first step toward synthesizing these two lines of investigation. Our recent study (Karpen et al. 2017) of coronal-hole jets with gravity and wind employed an ad hoc, large-scale shear flow at the surface to introduce magnetic free energy and form the filament channel. In this effort, we replace the shear flow with an ensemble of local rotation cells, to emulate the Sun’s ever-changing granules and supergranules. As in our previous studies, we find that reconnection between twisted flux tubes within the closed-field region concentrates magnetic shear and free energy near the polarity inversion line, forming the filament channel. Onset of reconnection between this field and the external, unsheared, open field releases stored energy to drive the impulsive jet. We discuss the results of our new simulations with implications for understanding solar activity and space weather.

  11. Filament Channel Formation Via Magnetic Helicity Condensation

    NASA Astrophysics Data System (ADS)

    Knizhnik, Kalman Joshua; Antiochos, Spiro K.; DeVore, C. Richard

    2015-04-01

    A major unexplained feature of the solar atmosphere is the accumulation of magnetic shear, in the form of filament channels, at photospheric polarity inversion lines (PILs). In addition to free energy, this shear also represents magnetic helicity, which is conserved under reconnection. In this work, we address the problem of filament channel formation and show how they acquire their shear and magnetic helicity. The results of 3D simulations using the Adaptively Refined Magnetohydrodynamics Solver (ARMS) are presented that support the model of filament channel formation by magnetic helicity condensation developed by Antiochos (2013). We consider the convective twisting of a quasi-potential flux system that is bounded by a PIL and contains a coronal hole (CH). The magnetic helicity injected by the small-scale photospheric motions is shown to inverse-cascade up to the largest allowable scales that defined the closed flux system: the PIL and the CH. This process produces field lines that are both sheared and smooth, and are sheared in opposite senses at the PIL and the CH. The accumulated helicity and shear flux are shown to be in excellent quantitative agreement with the helicity-condensation model. We present a detailed analysis of the simulations, including comparisons of our analytical and numerical results, and discuss their implications for observations. Our research was supported by NASA's Earth and Space Science Fellowship (K.J.K.) and Heliophysics Supporting Research (S.K.A. and C.R.D.) programs.

  12. Role of actin filaments in fusopod formation and osteoclastogenesis.

    PubMed

    Wang, Yongqiang; Brooks, Patricia Joyce; Jang, Janet Jinyoung; Silver, Alexandra Shade; Arora, Pamma D; McCulloch, Christopher A; Glogauer, Michael

    2015-07-01

    Cell fusion process is a critical, rate-limiting step in osteoclastogenesis but the mechanisms that regulate fusopod formation are not defined. We characterized fusopod generation in cultured pre-osteoclasts derived from cells stably transfected with a plasmid that expressed a short, actin filament binding peptide (Lifeact) fused to mEGFP that enables localization of actin filaments in living cells. Fusion was initiated at fusopods, which are cell extensions of width >2 μm and that are immunostained for myosin-X at the extension tips. Fusopods formed at the leading edge of larger migrating cells and from the tail of adjacent smaller cells, both of which migrated in the same direction. Staining for DC-STAMP was circumferential and did not localize to cell-cell fusion sites. Compared with wild-type cells, monocytes null for Rac1 exhibited 6-fold fewer fusopods and formed 4-fold fewer multinucleated osteoclasts. From time-lapse images we found that fusion was temporally related to the formation of coherent and spatially isolated bands of actin filaments that originated in cell bodies and extended into the fusopods. These bands of actin filaments were involved in cell fusion after approaching cells formed initial contacts. We conclude that the formation of fusopods is regulated by Rac1 to initiate intercellular contact during osteoclastogenesis. This step is followed by the tightly regulated assembly of bands of actin filaments in fusopods, which lead to closure of the intercellular gap and finally, cell fusion. These novel, actin-dependent processes are important for fusion processes in osteoclastogenesis.

  13. Dynamics of filament formation in a Kerr medium

    SciTech Connect

    Centurion, Martin; Pu Ye; Tsang, Mankei; Psaltis, Demetri

    2005-06-15

    We have studied the large-scale beam breakup and filamentation of femtosecond pulses in a Kerr medium. We have experimentally monitored the formation of stable light filaments, conical emission, and interactions between filaments. Three major stages lead to the formation of stable light filaments: First the beam breaks up into a pattern of connected lines (constellation), then filaments form on the constellations, and finally the filaments release a fraction of their energy through conical emission. We observed a phase transition to a faster filamentation rate at the onset of conical emission. We attribute this to the interaction of conical emissions with the constellation which creates additional filaments. Numerical simulations show good agreement with the experimental results.

  14. Galactic cold cores. VIII. Filament formation and evolution: Filament properties in context with evolutionary models

    NASA Astrophysics Data System (ADS)

    Rivera-Ingraham, A.; Ristorcelli, I.; Juvela, M.; Montillaud, J.; Men'shchikov, A.; Malinen, J.; Pelkonen, V.-M.; Marston, A.; Martin, P. G.; Pagani, L.; Paladini, R.; Paradis, D.; Ysard, N.; Ward-Thompson, D.; Bernard, J.-P.; Marshall, D. J.; Montier, L.; Tóth, L. V.

    2017-05-01

    Context. The onset of star formation is intimately linked with the presence of massive unstable filamentary structures. These filaments are therefore key for theoretical models that aim to reproduce the observed characteristics of the star formation process in the Galaxy. Aims: As part of the filament study carried out by the Herschel Galactic Cold Cores Key Programme, here we study and discuss the filament properties presented in GCC VII (Paper I) in context with theoretical models of filament formation and evolution. Methods: A conservatively selected sample of filaments located at a distance D< 500 pc was extracted from the GCC fields with the getfilaments algorithm. The physical structure of the filaments was quantified according to two main components: the central (Gaussian) region of the filament (core component), and the power-law-like region dominating the filament column density profile at larger radii (wing component). The properties and behaviour of these components relative to the total linear mass density of the filament and the column density of its environment were compared with the predictions from theoretical models describing the evolution of filaments under gravity-dominated conditions. Results: The feasibility of a transition from a subcritical to supercritical state by accretion at any given time is dependent on the combined effect of filament intrinsic properties and environmental conditions. Reasonably self-gravitating (high Mline,core) filaments in dense environments (AV≳ 3 mag) can become supercritical on timescales of t 1 Myr by accreting mass at constant or decreasing width. The trend of increasing Mline,tot (Mline,core and Mline,wing) and ridge AV with background for the filament population also indicates that the precursors of star-forming filaments evolve coevally with their environment. The simultaneous increase of environment and filament AV explains the observed association between dense environments and high Mline,core values

  15. Filament formation as a scale-free process

    NASA Astrophysics Data System (ADS)

    Vazquez-Semadeni, Enrique

    2015-08-01

    I will discuss the formation of filaments in molecular clouds, and its potential similarity with filament formation in the cosmic web. First, I will recall the formation mechanism of giant molecular clouds (GMCs) and their likely state of global, hierarchical gravitational collapse, which amplifies any anisotropies of the initial configuration. I will then briefly recall the density and structure produced by this mechanism, emphasizing the fact that filaments are flow features, funneling material from the cloud to the star-forming cores, rather than static objects. I will conclude with a comparison of the physical processes operating in GMC filament formation to that operating on the formation of extragalactic filaments, to conclude with the question of the extent to which the two processes are comparable.

  16. Tropomyosin - master regulator of actin filament function in the cytoskeleton.

    PubMed

    Gunning, Peter W; Hardeman, Edna C; Lappalainen, Pekka; Mulvihill, Daniel P

    2015-08-15

    Tropomyosin (Tpm) isoforms are the master regulators of the functions of individual actin filaments in fungi and metazoans. Tpms are coiled-coil parallel dimers that form a head-to-tail polymer along the length of actin filaments. Yeast only has two Tpm isoforms, whereas mammals have over 40. Each cytoskeletal actin filament contains a homopolymer of Tpm homodimers, resulting in a filament of uniform Tpm composition along its length. Evidence for this 'master regulator' role is based on four core sets of observation. First, spatially and functionally distinct actin filaments contain different Tpm isoforms, and recent data suggest that members of the formin family of actin filament nucleators can specify which Tpm isoform is added to the growing actin filament. Second, Tpms regulate whole-organism physiology in terms of morphogenesis, cell proliferation, vesicle trafficking, biomechanics, glucose metabolism and organ size in an isoform-specific manner. Third, Tpms achieve these functional outputs by regulating the interaction of actin filaments with myosin motors and actin-binding proteins in an isoform-specific manner. Last, the assembly of complex structures, such as stress fibers and podosomes involves the collaboration of multiple types of actin filament specified by their Tpm composition. This allows the cell to specify actin filament function in time and space by simply specifying their Tpm isoform composition.

  17. Cysteine-rich protein 2 accelerates actin filament cluster formation

    PubMed Central

    Shinohara, Satoko; Takaoka, Shunpei; Miyake, Jun

    2017-01-01

    Filamentous actin (F-actin) forms many types of structures and dynamically regulates cell morphology and movement, and plays a mechanosensory role for extracellular stimuli. In this study, we determined that the smooth muscle-related transcription factor, cysteine-rich protein 2 (CRP2), regulates the supramolecular networks of F-actin. The structures of CRP2 and F-actin in solution were analyzed by small-angle X-ray solution scattering (SAXS). The general shape of CRP2 was partially unfolded and relatively ellipsoidal in structure, and the apparent cross sectional radius of gyration (Rc) was about 15.8 Å. The predicted shape, derived by ab initio modeling, consisted of roughly four tandem clusters: LIM domains were likely at both ends with the middle clusters being an unfolded linker region. From the SAXS analysis, the Rc of F-actin was about 26.7 Å, and it was independent of CRP2 addition. On the other hand, in the low angle region of the CRP2-bound F-actin scattering, the intensities showed upward curvature with the addition of CRP2, which indicates increasing branching of F-actin following CRP2 binding. From biochemical analysis, the actin filaments were augmented and clustered by the addition of CRP2. This F-actin clustering activity of CRP2 was cooperative with α-actinin. Thus, binding of CRP2 to F-actin accelerates actin polymerization and F-actin cluster formation. PMID:28813482

  18. Formation of interstellar filaments: the role of magnetic fields

    NASA Astrophysics Data System (ADS)

    Ntormousi, Evangelia; Hennebelle, Patrick

    2014-07-01

    The filamentary structure of interstellar matter and its potential link to star formation has been brought back into focus recently by high resolution observational surveys. The densest of these filaments host pre-stellar and star forming cores, so explaining their properties is tightly correlated to revealing the initial conditions for star formation. To that end, in this work we employ high-resolution, 3D MHD simulations performed with the AMR code RAMSES to investigate two filament formation mechanisms: turbulence and sheet fragmentation. The first series of simulations has as a particular aim to address the origin of the characteristic filament thickness found in observations. Starting from the hypothesis that diffusive processes are responsible, our numerical experiments consist of (driven or decaying) ideal and non-ideal MHD turbulence, at a resolution that greatly exceeds the reported 0.1pc thickness. The comparison points to ion-neutral friction as an excellent candidate for setting a characteristic scale. In this picture dense filaments are the diffusive end of the turbulent cascade, an interpretation with important implications for our understanding of the dynamical behavior of the ISM. A second series of simulations investigates filament formation by the fragmentation of supershells, a scenario inspired by the analytical work of Nagai (1998). We find a striking difference between hydrodynamical and MHD runs as in the first case the sheets fragment into small cores, while in the latter they produce large filaments. In addition though, we see that low-density filaments preferentially form along the dominant component of the magnetic field. In this scenario filaments are prominent features in the ISM, but their fate is still determined by the local magnetic field. A detailed comparison of the filament properties between the two runs is work in progress and will reveal the physical mechanisms responsible for shaping the ISM and setting the initial conditions

  19. Biophysics of filament length regulation by molecular motors

    PubMed Central

    Kuan, Hui-Shun; Betterton, M. D.

    2013-01-01

    Regulating physical size is an essential problem that biological organisms must solve from the subcellular to the organismal scales, but it is not well understood what physical principles and mechanisms organisms use to sense and regulate their size. Any biophysical size-regulation scheme operates in a noisy environment and must be robust to other cellular dynamics and fluctuations. This work develops theory of filament length regulation inspired by recent experiments on kinesin-8 motor proteins, which move with directional bias on microtubule filaments and alter microtubule dynamics. Purified kinesin-8 motors can depolymerize chemically-stabilized microtubules. In the length-dependent depolymerization model, the rate of depolymerization tends to increase with filament length, because long filaments accumulate more motors at their tips and therefore shorten more quickly. When balanced with a constant filament growth rate, this mechanism can lead to a fixed polymer length. However, the mechanism by which kinesin-8 motors affect the length of dynamic microtubules in cells is less clear. We study the more biologically realistic problem of microtubule dynamic instability modulated by a motor-dependent increase in the filament catastrophe frequency. This leads to a significant decrease in the mean filament length and a narrowing of the filament length distribution. The results improve our understanding of the biophysics of length regulation in cells. PMID:23587993

  20. Biophysics of filament length regulation by molecular motors.

    PubMed

    Kuan, Hui-Shun; Betterton, M D

    2013-06-01

    Regulating physical size is an essential problem that biological organisms must solve from the subcellular to the organismal scales, but it is not well understood what physical principles and mechanisms organisms use to sense and regulate their size. Any biophysical size-regulation scheme operates in a noisy environment and must be robust to other cellular dynamics and fluctuations. This work develops theory of filament length regulation inspired by recent experiments on kinesin-8 motor proteins, which move with directional bias on microtubule filaments and alter microtubule dynamics. Purified kinesin-8 motors can depolymerize chemically-stabilized microtubules. In the length-dependent depolymerization model, the rate of depolymerization tends to increase with filament length, because long filaments accumulate more motors at their tips and therefore shorten more quickly. When balanced with a constant filament growth rate, this mechanism can lead to a fixed polymer length. However, the mechanism by which kinesin-8 motors affect the length of dynamic microtubules in cells is less clear. We study the more biologically realistic problem of microtubule dynamic instability modulated by a motor-dependent increase in the filament catastrophe frequency. This leads to a significant decrease in the mean filament length and a narrowing of the filament length distribution. The results improve our understanding of the biophysics of length regulation in cells.

  1. Investigating Filamentous Growth and Biofilm/Mat Formation in Budding Yeast

    PubMed Central

    Cullen, Paul J.

    2015-01-01

    In response to nutrient limitation, budding yeast can undergo filamentous growth by differentiating into elongated chains of interconnected cells. Filamentous growth is regulated by signal transduction pathways that oversee the reorganization of cell polarity, changes to the cell cycle, and an increase in cell adhesion that occur in response to nutrient limitation. Each of these changes can be easily measured. Yeast can also grow colonially atop surfaces in a biofilm or mat of connected cells. Filamentous growth and biofilm/mat formation require cooperation among individuals; therefore, studying these responses can shed light on the origin and genetic basis of multicellular behaviors. The assays introduced here can be used to study analogous behaviors in fungal pathogens, which require filamentous growth and biofilm/mat formation for virulence. PMID:25734073

  2. Reorganization of actin filaments by ADF/cofilin is involved in formation of microtubule structures during Xenopus oocyte maturation

    PubMed Central

    Yamagishi, Yuka; Abe, Hiroshi

    2015-01-01

    We examined the reorganization of actin filaments and microtubules during Xenopus oocyte maturation. Surrounding the germinal vesicle (GV) in immature oocytes, the cytoplasmic actin filaments reorganized to accumulate beneath the vegetal side of the GV, where the microtubule-organizing center and transient microtubule array (MTOC-TMA) assembled, just before GV breakdown (GVBD). Immediately after GVBD, both Xenopus ADF/cofilin (XAC) and its phosphatase Slingshot (XSSH) accumulated into the nuclei and intranuclear actin filaments disassembled from the vegetal side with the shrinkage of the GV. As the MTOC-TMA developed well, cytoplasmic actin filaments were retained at the MTOC-TMA base region. Suppression of XAC dephosphorylation by anti-XSSH antibody injection inhibited both actin filament reorganization and proper formation and localization of both the MTOC-TMA and meiotic spindles. Stabilization of actin filaments by phalloidin also inhibited formation of the MTOC-TMA and disassembly of intranuclear actin filaments without affecting nuclear shrinkage. Nocodazole also caused the MTOC-TMA and the cytoplasmic actin filaments at its base region to disappear, which further impeded disassembly of intranuclear actin filaments from the vegetal side. XAC appears to reorganize cytoplasmic actin filaments required for precise assembly of the MTOC and, together with the MTOC-TMA, regulate the intranuclear actin filament disassembly essential for meiotic spindle formation. PMID:26424802

  3. Filament fragmentation in high-mass star formation

    NASA Astrophysics Data System (ADS)

    Beuther, H.; Ragan, S. E.; Johnston, K.; Henning, Th.; Hacar, A.; Kainulainen, J. T.

    2015-12-01

    Context. Filamentary structures in the interstellar medium are crucial ingredients of the star formation process. They fragment to form individual star-forming cores, and at the same time they may also funnel gas toward the central gas cores, providing an additional gas reservoir. Aims: We want to resolve the length scales for filament formation and fragmentation (resolution ≤0.1 pc), in particular the Jeans length and cylinder fragmentation scale. Methods: We have observed the prototypical high-mass star-forming filament IRDC 18223 with the Plateau de Bure Interferometer (PdBI) in the 3.2 mm continuum and N2H+(1-0) line emission in a ten-field mosaic at a spatial resolution of ~ 4'' (~14 000 au). Results: The dust continuum emission resolves the filament into a chain of at least 12 relatively regularly spaced cores. The mean separation between cores is ~0.40(± 0.18) pc. While this is approximately consistent with the fragmentation of an infinite, isothermal, and gravitationally bound gas cylinder, a high mass-to-length ratio of M/l ≈ 1000 M⊙ pc-1 requires additional turbulent and/or magnetic support against radial collapse of the filament. The N2H+(1-0) data reveal a velocity gradient perpendicular to the main filament. Although rotation of the filament cannot be excluded, the data are also consistent with the main filament being comprised of several velocity-coherent subfilaments. Furthermore, this velocity gradient perpendicular to the filament resembles results toward Serpens south that are interpreted as signatures of filament formation within magnetized and turbulent sheet-like structures. Lower-density gas tracers ([CI] and C18O) reveal a similar red- and blueshifted velocity structure on scales around 60'' east and west of the filament. This may tentatively be interpreted as a signature of the large-scale cloud and the smaller scale filament being kinematically coupled. We do not identify a velocity gradient along the axis of the filament. This may

  4. Triggered star formation in the inner filament of Centaurus A

    NASA Astrophysics Data System (ADS)

    Crockett, R. Mark; Shabala, Stanislav S.; Kaviraj, Sugata; Antonuccio-Delogu, Vincenzo; Silk, Joseph; Mutchler, Max; O'Connell, Robert W.; Rejkuba, Marina; Whitmore, Bradley C.; Windhorst, Rogier A.

    2012-04-01

    We present recent Hubble Space Telescope observations of the inner filament of Centaurus A, using the new Wide Field Camera 3 (WFC3) F225W, F657N and F814W filters. We find a young stellar population near the south-west tip of the filament. Combining the WFC3 data set with archival Advanced Camera for Surveys (ACS) F606W observations, we are able to constrain the ages of these stars to ≲10 Myr, with best-fitting ages of 1-4 Myr. No further recent star formation is found along the filament. Based on the location and age of this stellar population, and the fact that there is no radio lobe or jet activity near the star formation, we propose an updated explanation for the origin of the inner filament. Sutherland et al. suggested that radio jet-induced shocks can drive the observed optical line emission. We argue that such shocks can naturally arise due to a weak cocoon-driven bow shock (rather than from the radio jet directly), propagating through the diffuse interstellar medium from a location near the inner northern radio lobe. The shock can overrun a molecular cloud, triggering star formation in the dense molecular cores. Ablation and shock heating of the diffuse gas then give rise to the observed optical line and X-ray emission. Deeper X-ray observations should show more diffuse emission along the filament.

  5. Simulations of Filament Channel Formation in a Coronal Magnetic Field

    NASA Astrophysics Data System (ADS)

    Knizhnik, Kalman; DeVore, C. Richard; Antiochos, Spiro K.

    2016-05-01

    A major unanswered problem in solar physics has been explaining the presence of sheared filament channels above photospheric polarity inversion lines (PILs) and the simultaneous lack of structure in the ‘loop’ portion of the coronal magnetic field. The shear inherent in filament channels represents not only a form of magnetic energy, but also magnetic helicity. As a result, models of filament channel formation need to explain not only why helicity is observed above PILs, but also why it is apparently not observed anywhere else in the corona. Previous results (Knizhnik, Antiochos & DeVore, 2015) have suggested that any helicity injected into the coronal field inverse-cascades in scale, a process known as magnetic helicity condensation (Antiochos, 2013). In this work, we present high resolution numerical simulations of photospheric helicity injection into a coronal magnetic field that contains both a PIL and a coronal hole (CH). We show conclusively that the inverse cascade of magnetic helicity terminates at the PIL, resulting in the formation of highly sheared filament channels and a smooth, untwisted corona. We demonstrate that even though magnetic helicity is injected throughout the flux system, it accumulates only at the PIL, where it manifests itself in the form of highly sheared filament channels, while any helicity obtained by the CH is ejected out of the system. We show that the formation of filament channels is both qualitatively and quantitatively in agreement with observations and discuss the implications of our simulations for observations.This work was supported by the NASA Earth and Space Science Fellowship, LWS TR&T and H-SR Programs.

  6. Rapid Formation and Disappearance of a Filament Barb

    NASA Astrophysics Data System (ADS)

    Joshi, Anand D.; Srivastava, Nandita; Mathew, Shibu K.; Martin, Sara F.

    2013-11-01

    We present observations of an activated quiescent filament obtained in Hα from the high-resolution Dutch Open Telescope (DOT) on 20 August 2010. The filament developed a barb in 10 min, which disappeared within the next 35 min. A data set from the DOT spanning 2 h was used to analyse this event. Line-of-sight velocity maps were constructed from the Doppler images, which reveal flows in filament spine during this period. Photospheric magnetograms were used from the Helioseismic and Magnetic Imager (HMI) on board the Solar Dynamics Observatory (SDO) to determine the changes in magnetic flux in the region surrounding the barb location. The analysis shows flows in the filament spine towards the barb location preceding its formation, and flows in the barb towards the spine during its disappearance. Magnetograms reveal patches of minority polarity flux close to the end of the barb at its greatest elongation. The flows in the spine and barbs are along numerous threads that compose these typical filament structures. The flows are consistent with field-aligned threads and demonstrate that the replacement time of the mass in barbs, and by inference, in the spine is very rapid.

  7. Dynamic Star Formation in the Massive DR21 Filament

    SciTech Connect

    Schneider, N.; Csengeri, T.; Bontemps, S.; Motte, F.; Simon, R.; Hennebelle, P.; Federrath, C.; Klessen, R.; /ZAH, Heidelberg /KIPAC, Menlo Park

    2010-08-25

    The formation of massive stars is a highly complex process in which it is unclear whether the star-forming gas is in global gravitational collapse or an equilibrium state supported by turbulence and/or magnetic fields. By studying one of the most massive and dense star-forming regions in the Galaxy at a distance of less than 3 kpc, i.e. the filament containing the well-known sources DR21 and DR21(OH), we attempt to obtain observational evidence to help us to discriminate between these two views. We use molecular line data from our {sup 13}CO 1 {yields} 0, CS 2 {yields} 1, and N{sub 2}H{sup +} 1 {yields} 0 survey of the Cygnus X region obtained with the FCRAO and CO, CS, HCO{sup +}, N{sub 2}H{sup +}, and H{sub 2}CO data obtained with the IRAM 30m telescope. We observe a complex velocity field and velocity dispersion in the DR21 filament in which regions of the highest column-density, i.e., dense cores, have a lower velocity dispersion than the surrounding gas and velocity gradients that are not (only) due to rotation. Infall signatures in optically thick line profiles of HCO{sup +} and {sup 12}CO are observed along and across the whole DR21 filament. By modelling the observed spectra, we obtain a typical infall speed of {approx}0.6 km s{sup -1} and mass accretion rates of the order of a few 10{sup -3} M{sub {circle_dot}} yr{sup -1} for the two main clumps constituting the filament. These massive clumps (4900 and 3300 M{sub {circle_dot}} at densities of around 10{sup 5} cm{sup -3} within 1 pc diameter) are both gravitationally contracting. The more massive of the clumps, DR21(OH), is connected to a sub-filament, apparently 'falling' onto the clump. This filament runs parallel to the magnetic field. Conclusions. All observed kinematic features in the DR21 filament (velocity field, velocity dispersion, and infall), its filamentary morphology, and the existence of (a) sub-filament(s) can be explained if the DR21 filament was formed by the convergence of flows on large

  8. FILAMENT FORMATION BY ESCHERICHIA COLI AT INCREASED HYDROSTATIC PRESSURES1

    PubMed Central

    Zobell, Claude E.; Cobet, Andre B.

    1964-01-01

    ZoBell, Claude E. (University of California, La Jolla), and Andre B. Cobet. Filament formation by Escherichia coli at increased hydrostatic pressures. J. Bacteriol. 87:710–719. 1964.—The reproduction as well as the growth of Escherichia coli is retarded by hydrostatic pressures ranging from 200 to 500 atm. Reproduction was indicated by an increase in the number of cells determined by plating on EMB Agar as well as by direct microscopic counts. Growth, which is not necessarily synonymous with reproduction, was indicated by increase in dry weight and protein content of the bacterial biomass. At increased pressures, cells of three different strains of E. coli tended to form long filaments. Whereas most normal cells of E. coli that developed at 1 atm were only about 2 μ long, the mean length of those that developed at 475 atm was 2.93 μ for strain R4, 3.99 μ for strain S, and 5.82 μ for strain B cells. Nearly 90% of the bacterial biomass produced at 475 atm by strain B was found in filaments exceeding 5 μ in length; 74.7 and 16.4% of the biomass produced at 475 atm by strains S and R4, respectively, occurred in such filaments. Strain R4 formed fewer and shorter (5 to 35 μ) filaments than did the other two strains, whose filaments ranged in length from 5 to >100 μ. The bacterial biomass produced at all pressures had approximately the same content of protein and nucleic acids. But at increased pressures appreciably more ribonucleic acid (RNA) and proportionately less deoxyribonucleic acid (DNA) was found per unit of biomass. Whereas the RNA content per cell increased with cell length, the amount of DNA was nearly the same in long filaments formed at increased pressure as in cells of normal length formed at 1 atm. The inverse relationship between the concentration of DNA and cell length in all three strains of E. coli suggests that the failure of DNA to replicate at increased pressure may be responsible for a repression of cell division and consequent filament

  9. Induction of Filament Formation and Thymineless Death in Escherichia coli K-12

    PubMed Central

    Gherardi, M.; Sicard, N.

    1970-01-01

    The study of isogenic strains of Escherichia coli K-12 carrying mutations which control filament formation after ultraviolet irradiation showed that there is not necessarily a relationship between filament formation and thymineless death. PMID:4908682

  10. Ack kinase regulates CTP synthase filaments during Drosophila oogenesis.

    PubMed

    Strochlic, Todd I; Stavrides, Kevin P; Thomas, Sam V; Nicolas, Emmanuelle; O'Reilly, Alana M; Peterson, Jeffrey R

    2014-11-01

    The enzyme CTP synthase (CTPS) dynamically assembles into macromolecular filaments in bacteria, yeast, Drosophila, and mammalian cells, but the role of this morphological reorganization in regulating CTPS activity is controversial. During Drosophila oogenesis, CTPS filaments are transiently apparent in ovarian germline cells during a period of intense genomic endoreplication and stockpiling of ribosomal RNA. Here, we demonstrate that CTPS filaments are catalytically active and that their assembly is regulated by the non-receptor tyrosine kinase DAck, the Drosophila homologue of mammalian Ack1 (activated cdc42-associated kinase 1), which we find also localizes to CTPS filaments. Egg chambers from flies deficient in DAck or lacking DAck catalytic activity exhibit disrupted CTPS filament architecture and morphological defects that correlate with reduced fertility. Furthermore, ovaries from these flies exhibit reduced levels of total RNA, suggesting that DAck may regulate CTP synthase activity. These findings highlight an unexpected function for DAck and provide insight into a novel pathway for the developmental control of an essential metabolic pathway governing nucleotide biosynthesis.

  11. Cluster Formation Triggered by Filament Collisions in Serpens South

    NASA Astrophysics Data System (ADS)

    Nakamura, Fumitaka

    2015-08-01

    The Serpens South infrared darkcloud consists of several filamentary ridges, some of which fragment into dense clumps. On the basis of CCS, N2H+, SiO observations, we investigated the kinematics and chemical evolution of these filamentary ridges. We find that CCS is extremely strong along the main filament, in particular toward northern precluster clump. We emphasize that Serpens South is the first protocluster clump where strong CCS emission is detected. We identified several ridges with different velocity. These ridges appear to converge toward the protocluster clump, suggesting that the collisions of these ridges may have trigerred cluster formation. The collisions presumably happened within the last half Myr because the chemical evolution calculations indicate that CCS is abundant only in the first half Myr. We also speculate that these filamentary ridges might be formed in a compressed turbulent region created by a larger-scale cloud-cloud collision and subsequent filament collision triggered cluster formation.

  12. Scaling During Drop Formation and Filament (Thread) Breakup

    NASA Astrophysics Data System (ADS)

    Wagoner, Brayden; Thete, Sumeet; Basaran, Osman

    2016-11-01

    Many free surface flows such as drop formation, filament (thread) breakup, and drop coalescence are important in applications as diverse as ink jet printing, atomization, and emulsion science and technology. A common feature of these flows is that they all exhibit finite time singularities. When a liquid filament undergoes capillary thinning and tends toward pinch-off, it is instructive to monitor how certain quantities, such as the thread's radius, vary with time remaining until the pinch-off singularity. Experimental determination of this so-called scaling behavior of thread radius and other quantities is important for testing scaling theories and the accuracy of numerical simulations of free surface flows. Conversely, the experimental measurements can be used to develop new theories when none are available. In this talk, we will present some novel ways of experimentally measuring scaling behaviors. The results will be highlighted in terms of experiments involving the formation and breakup of drops and filaments of (a) simple or pure Newtonian fluids and also (b) particle-laden liquids or suspensions containing non-Brownian particles.

  13. Magnetohydrodynamics of Wind-Cloud Interactions: Filament Formation in the Interstellar Medium

    NASA Astrophysics Data System (ADS)

    Banda-Barragán, Wladimir E.

    2016-08-01

    Filaments are ubiquitous in the interstellar medium, yet their formation, internal structure, magnetic properties, and longevity have not been analysed in detail. In this thesis I report the results from a comprehensive numerical study that investigates the characteristics, formation, dynamics, and global evolution of filamentary structures arising from (magneto)hydrodynamic interactions between supersonic winds and interstellar clouds. Here I improve on previous wind-cloud simulations by utilising higher numerical resolutions, sharper density contrasts, more complex magnetic field configurations, and more realistic systems with turbulent clouds. I use gas multi-tracking algorithms and state-of-the-art visualisation techniques to study the physical mechanisms acting upon wind-swept clouds. I find that material originally in the envelopes of the clouds is removed and transported downstream to form filamentary tails, while the cores of the clouds serve as footpoints and late-stage outer layers of these low-density tails. The evolution of filaments comprises four phases: 1) tail formation, 2) tail erosion, 3) footpoint dispersion, and 4) filament free floating. Overall, wind-cloud interactions produce filaments with aspect ratios >10, lateral expansions 1-3 of the core radius, mixing fractions 10-30%, velocity dispersions 0.02-0.05 of the wind speed, and magnetic field amplifications by factors of 10-100. I find that the strength of magnetic fields regulates vorticity production: sinuous filamentary towers arise in non-magnetic environments, while strong magnetic fields inhibit small-scale Kelvin-Helmholtz perturbations at boundary layers making tails less turbulent. The orientation of magnetic fields also influences the morphology of filaments: magnetic field components aligned with the direction of the wind favour the formation of pressure-confined flux ropes inside the tails, whilst transverse components tend to form current sheets and favour the growth of Rayleigh

  14. Mechanical force antagonizes the inhibitory effects of RecX on RecA filament formation in Mycobacterium tuberculosis.

    PubMed

    Le, Shimin; Chen, Hu; Zhang, Xinghua; Chen, Jin; Patil, K Neelakanteshwar; Muniyappa, Kalappa; Yan, Jie

    2014-10-29

    Efficient bacterial recombinational DNA repair involves rapid cycles of RecA filament assembly and disassembly. The RecX protein plays a crucial inhibitory role in RecA filament formation and stability. As the broken ends of DNA are tethered during homologous search, RecA filaments assembled at the ends are likely subject to force. In this work, we investigated the interplay between RecX and force on RecA filament formation and stability. Using magnetic tweezers, at single molecular level, we found that Mycobacterium tuberculosis (Mt) RecX could catalyze stepwise de-polymerization of preformed MtRecA filament in the presence of ATP hydrolysis at low forces (<7 pN). However, applying larger forces antagonized the inhibitory effects of MtRecX, and a partially de-polymerized MtRecA filament could re-polymerize in the presence of MtRecX, which cannot be explained by previous models. Theoretical analysis of force-dependent conformational free energies of naked ssDNA and RecA nucleoprotein filament suggests that mechanical force stabilizes RecA filament, which provides a possible mechanism for the observation. As the antagonizing effect of force on the inhibitory function of RecX takes place in a physiological range; these findings broadly suggest a potential mechanosensitive regulation during homologous recombination.

  15. Regulation of Sodium Channel Activity by Capping of Actin Filaments

    PubMed Central

    Shumilina, Ekaterina V.; Negulyaev, Yuri A.; Morachevskaya, Elena A.; Hinssen, Horst; Khaitlina, Sofia Yu

    2003-01-01

    Ion transport in various tissues can be regulated by the cortical actin cytoskeleton. Specifically, involvement of actin dynamics in the regulation of nonvoltage-gated sodium channels has been shown. Herein, inside-out patch clamp experiments were performed to study the effect of the heterodimeric actin capping protein CapZ on sodium channel regulation in leukemia K562 cells. The channels were activated by cytochalasin-induced disruption of actin filaments and inactivated by G-actin under ionic conditions promoting rapid actin polymerization. CapZ had no direct effect on channel activity. However, being added together with G-actin, CapZ prevented actin-induced channel inactivation, and this effect occurred at CapZ/actin molar ratios from 1:5 to 1:100. When actin was allowed to polymerize at the plasma membrane to induce partial channel inactivation, subsequent addition of CapZ restored the channel activity. These results can be explained by CapZ-induced inhibition of further assembly of actin filaments at the plasma membrane due to the modification of actin dynamics by CapZ. No effect on the channel activity was observed in response to F-actin, confirming that the mechanism of channel inactivation does not involve interaction of the channel with preformed filaments. Our data show that actin-capping protein can participate in the cytoskeleton-associated regulation of sodium transport in nonexcitable cells. PMID:12686620

  16. Transcriptional regulation of plant cell wall degradation by filamentous fungi.

    PubMed

    Aro, Nina; Pakula, Tiina; Penttilä, Merja

    2005-09-01

    Plant cell wall consists mainly of the large biopolymers cellulose, hemicellulose, lignin and pectin. These biopolymers are degraded by many microorganisms, in particular filamentous fungi, with the aid of extracellular enzymes. Filamentous fungi have a key role in degradation of the most abundant biopolymers found in nature, cellulose and hemicelluloses, and therefore are essential for the maintenance of the global carbon cycle. The production of plant cell wall degrading enzymes, cellulases, hemicellulases, ligninases and pectinases, is regulated mainly at the transcriptional level in filamentous fungi. The genes are induced in the presence of the polymers or molecules derived from the polymers and repressed under growth conditions where the production of these enzymes is not necessary, such as on glucose. The expression of the genes encoding the enzymes is regulated by various environmental and cellular factors, some of which are common while others are more unique to either a certain fungus or a class of enzymes. This review summarises our current knowledge on the transcriptional regulation, focusing on the recently characterized transcription factors that regulate genes coding for enzymes involved in the breakdown of plant cell wall biopolymers.

  17. GRAVITATIONAL COLLAPSE AND FILAMENT FORMATION: COMPARISON WITH THE PIPE NEBULA

    SciTech Connect

    Heitsch, Fabian; Ballesteros-Paredes, Javier; Hartmann, Lee

    2009-10-20

    Recent models of molecular cloud formation and evolution suggest that such clouds are dynamic and generally exhibit gravitational collapse. We present a simple analytic model of global collapse onto a filament and compare this with our numerical simulations of the flow-driven formation of an isolated molecular cloud to illustrate the supersonic motions and infall ram pressures expected in models of gravity-driven cloud evolution. We compare our results with observations of the Pipe Nebula, an especially suitable object for our purposes as its low star formation activity implies insignificant perturbations from stellar feedback. We show that our collapsing cloud model can explain the magnitude of the velocity dispersions seen in the {sup 13}CO filamentary structure by Onishi et al. and the ram pressures required by Lada et al. to confine the lower-mass cores in the Pipe Nebula. We further conjecture that higher-resolution simulations will show small velocity dispersions in the densest core gas, as observed, but which are infall motions and not supporting turbulence. Our results point out the inevitability of ram pressures as boundary conditions for molecular cloud filaments, and the possibility that especially lower-mass cores still can be accreting mass at significant rates, as suggested by observations.

  18. The filamentous fungal pellet and forces driving its formation.

    PubMed

    Zhang, Jianguo; Zhang, Jining

    2016-12-01

    Filamentous fungi play an important role not only in the bio-manufacturing of value-added products, but also in bioenergy and environmental research. The bioprocess manipulation of filamentous fungi is more difficult than that of other microbial species because of their different pellet morphologies and the presence of tangled mycelia under different cultivation conditions. Fungal pellets, which have the advantages of harvest ease, low fermentation broth viscosity and high yield of some proteins, have been used for a long time. Many attempts have been made to establish the relationship between pellet and product yield using quantitative approaches. Fungal pellet formation is attributed to the combination of electrostatic interactions, hydrophobicity and specific interactions from spore wall components. Electrostatic interactions result from van der Waals forces and negative charge repulsion from carboxyl groups in the spore wall structure. Electrostatic interactions are also affected by counter-ions (cations) and the physiologic conditions of spores that modify the carboxyl groups. Fungal aggregates are promoted by the hydrophobicity generated by hydrophobins, which form a hydrophobic coat that covers the spore. The specific interactions of spore wall components contribute to spore aggregation through salt bridging. A model of spore aggregation was proposed based on these forces. Additionally, some challenges were addressed, including the limitations of research techniques, the quantitative determination of forces and the complex information of biological systems, to clarify the mechanism of fungal pellet formation.

  19. Dynamics of the current filament formation and its steady-state characteristics in chalcogenide based PCM

    NASA Astrophysics Data System (ADS)

    Bogoslovskiy, Nikita; Tsendin, Konstantin

    2017-03-01

    In the phase-change memory (PCM) crystallization occurs in the high-current filament which forms during switching to the conductive state. In the present paper we conduct a numerical modeling of the current filament formation dynamics in thin chalcogenide films using an electronic-thermal model based on negative-U centers tunnel ionization and Joule heating. The key role of inhomogeneities in the filament formation process is shown. Steady-state filament parameters were obtained from the analysis of the stationary heat conduction equation. The filament formation dynamics and the steady-state filament radius and temperature could be controlled by material parameters and contact resistance. Consequently it is possible to control the size of the region wherein crystallization occurs. A good agreement with numerous experimental data leads to the conclusion that thermal effects play a significant role in CGS conduction and high-current filament formation while switching.

  20. Cluster Formation Triggered by Filament Collisions in Serpens South

    NASA Astrophysics Data System (ADS)

    Nakamura, Fumitaka; Sugitani, Koji; Tanaka, Tomohiro; Nishitani, Hiroyuki; Dobashi, Kazuhito; Shimoikura, Tomomi; Shimajiri, Yoshito; Kawabe, Ryohei; Yonekura, Yoshinori; Mizuno, Izumi; Kimura, Kimihiko; Tokuda, Kazuki; Kozu, Minato; Okada, Nozomi; Hasegawa, Yutaka; Ogawa, Hideo; Kameno, Seiji; Shinnaga, Hiroko; Momose, Munetake; Nakajima, Taku; Onishi, Toshikazu; Maezawa, Hiroyuki; Hirota, Tomoya; Takano, Shuro; Iono, Daisuke; Kuno, Nario; Yamamoto, Satoshi

    2014-08-01

    The Serpens South infrared dark cloud consists of several filamentary ridges, some of which fragment into dense clumps. On the basis of CCS (JN = 43-32), HC3N (J = 5-4), N2H+ (J = 1-0), and SiO (J = 2-1, v = 0) observations, we investigated the kinematics and chemical evolution of these filamentary ridges. We find that CCS is extremely abundant along the main filament in the protocluster clump. We emphasize that Serpens South is the first cluster-forming region where extremely strong CCS emission is detected. The CCS-to-N2H+ abundance ratio is estimated to be about 0.5 toward the protocluster clump, whereas it is about 3 in the other parts of the main filament. We identify six dense ridges with different V LSR. These ridges appear to converge toward the protocluster clump, suggesting that the collisions of these ridges may have triggered cluster formation. The collisions presumably happened within a few × 105 yr because CCS is abundant only for a short time. The short lifetime agrees with the fact that the number fraction of Class I objects, whose typical lifetime is 0.4 × 105 yr, is extremely high, about 70% in the protocluster clump. In the northern part, two ridges appear to have partially collided, forming a V-shape clump. In addition, we detected strong bipolar SiO emission that is due to the molecular outflow blowing out of the protostellar clump, as well as extended weak SiO emission that may originate from the filament collisions.

  1. CLUSTER FORMATION TRIGGERED BY FILAMENT COLLISIONS IN SERPENS SOUTH

    SciTech Connect

    Nakamura, Fumitaka; Kawabe, Ryohei; Shinnaga, Hiroko; Sugitani, Koji; Tanaka, Tomohiro; Kimura, Kimihiko; Tokuda, Kazuki; Kozu, Minato; Okada, Nozomi; Hasegawa, Yutaka; Ogawa, Hideo; Nishitani, Hiroyuki; Mizuno, Izumi; Dobashi, Kazuhito; Shimoikura, Tomomi; Shimajiri, Yoshito; Kameno, Seiji; Momose, Munetake; Nakajima, Taku; and others

    2014-08-20

    The Serpens South infrared dark cloud consists of several filamentary ridges, some of which fragment into dense clumps. On the basis of CCS (J{sub N} = 4{sub 3}-3{sub 2}), HC{sub 3}N (J = 5-4), N{sub 2}H{sup +} (J = 1-0), and SiO (J = 2-1, v = 0) observations, we investigated the kinematics and chemical evolution of these filamentary ridges. We find that CCS is extremely abundant along the main filament in the protocluster clump. We emphasize that Serpens South is the first cluster-forming region where extremely strong CCS emission is detected. The CCS-to-N{sub 2}H{sup +} abundance ratio is estimated to be about 0.5 toward the protocluster clump, whereas it is about 3 in the other parts of the main filament. We identify six dense ridges with different V {sub LSR}. These ridges appear to converge toward the protocluster clump, suggesting that the collisions of these ridges may have triggered cluster formation. The collisions presumably happened within a few × 10{sup 5} yr because CCS is abundant only for a short time. The short lifetime agrees with the fact that the number fraction of Class I objects, whose typical lifetime is 0.4 × 10{sup 5} yr, is extremely high, about 70% in the protocluster clump. In the northern part, two ridges appear to have partially collided, forming a V-shape clump. In addition, we detected strong bipolar SiO emission that is due to the molecular outflow blowing out of the protostellar clump, as well as extended weak SiO emission that may originate from the filament collisions.

  2. Formation and maintenance of nitrogen-fixing cell patterns in filamentous cyanobacteria

    PubMed Central

    Muñoz-García, Javier

    2016-01-01

    Cyanobacteria forming one-dimensional filaments are paradigmatic model organisms of the transition between unicellular and multicellular living forms. Under nitrogen-limiting conditions, in filaments of the genus Anabaena, some cells differentiate into heterocysts, which lose the possibility to divide but are able to fix environmental nitrogen for the colony. These heterocysts form a quasiregular pattern in the filament, representing a prototype of patterning and morphogenesis in prokaryotes. Recent years have seen advances in the identification of the molecular mechanism regulating this pattern. We use these data to build a theory on heterocyst pattern formation, for which both genetic regulation and the effects of cell division and filament growth are key components. The theory is based on the interplay of three generic mechanisms: local autoactivation, early long-range inhibition, and late long-range inhibition. These mechanisms can be identified with the dynamics of hetR, patS, and hetN expression. Our theory reproduces quantitatively the experimental dynamics of pattern formation and maintenance for wild type and mutants. We find that hetN alone is not enough to play the role as the late inhibitory mechanism: a second mechanism, hypothetically the products of nitrogen fixation supplied by heterocysts, must also play a role in late long-range inhibition. The preponderance of even intervals between heterocysts arises naturally as a result of the interplay between the timescales of genetic regulation and cell division. We also find that a purely stochastic initiation of the pattern, without a two-stage process, is enough to reproduce experimental observations. PMID:27162328

  3. Cardiac thin filament regulation and the Frank-Starling mechanism.

    PubMed

    Kobirumaki-Shimozawa, Fuyu; Inoue, Takahiro; Shintani, Seine A; Oyama, Kotaro; Terui, Takako; Minamisawa, Susumu; Ishiwata, Shin'ichi; Fukuda, Norio

    2014-07-01

    The heart has an intrinsic ability to increase systolic force in response to a rise in ventricular filling (the Frank-Starling law of the heart). It is widely accepted that the length dependence of myocardial activation underlies the Frank-Starling law of the heart. Recent advances in muscle physiology have enabled the identification of the factors involved in length-dependent activation, viz., titin (connectin)-based interfilament lattice spacing reduction and thin filament "on-off" regulation, with the former triggering length-dependent activation and the latter determining the number of myosin molecules recruited to thin filaments. Patients with a failing heart have demonstrated reduced exercise tolerance at least in part via depression of the Frank-Starling mechanism. Recent studies revealed that various mutations occur in the thin filament regulatory proteins, such as troponin, in the ventricular muscle of failing hearts, which consequently alter the Frank-Starling mechanism. In this article, we review the molecular mechanisms of length-dependent activation, and the influence of troponin mutations on the phenomenon.

  4. The nucleoid-associated protein Dan organizes chromosomal DNA through rigid nucleoprotein filament formation in E. coli during anoxia

    PubMed Central

    Lim, Ci Ji; Lee, Sin Yi; Teramoto, Jun; Ishihama, Akira; Yan, Jie

    2013-01-01

    Dan is a transcription factor that regulates the ttd operon encoding tartrate dehydratase. During anaerobic conditions, its copy number increases by 100-fold, making Dan an abundant nucleoid-associated protein. However, little is known about the mode of Dan–DNA interaction. To understand its cellular functions, we used single-molecule manipulation and imaging techniques to show that Dan binds cooperatively along DNA, resulting in formation of a rigid periodic nucleoprotein filament that strongly restricts accessibility to DNA. Furthermore, in the presence of physiologic levels of magnesium, these filaments interact with each other to cause global DNA condensation. Overall, these results shed light on the architectural role of Dan in the compaction of Escherichia coli chromosomal DNA under anaerobic conditions. Formation of the nucleoprotein filament provides a basis in understanding how Dan may play roles in both chromosomal DNA protection and gene regulation. PMID:23180762

  5. A Model of Filamentous Cyanobacteria Leading to Reticulate Pattern Formation

    PubMed Central

    Tamulonis, Carlos; Kaandorp, Jaap

    2014-01-01

    The filamentous cyanobacterium, Pseudanabaena, has been shown to produce reticulate patterns that are thought to be the result of its gliding motility. Similar fossilized structures found in the geological record constitute some of the earliest signs of life on Earth. It is difficult to tie these fossils, which are billions of years old, directly to the specific microorganisms that built them. Identifying the physicochemical conditions and microorganism properties that lead microbial mats to form macroscopic structures can lead to a better understanding of the conditions on Earth at the dawn of life. In this article, a cell-based model is used to simulate the formation of reticulate patterns in cultures of Pseudanabaena. A minimal system of long and flexible trichomes capable of gliding motility is shown to be sufficient to produce stable patterns consisting of a network of streams. Varying model parameters indicate that systems with little to no cohesion, high trichome density and persistent movement are conducive to reticulate pattern formation, in conformance with experimental observations. PMID:25370380

  6. The More the Tubular: Dynamic Bundling of Actin Filaments for Membrane Tube Formation.

    PubMed

    Weichsel, Julian; Geissler, Phillip L

    2016-07-01

    Tubular protrusions are a common feature of living cells, arising from polymerization of stiff protein filaments against a comparably soft membrane. Although this process involves many accessory proteins in cells, in vitro experiments indicate that similar tube-like structures can emerge without them, through spontaneous bundling of filaments mediated by the membrane. Using theory and simulation of physical models, we have elaborated how nonequilibrium fluctuations in growth kinetics and membrane shape can yield such protrusions. Enabled by a new grand canonical Monte Carlo method for membrane simulation, our work reveals a cascade of dynamical transitions from individually polymerizing filaments to highly cooperatively growing bundles as a dynamical bottleneck to tube formation. Filament network organization as well as adhesion points to the membrane, which bias filament bending and constrain membrane height fluctuations, screen the effective attractive interactions between filaments, significantly delaying bundling and tube formation.

  7. Impact of tropomyosin isoform composition on fast skeletal muscle thin filament regulation and force development.

    PubMed

    Scellini, B; Piroddi, N; Flint, G V; Regnier, M; Poggesi, C; Tesi, C

    2015-02-01

    Tropomyosin (Tm) plays a central role in the regulation of muscle contraction and is present in three main isoforms in skeletal and cardiac muscles. In the present work we studied the functional role of α- and βTm on force development by modifying the isoform composition of rabbit psoas skeletal muscle myofibrils and of regulated thin filaments for in vitro motility measurements. Skeletal myofibril regulatory proteins were extracted (78%) and replaced (98%) with Tm isoforms as homogenous ααTm or ββTm dimers and the functional effects were measured. Maximal Ca(2+) activated force was the same in ααTm versus ββTm myofibrils, but ββTm myofibrils showed a marked slowing of relaxation and an impairment of regulation under resting conditions compared to ααTm and controls. ββTm myofibrils also showed a significantly shorter slack sarcomere length and a marked increase in resting tension. Both these mechanical features were almost completely abolished by 10 mM 2,3-butanedione 2-monoxime, suggesting the presence of a significant degree of Ca(2+)-independent cross-bridge formation in ββTm myofibrils. Finally, in motility assay experiments in the absence of Ca(2+) (pCa 9.0), complete regulation of thin filaments required greater ββTm versus ααTm concentrations, while at full activation (pCa 5.0) no effect was observed on maximal thin filament motility speed. We infer from these observations that high contents of ββTm in skeletal muscle result in partial Ca(2+)-independent activation of thin filaments at rest, and longer-lasting and less complete tension relaxation following Ca(2+) removal.

  8. The importance of corona generation and leader formation during laser filament guided discharges in air

    NASA Astrophysics Data System (ADS)

    Schmitt-Sody, Andreas; French, David; White, William; Lucero, Adrian; Roach, William P.; Hasson, Victor

    2015-03-01

    Images taken with an intensified CCD camera show the dynamics during filament guided discharge events. The images reveal that filament initiated corona plays a role in the presented results. Furthermore, the images show the formation of leaders, propagating and eventually bridging the gap between the high voltage (HV) electrodes. Analysis of the images and comparison to oscilloscope traces of voltage and current dynamics reveal the origin of the delay between the filament and HV discharge and allows for a probability of discharge analysis.

  9. STARBURST-DRIVEN GALACTIC WINDS: FILAMENT FORMATION AND EMISSION PROCESSES

    SciTech Connect

    Cooper, Jackie L.; Bicknell, Geoffrey V.; Sutherland, Ralph S.; Bland-Hawthorn, Joss

    2009-09-20

    We have performed a series of three-dimensional simulations of the interaction of a supersonic wind with a nonspherical radiative cloud. These simulations are motivated by our recent three-dimensional model of a starburst-driven galactic wind interacting with an inhomogeneous disk, which shows that an optically emitting filament can be formed by the breakup and acceleration of a cloud into a supersonic wind. In this study, we consider the evolution of a cloud with two different geometries (fractal and spherical) and investigate the importance of radiative cooling on the cloud's survival. We have also undertaken a comprehensive resolution study in order to ascertain the effect of the assumed numerical resolution on the results. We find that the ability of the cloud to radiate heat is crucial for its survival, with a radiative cloud experiencing a lower degree of acceleration and having a higher relative Mach number to the flow than in the adiabatic case. This diminishes the destructive effect of the Kelvin-Helmholtz instability on the cloud. While an adiabatic cloud is destroyed over a short period of time, a radiative cloud is broken up via the Kelvin-Helmholtz instability into numerous small, dense cloudlets, which are drawn into the flow to form a filamentary structure. The degree of fragmentation is highly dependent on the resolution of the simulation, with the number of cloudlets formed increasing as the Kelvin-Helmholtz instability is better resolved. Nevertheless, there is a clear qualitative trend, with the filamentary structure still persistent at high resolution. The geometry of the cloud affects the speed at which the cloud fragments; a wind more rapidly breaks up the cloud in regions of least density. A cloud with a more inhomogeneous density distribution fragments faster than a cloud with a more uniform structure (e.g., a sphere). We confirm the mechanism behind the formation of the Halpha emitting filaments found in our global simulations of a

  10. Allosteric regulation by cooperative conformational changes of actin filaments drives mutually exclusive binding with cofilin and myosin.

    PubMed

    Ngo, Kien Xuan; Umeki, Nobuhisa; Kijima, Saku T; Kodera, Noriyuki; Ueno, Hiroaki; Furutani-Umezu, Nozomi; Nakajima, Jun; Noguchi, Taro Q P; Nagasaki, Akira; Tokuraku, Kiyotaka; Uyeda, Taro Q P

    2016-10-20

    Heavy meromyosin (HMM) of myosin II and cofilin each binds to actin filaments cooperatively and forms clusters along the filaments, but it is unknown whether the two cooperative bindings are correlated and what physiological roles they have. Fluorescence microscopy demonstrated that HMM-GFP and cofilin-mCherry each bound cooperatively to different parts of actin filaments when they were added simultaneously in 0.2 μM ATP, indicating that the two cooperative bindings are mutually exclusive. In 0.1 mM ATP, the motor domain of myosin (S1) strongly inhibited the formation of cofilin clusters along actin filaments. Under this condition, most actin protomers were unoccupied by S1 at any given moment, suggesting that transiently bound S1 alters the structure of actin filaments cooperatively and/or persistently to inhibit cofilin binding. Consistently, cosedimentation experiments using copolymers of actin and actin-S1 fusion protein demonstrated that the fusion protein affects the neighboring actin protomers, reducing their affinity for cofilin. In reciprocal experiments, cofilin-actin fusion protein reduced the affinity of neighboring actin protomers for S1. Thus, allosteric regulation by cooperative conformational changes of actin filaments contributes to mutually exclusive cooperative binding of myosin II and cofilin to actin filaments, and presumably to the differential localization of both proteins in cells.

  11. Allosteric regulation by cooperative conformational changes of actin filaments drives mutually exclusive binding with cofilin and myosin

    PubMed Central

    Ngo, Kien Xuan; Umeki, Nobuhisa; Kijima, Saku T.; Kodera, Noriyuki; Ueno, Hiroaki; Furutani-Umezu, Nozomi; Nakajima, Jun; Noguchi, Taro Q. P.; Nagasaki, Akira; Tokuraku, Kiyotaka; Uyeda, Taro Q. P.

    2016-01-01

    Heavy meromyosin (HMM) of myosin II and cofilin each binds to actin filaments cooperatively and forms clusters along the filaments, but it is unknown whether the two cooperative bindings are correlated and what physiological roles they have. Fluorescence microscopy demonstrated that HMM-GFP and cofilin-mCherry each bound cooperatively to different parts of actin filaments when they were added simultaneously in 0.2 μM ATP, indicating that the two cooperative bindings are mutually exclusive. In 0.1 mM ATP, the motor domain of myosin (S1) strongly inhibited the formation of cofilin clusters along actin filaments. Under this condition, most actin protomers were unoccupied by S1 at any given moment, suggesting that transiently bound S1 alters the structure of actin filaments cooperatively and/or persistently to inhibit cofilin binding. Consistently, cosedimentation experiments using copolymers of actin and actin-S1 fusion protein demonstrated that the fusion protein affects the neighboring actin protomers, reducing their affinity for cofilin. In reciprocal experiments, cofilin-actin fusion protein reduced the affinity of neighboring actin protomers for S1. Thus, allosteric regulation by cooperative conformational changes of actin filaments contributes to mutually exclusive cooperative binding of myosin II and cofilin to actin filaments, and presumably to the differential localization of both proteins in cells. PMID:27762277

  12. Small RNAs regulate plant responses to filamentous pathogens.

    PubMed

    Kuan, Tung; Zhai, Yi; Ma, Wenbo

    2016-08-01

    Small RNAs are central players of RNA silencing in eukaryotes. These short RNA molecules (20-25 nucleotides in length) repress target gene expression based on sequence complementarity. While small RNAs are well-known for their essential function in regulating growth and development, recent research has revealed that they also influence plant immunity. Extensive changes in small RNA accumulation have been observed during infection. This review focuses on specific small RNA changes that are involved in plant responses to filamentous eukaryotic pathogens including fungi and oomycetes. We describe how changes in small RNA accumulation influence plant immunity and summarize the cellular processes affected by these small RNAs. In particular, we discuss secondary small interfering RNAs that directly modulate the expression of defense-related genes.

  13. A Simple Dynamical Model for Filament Formation in the Solar Corona

    NASA Astrophysics Data System (ADS)

    Litvinenko, Y.

    2005-12-01

    Filament formation in the solar corona is considered in the case of a slowly evolving force-free magnetic field. The strong-field approximation is used, which takes into account the magnetohydrodynamic equations of motion, induction, and compressibility. Methods for solving the relevant equations are presented and applied to filament modeling. A three-dimensional calculation is presented, which uses linear force-free magnetic fields. The boundary conditions are chosen to resemble the qualitative linkage model for the formation of filaments, suggested by Martens and Zwaan (2001). Consistent with this model, dense formations, reminiscent of filament pillars, are shown to appear in the corona above the region of converging and canceling magnetic bipoles. The results demonstrate the principal role of magnetic field in the dynamical processes of dense plasma accumulation and support in filaments. The model can be useful for clarifying the role of flux emergence in coronal mass ejection initiation.

  14. Filament formation in wind-cloud interactions - I. Spherical clouds in uniform magnetic fields

    NASA Astrophysics Data System (ADS)

    Banda-Barragán, W. E.; Parkin, E. R.; Federrath, C.; Crocker, R. M.; Bicknell, G. V.

    2016-01-01

    Filamentary structures are ubiquitous in the interstellar medium, yet their formation, internal structure, and longevity have not been studied in detail. We report the results from a comprehensive numerical study that investigates the characteristics, formation, and evolution of filaments arising from magnetohydrodynamic interactions between supersonic winds and dense clouds. Here, we improve on previous simulations by utilizing sharper density contrasts and higher numerical resolutions. By following multiple density tracers, we find that material in the envelopes of the clouds is removed and deposited downstream to form filamentary tails, while the cores of the clouds serve as footpoints and late-stage outer layers of these tails. Aspect ratios ≳12, subsonic velocity dispersions ˜0.1-0.3 of the wind sound speed, and magnetic field amplifications ˜100 are found to be characteristic of these filaments. We also report the effects of different magnetic field strengths and orientations. The magnetic field strength regulates vorticity production: sinuous filamentary towers arise in non-magnetic environments, while strong magnetic fields inhibit small-scale perturbations at boundary layers making tails less turbulent. Magnetic field components aligned with the direction of the flow favour the formation of pressure-confined flux ropes inside the tails, whilst transverse components tend to form current sheets. Softening the equation of state to nearly isothermal leads to suppression of dynamical instabilities and further collimation of the tail. Towards the final stages of the evolution, we find that small cloudlets and distorted filaments survive the break-up of the clouds and become entrained in the winds, reaching velocities ˜0.1 of the wind speed.

  15. An Observational Study of the Recurring Formation and Dissipation of a Dynamic Filament

    NASA Astrophysics Data System (ADS)

    Zhou, Guiping; Wang, Jingxiu; Zhang, Jie

    2016-10-01

    Based on observations at the Hα wavelength from the Hinode spacecraft, we report here the detailed process of a dynamical filament that showed repeated appearance and dissipation in a filament channel. First, Hα short fibrils spreading in the pre-formed filament channel joined into longer threads. The joining process was found to be accompanied by small-scale brightening activity, indicating the possible involvement of magnetic reconnection. The forming filament was thickened by merging the neighboring dark threads that were nearly parallel to the axis and also those adjacent to its main endpoints. The formed filament as a single coherent structure only existed for tens of minutes, immediately followed by the dissipation. The dissipation appeared to start with expansion of the filament body, ascending and stripping away of the filament threads, and mass drainage along the legs of the filament. The formation-dissipation process of the filament was repeated three times within the four-hour observational window of Hinode. These observations indicate that the filament structure is highly dynamic. This study provides the observational evidence to confirm the hypothesis of Martin et al. ( Ann. Geophys. 26, 3061, 2008) on the irreversible build-up of magnetic fields in the corona by discrete threads or groups of threads ascending bodily into the corona.

  16. CARMA Large Area Star Formation Survey: Observational Analysis of Filaments in the Serpens South Molecular Cloud

    NASA Astrophysics Data System (ADS)

    Fernández-López, M.; Arce, H. G.; Looney, L.; Mundy, L. G.; Storm, S.; Teuben, P. J.; Lee, K.; Segura-Cox, D.; Isella, A.; Tobin, J. J.; Rosolowsky, E.; Plunkett, A.; Kwon, W.; Kauffmann, J.; Ostriker, E.; Tassis, K.; Shirley, Y. L.; Pound, M.

    2014-08-01

    We present the N2H+ (J = 1 → 0) map of the Serpens South molecular cloud obtained as part of the CARMA Large Area Star Formation Survey. The observations cover 250 arcmin2 and fully sample structures from 3000 AU to 3 pc with a velocity resolution of 0.16 km s-1, and they can be used to constrain the origin and evolution of molecular cloud filaments. The spatial distribution of the N2H+ emission is characterized by long filaments that resemble those observed in the dust continuum emission by Herschel. However, the gas filaments are typically narrower such that, in some cases, two or three quasi-parallel N2H+ filaments comprise a single observed dust continuum filament. The difference between the dust and gas filament widths casts doubt on Herschel ability to resolve the Serpens South filaments. Some molecular filaments show velocity gradients along their major axis, and two are characterized by a steep velocity gradient in the direction perpendicular to the filament axis. The observed velocity gradient along one of these filaments was previously postulated as evidence for mass infall toward the central cluster, but these kind of gradients can be interpreted as projection of large-scale turbulence.

  17. Chaperonin filaments : their formation and an evaluation of methods for studying them.

    SciTech Connect

    Yaoi, T.; Kagawa, K. H.; Trent, J. D.; Center for Mechanistic Biology and Biotechnology

    1998-08-01

    Chaperonins are multisubunit protein complexes that can be isolated from cells as high-molecular-weight structures that appear as double rings in the electron microscope. We recently discovered that chaperonin double rings isolated from the hyperthermophilic archaeon Sulfolobus shibatae, when incubated at physiological temperatures in the presence of ATP and Mg{sup 2+}, stacked into filaments; we hypothesized that these filaments are related to filaments seen inside S. shibatae cells and that chaperonins exist as filaments in vivo. This paper elucidates the conditions under which we have observed S. shibatae chaperonins to form filaments and evaluates native polyacrylamide gel electrophoresis (PAGE), TEM, spectrophotometry, and centrifugation as methods for studying these filaments. We observed that in the presence of Mg{sup 2+} combined with ATP, ADP, ATP{gamma}S, or GTP, native PAGE indicated that chaperonin subunits assembled into double rings and that the conformation of these double rings was effected by nucleotide binding, but we saw no indication of chaperonin filament formation. Under these same conditions, however, TEM, spectroscopy, and centrifugation methods indicated that chaperonin subunits and double rings had assembled into filaments. We determined that this discrepancy in the representation of the chaperonin structure was due to the native PAGE method itself. When we exposed chaperonin filaments to the electrophoretic field used in native PAGE, the filaments dissociated into double rings. This suggests that TEM, spectrophotometry, and centrifugation are the preferred methods for studying the higher-order structures of chaperonins, which are likely to be of biological significance.

  18. Tetrahedral collapse: a rotational toy model of simultaneous dark-matter halo, filament and wall formation

    NASA Astrophysics Data System (ADS)

    Neyrinck, Mark C.

    2016-07-01

    We discuss an idealized model of halo formation, in which a collapsing halo node is tetrahedral, with a filament extruding from each of its four faces, and with a wall connecting each pair of filaments. In the model, filaments generally spin when they form, and the halo spins if and only if there is some rotation in filaments. This is the simplest possible fully three-dimensional halo collapse in the `origami approximation', in which voids are irrotational, and the dark-matter sheet out of which dark-matter structures form is allowed to fold in position-velocity phase space, but not stretch (i.e. it cannot vary in density along a stream). Up to an overall scaling, the four filament directions, and only three other quantities, such as filament spins, suffice to determine all of the collapse's properties: the shape, mass, and spin of the halo; the densities per unit length and spins of all filaments; and masses per unit area of the walls. If the filaments are arranged regular-tetrahedrally, filament properties obey simple laws, reminiscent of angular-momentum conservation. The model may be most useful in understanding spin correlations between neighbouring galaxies joined by filaments; these correlations would give intrinsic alignments between galaxies, essential to understand for accurate cosmological weak-lensing measurements.

  19. Bypass of Candida albicans Filamentation/Biofilm Regulators through Diminished Expression of Protein Kinase Cak1

    PubMed Central

    Xu, Wenjie; Aleynikov, Tatyana; Cullen, Paul J.; Lanni, Frederick; Andes, David R.

    2016-01-01

    Biofilm formation on implanted medical devices is a major source of lethal invasive infection by Candida albicans. Filamentous growth of this fungus is tied to biofilm formation because many filamentation-associated genes are required for surface adherence. Cell cycle or cell growth defects can induce filamentation, but we have limited information about the coupling between filamentation and filamentation-associated gene expression after cell cycle/cell growth inhibition. Here we identified the CDK activating protein kinase Cak1 as a determinant of filamentation and filamentation-associated gene expression through a screen of mutations that diminish expression of protein kinase-related genes implicated in cell cycle/cell growth control. A cak1 diminished expression (DX) strain displays filamentous growth and expresses filamentation-associated genes in the absence of typical inducing signals. In a wild-type background, expression of filamentation-associated genes depends upon the transcription factors Bcr1, Brg1, Efg1, Tec1, and Ume6. In the cak1 DX background, the dependence of filamentation-associated gene expression on each transcription factor is substantially relieved. The unexpected bypass of filamentation-associated gene expression activators has the functional consequence of enabling biofilm formation in the absence of Bcr1, Brg1, Tec1, Ume6, or in the absence of both Brg1 and Ume6. It also enables filamentous cell morphogenesis, though not biofilm formation, in the absence of Efg1. Because these transcription factors are known to have shared target genes, we suggest that cell cycle/cell growth limitation leads to activation of several transcription factors, thus relieving dependence on any one. PMID:27935965

  20. Galactic cold cores. VII. Filament formation and evolution: Methods and observational constraints

    NASA Astrophysics Data System (ADS)

    Rivera-Ingraham, A.; Ristorcelli, I.; Juvela, M.; Montillaud, J.; Men'shchikov, A.; Malinen, J.; Pelkonen, V.-M.; Marston, A.; Martin, P. G.; Pagani, L.; Paladini, R.; Paradis, D.; Ysard, N.; Ward-Thompson, D.; Bernard, J.-P.; Marshall, D. J.; Montier, L.; Tóth, L. V.

    2016-06-01

    Context. The association of filaments with protostellar objects has made these structures a priority target in star formation studies. However, little is known about the link between filament properties and their local environment. Aims: The datasets from the Herschel Galactic Cold cores key programme allow for a statistical study of filaments with a wide range of intrinsic and environmental characteristics. Characterisation of this sample can therefore be used to identify key physical parameters and quantify the role of the environment in the formation of supercritical filaments. These results are necessary to constrain theoretical models of filament formation and evolution. Methods: Filaments were extracted from fields at distance D< 500 pc with the getfilaments algorithm and characterised according to their column density profiles and intrinsic properties. Each profile was fitted with a beam-convolved Plummer-like function, and the filament structure was quantified based on the relative contributions from the filament "core", represented by a Gaussian, and "wing" component, dominated by the power-law behaviour of the Plummer-like function. These filament parameters were examined for populations associated with different background levels. Results: Filaments increase their core (Mline,core) and wing (Mline,wing) contributions while increasing their total linear mass density (Mline,tot). Both components appear to be linked to the local environment, with filaments in higher backgrounds having systematically more massive Mline,core and Mline,wing. This dependence on the environment supports an accretion-based model of filament evolution in the local neighbourhood (D ≤ 500 pc). Structures located in the highest backgrounds develop the highest central AV, Mline,core, and Mline,wing as Mline,tot increases with time, favoured by the local availability of material and the enhanced gravitational potential. Our results indicate that filaments acquiring a significantly

  1. [Epigenetic regulation of secondary metabolite biosynthesis in filamentous fungi: a review].

    PubMed

    Zhou, Rui; Liao, Guojian; Hu, Changhua

    2011-08-01

    Secondary metabolites of filamentous fungi are important sources of new drugs, and their biosynthetic processes are regulated by numerous factors. Recent studies indicate that many filamentous fungal secondary metabolites are regulated by epigenetic modifications, which not only affect the titers of secondary metabolites, but also activate the cryptic gene clusters. This review summarizes recent advances of epigenetic application in filamentous fungal secondary metabolite biosynthesis, especially the types of fungal epigenetic modification and epigenetic remodeling of the fungal secondary metabolism. The application of epigenetic theory in filamentous fungi is becoming a new strategy for fungal strain improvement and a powerful method to obtain novel natural products.

  2. Bcr1 plays a central role in the regulation of opaque cell filamentation in Candida albicans

    PubMed Central

    Guan, Guobo; Xie, Jing; Tao, Li; Nobile, Clarissa J.; Sun, Yuan; Cao, Chengjun; Tong, Yaojun; Huang, Guanghua

    2013-01-01

    Summary The human fungal pathogen Candida albicans has at least two types of morphological transitions: white to opaque cell transitions and yeast to hyphal transitions. Opaque cells have historically not been known to undergo filamentation under standard filament-inducing conditions. Here we find that Bcr1 and its downstream regulators Cup9, Nrg1 and Czf1 and the cAMP-signaling pathway control opaque cell filamentation in C. albicans. We have shown that deletion of BCR1, CUP9, NRG1 and CZF1 results in opaque cell filamentation under standard culture conditions. Disruption of BCR1 in white cells has no obvious effect on hyphal growth, suggesting that Bcr1 is an opaque-specific regulator of filamentation under the conditions tested. Moreover, inactivation of the cAMP-signaling pathway or disruption of its downstream transcriptional regulators, FLO8 and EFG1, strikingly attenuates filamentation in opaque cells of the bcr1/bcr1 mutant. Deletion of HGC1, a downstream gene of the cAMP-signaling pathway encoding G1 cyclin-related protein, completely blocks opaque cell filamentation induced by inactivation of BCR1. These results demonstrate that Bcr1 regulated opaque cell filamentation is dependent on the cAMP-signaling pathway. This study establishes a link between the white-opaque switch and the yeast-filamentous growth transition in C. albicans. PMID:23808664

  3. Bcr1 plays a central role in the regulation of opaque cell filamentation in Candida albicans.

    PubMed

    Guan, Guobo; Xie, Jing; Tao, Li; Nobile, Clarissa J; Sun, Yuan; Cao, Chengjun; Tong, Yaojun; Huang, Guanghua

    2013-08-01

    The human fungal pathogen Candida albicans has at least two types of morphological transitions: white to opaque cell transitions and yeast to hyphal transitions. Opaque cells have historically not been known to undergo filamentation under standard filament-inducing conditions. Here we find that Bcr1 and its downstream regulators Cup9, Nrg1 and Czf1 and the cAMP-signalling pathway control opaque cell filamentation in C. albicans. We have shown that deletion of BCR1, CUP9, NRG1 and CZF1 results in opaque cell filamentation under standard culture conditions. Disruption of BCR1 in white cells has no obvious effect on hyphal growth, suggesting that Bcr1 is an opaque-specific regulator of filamentation under the conditions tested. Moreover, inactivation of the cAMP-signalling pathway or disruption of its downstream transcriptional regulators, FLO8 and EFG1, strikingly attenuates filamentation in opaque cells of the bcr1/bcr1 mutant. Deletion of HGC1, a downstream gene of the cAMP-signalling pathway encoding G1 cyclin-related protein, completely blocks opaque cell filamentation induced by inactivation of BCR1. These results demonstrate that Bcr1 regulated opaque cell filamentation is dependent on the cAMP-signalling pathway. This study establishes a link between the white-opaque switch and the yeast-filamentous growth transition in C. albicans. © 2013 John Wiley & Sons Ltd.

  4. Laser-filamentation-induced condensation and snow formation in a cloud chamber.

    PubMed

    Ju, Jingjing; Liu, Jiansheng; Wang, Cheng; Sun, Haiyi; Wang, Wentao; Ge, Xiaochun; Li, Chuang; Chin, See Leang; Li, Ruxin; Xu, Zhizhan

    2012-04-01

    Using 1 kHz, 9 mJ femtosecond laser pulses, we demonstrate laser-filamentation-induced spectacular snow formation in a cloud chamber. An intense updraft of warm moist air is generated owing to the continuous heating by the high-repetition filamentation. As it encounters the cold air above, water condensation and large-sized particles spread unevenly across the whole cloud chamber via convection and cyclone like action on a macroscopic scale. This indicates that high-repetition filamentation plays a significant role in macroscopic laser-induced water condensation and snow formation.

  5. THE FORMATION AND MAGNETIC STRUCTURES OF ACTIVE-REGION FILAMENTS OBSERVED BY NVST, SDO, AND HINODE

    SciTech Connect

    Yan, X. L.; Xue, Z. K.; Wang, J. C.; Xiang, Y. Y.; Kong, D. F.; Yang, L. H.; Pan, G. M.

    2015-08-15

    To better understand the properties of solar active-region filaments, we present a detailed study on the formation and magnetic structures of two active-region filaments in active region NOAA 11884 during a period of four days. It is found that the shearing motion of the opposite magnetic polarities and the rotation of the small sunspots with negative polarity play an important role in the formation of two active-region filaments. During the formation of these two active-region filaments, one foot of the filaments was rooted in a small sunspot with negative polarity. The small sunspot rotated not only around another small sunspot with negative polarity, but also around the center of its umbra. By analyzing the nonlinear force-free field extrapolation using the vector magnetic fields in the photosphere, twisted structures were found in the two active-region filaments prior to their eruptions. These results imply that the magnetic fields were dragged by the shearing motion between opposite magnetic polarities and became more horizontal. The sunspot rotation twisted the horizontal magnetic fields and finally formed the twisted active-region filaments.

  6. Regulation of actin filament length in erythrocytes and striated muscle.

    PubMed

    Fowler, V M

    1996-02-01

    Actin filaments polymerize in vitro to lengths which display an exponential distribution, yet in many highly differentiated cells they can be precisely maintained at uniform lengths in elaborate supramolecular structures. Recent results obtained using two classic model systems, the erythrocyte membrane cytoskeleton and the striated muscle sarcomere, reveal surprising similarities and instructive differences in the molecules and mechanisms responsible for determining and maintaining actin filament lengths in these two systems. Tropomodulin caps the slow-growing, pointed filament ends in muscle and in erythrocytes. CapZ caps the fast-growing, barbed filament ends in striated muscle, whereas a newly discovered barbed end capping protein, adducin, may cap the barbed filament ends in erythrocytes. The mechanisms responsible for specifying the characteristic filament lengths in these systems are more elusive and may include strict control of the relative amounts of actin filament capping proteins and side-binding proteins, molecular templates (e.g. tropomyosin and nebulin) and/or verniers (e.g. tropomyosin).

  7. Arabidopsis FIM5 decorates apical actin filaments and regulates their organization in the pollen tube

    PubMed Central

    Zhang, Meng; Zhang, Ruihui; Qu, Xiaolu; Huang, Shanjin

    2016-01-01

    The actin cytoskeleton is increasingly recognized as a major regulator of pollen tube growth. Actin filaments have distinct distribution patterns and dynamic properties within different regions of the pollen tube. Apical actin filaments are highly dynamic and crucial for pollen tube growth. However, how apical actin filaments are generated and properly constructed remains an open question. Here we showed that Arabidopsis fimbrin5 (FIM5) decorates filamentous structures throughout the entire tube but is apically concentrated. Apical actin structures are disorganized to different degrees in the pollen tubes of fim5 loss-of-function mutants. Further observations suggest that apical actin structures are not constructed properly because apical actin filaments cannot be maintained at the cortex of fim5 pollen tubes. Actin filaments appeared to be more curved in fim5 pollen tubes and this was confirmed by measurements showing that the convolutedness and the rate of change of convolutedness of actin filaments was significantly increased in fim5 pollen tubes. This suggests that the rigidity of the actin filaments may be compromised in fim5 pollen tubes. Further, the apical cell wall composition is altered, implying that tip-directed vesicle trafficking events are impaired in fim5 pollen tubes. Thus, we found that FIM5 decorates apical actin filaments and regulates their organization in order to drive polarized pollen tube growth. PMID:27117336

  8. A Simple Dynamical Model for Filament Formation in the Solar Corona

    NASA Astrophysics Data System (ADS)

    Litvinenko, Yuri E.; Wheatland, M. S.

    2005-09-01

    Filament formation in the solar atmosphere is considered. In the limit of sub-Alfvénic but supersonic motion, plasma flow in the solar corona is driven via the induction equation by a slow evolution of force-free magnetic fields. Methods for solving the relevant magnetohydrodynamic equations are presented and applied to filament modeling in two and three dimensions. An illustrative two-dimensional example is given, which is based on a potential magnetic field with a dip. The example describes the formation of a normal filament between two bipolar regions on the Sun. Next a detailed three-dimensional calculation is presented, which uses linear force-free magnetic fields. The boundary conditions are chosen to resemble the qualitative ``head-to-tail'' linkage model for the formation of filaments, suggested by Martens & Zwaan. Consistent with this model, dense formations, reminiscent of filament pillars, are shown to appear in the corona above the region of converging and canceling magnetic bipoles. The numerical results are consistent with the principal role of magnetic field in the dynamical processes of dense plasma accumulation and support in filaments, advocated by Martens & Zwaan.

  9. Convergent Regulation of Candida albicans Aft2 and Czf1 in Invasive and Opaque Filamentation.

    PubMed

    Xu, Ning; Dong, Yi-Jie; Yu, Qi-Lin; Zhang, Bing; Zhang, Meng; Jia, Chang; Chen, Yu-Lu; Zhang, Biao; Xing, Lai-Jun; Li, Ming-Chun

    2015-09-01

    Candida albicans is the most common fungal pathogen of mucosal infections and invasive diseases in immuno-compromised humans. The abilities of yeast-hyphal growth and white-opaque switching affect C. albicans physiology and virulence. Here, we showed that C. albicans Aft2 regulator was required for embedded filamentous growth and opaque cell-type formation. Under low-temperature matrix embedded conditions, Aft2 functioned downstream of Czf1-mediated pathway and was required for invasive filamentation. Moreover, deletion of AFT2 significantly reduced opaque cell-type formation under N-acetylglucosamine (GlcNAc) inducing conditions. Ectopic expression of CZF1 slightly increased the white-opaque switching frequency in the aft2Δ/Δ mutant, but did not completely restore to wild-type levels, suggesting that Czf1 at least partially bypassed the essential requirement for Aft2 in response to opaque-inducing cues. In addition, multiple environmental cues altered AFT2 mRNA and protein levels, such as low temperature, physical environment and GlcNAc. Although the absence of Czf1 or Efg1 also increased the expression level of AFT2 gene, deletion of CZF1 remarkably reduced the stability of Aft2 protein. Furthermore, C. albicans Aft2 physically interacted with Czf1 under all tested conditions, whereas the interaction between Aft2 and Efg1 was barely detectable under embedded conditions, supporting the hypothesis that Aft2, together with Czf1, contributed to activate filamentous growth by antagonizing Efg1-mediated repression under matrix-embedded conditions.

  10. Chaperonin filaments: their formation and an evaluation of methods for studying them.

    PubMed

    Yaoi, T; Kagawa, H K; Trent, J D

    1998-08-01

    Chaperonins are multisubunit protein complexes that can be isolated from cells as high-molecular-weight structures that appear as double rings in the electron microscope. We recently discovered that chaperonin double rings isolated from the hyperthermophilic archaeon Sulfolobus shibatae, when incubated at physiological temperatures in the presence of ATP and Mg2+, stacked into filaments; we hypothesized that these filaments are related to filaments seen inside S. shibatae cells and that chaperonins exist as filaments in vivo (J. D. Trent et al., 1997, Proc. Natl. Acad. Sci. USA 94, 5383-5388). This paper elucidates the conditions under which we have observed S. shibatae chaperonins to form filaments and evaluates native polyacrylamide gel electrophoresis (PAGE), TEM, spectrophotometry, and centrifugation as methods for studying these filaments. We observed that in the presence of Mg2+ combined with ATP, ADP, ATPgammaS, or GTP, native PAGE indicated that chaperonin subunits assembled into double rings and that the conformation of these double rings was effected by nucleotide binding, but we saw no indication of chaperonin filament formation. Under these same conditions, however, TEM, spectroscopy, and centrifugation methods indicated that chaperonin subunits and double rings had assembled into filaments. We determined that this discrepancy in the representation of the chaperonin structure was due to the native PAGE method itself. When we exposed chaperonin filaments to the electrophoretic field used in native PAGE, the filaments dissociated into double rings. This suggests that TEM, spectrophotometry, and centrifugation are the preferred methods for studying the higher-order structures of chaperonins, which are likely to be of biological significance. Copyright 1998 Academic Press.

  11. Addressable Direct-Write Nanoscale Filament Formation and Dissolution by Nanoparticle-Mediated Bipolar Electrochemistry.

    PubMed

    Crouch, Garrison M; Han, Donghoon; Fullerton-Shirey, Susan K; Go, David B; Bohn, Paul W

    2017-05-23

    Nanoscale conductive filaments, usually associated with resistive memory or memristor technology, may also be used for chemical sensing and nanophotonic applications; however, realistic implementation of the technology requires precise knowledge of the conditions that control the formation and dissolution of filaments. Here we describe and characterize an addressable direct-write nanoelectrochemical approach to achieve repeatable formation/dissolution of Ag filaments across a ∼100 nm poly(ethylene oxide) (PEO) film containing either Ag(+) alone or Ag(+) together with 50 nm Ag-nanoparticles acting as bipolar electrodes. Using a conductive AFM tip, formation occurs when the PEO film is subjected to a forward bias, and dissolution occurs under reverse bias. Formation-dissolution kinetics were studied for three film compositions: Ag|PEO-Ag(+), Ag|poly(ethylene glycol) monolayer-PEO-Ag(+), and Ag|poly(ethylene glycol) monolayer-PEO-Ag(+)/Ag-nanoparticle. Statistical analysis shows that the distribution of formation times exhibits Gaussian behavior, and the fastest average initial formation time occurs for the Ag|PEO-Ag(+) system. In contrast, formation in the presence of Ag nanoparticles likely proceeds by a noncontact bipolar electrochemical mechanism, exhibiting the slowest initial filament formation. Dissolution times are log-normal for all three systems, and repeated reformation of filaments from previously formed structures is characterized by rapid regrowth. The direct-write bipolar electrochemical deposition/dissolution strategy developed here presents an approach to reconfigurable, noncontact in situ wiring of nanoparticle arrays-thereby enabling applications where actively controlled connectivity of nanoparticle arrays is used to manipulate nanoelectronic and nanophotonic behavior. The system further allows for facile manipulation of experimental conditions while simultaneously characterizing surface conditions and filament formation/dissolution kinetics.

  12. Filament formation by metabolic enzymes is a specific adaptation to an advanced state of cellular starvation

    PubMed Central

    Petrovska, Ivana; Nüske, Elisabeth; Munder, Matthias C; Kulasegaran, Gayathrie; Malinovska, Liliana; Kroschwald, Sonja; Richter, Doris; Fahmy, Karim; Gibson, Kimberley; Verbavatz, Jean-Marc; Alberti, Simon

    2014-01-01

    One of the key questions in biology is how the metabolism of a cell responds to changes in the environment. In budding yeast, starvation causes a drop in intracellular pH, but the functional role of this pH change is not well understood. Here, we show that the enzyme glutamine synthetase (Gln1) forms filaments at low pH and that filament formation leads to enzymatic inactivation. Filament formation by Gln1 is a highly cooperative process, strongly dependent on macromolecular crowding, and involves back-to-back stacking of cylindrical homo-decamers into filaments that associate laterally to form higher order fibrils. Other metabolic enzymes also assemble into filaments at low pH. Hence, we propose that filament formation is a general mechanism to inactivate and store key metabolic enzymes during a state of advanced cellular starvation. These findings have broad implications for understanding the interplay between nutritional stress, the metabolism and the physical organization of a cell. DOI: http://dx.doi.org/10.7554/eLife.02409.001 PMID:24771766

  13. Stochastic dynamics of actin filaments in guard cells regulating chloroplast localization during stomatal movement.

    PubMed

    Wang, Xiu-Ling; Gao, Xin-Qi; Wang, Xue-Chen

    2011-08-01

    Actin filaments and chloroplasts in guard cells play roles in stomatal function. However, detailed actin dynamics vary, and the roles that they play in chloroplast localization during stomatal movement remain to be determined. We examined the dynamics of actin filaments and chloroplast localization in transgenic tobacco expressing green fluorescent protein (GFP)-mouse talin in guard cells by time-lapse imaging. Actin filaments showed sliding, bundling and branching dynamics in moving guard cells. During stomatal movement, long filaments can be severed into small fragments, which can form longer filaments by end-joining activities. With chloroplast movement, actin filaments near chloroplasts showed severing and elongation activity in guard cells during stomatal movement. Cytochalasin B treatment abolished elongation, bundling and branching activities of actin filaments in guard cells, and these changes of actin filaments, and as a result, more chloroplasts were localized at the centre of guard cells. However, chloroplast turning to avoid high light, and sliding of actin fragments near the chloroplast, was unaffected following cytochalasin B treatment in guard cells. We suggest that the sliding dynamics of actin may play roles in chloroplast turning in guard cells. Our results indicate that the stochastic dynamics of actin filaments in guard cells regulate chloroplast localization during stomatal movement.

  14. THE EVOLUTION OF THE ELECTRIC CURRENT DURING THE FORMATION AND ERUPTION OF ACTIVE-REGION FILAMENTS

    SciTech Connect

    Wang, Jincheng; Yan, Xiaoli; Qu, Zhongquan; Xue, Zhike; Xiang, Yongyuan; Li, Hao

    2016-02-01

    We present a comprehensive study of the electric current related to the formation and eruption of active region filaments in NOAA AR 11884. The vertical current on the solar surface was investigated by using vector magnetograms (VMs) observed by HMI on board the Solar Dynamics Observatory. To obtain the electric current along the filament's axis, we reconstructed the magnetic fields above the photosphere by using nonlinear force-free field extrapolation based on photospheric VMs. Spatio-temporal evolutions of the vertical current on the photospheric surface and the horizontal current along the filament's axis were studied during the long-term evolution and eruption-related period, respectively. The results show that the vertical currents of the entire active region behaved with a decreasing trend and the magnetic fields also kept decreasing during the long-term evolution. For the eruption-related evolution, the mean transverse field strengths decreased before two eruptions and increased sharply after two eruptions in the vicinity of the polarity inversion lines underneath the filament. The related vertical current showed different behaviors in two of the eruptions. On the other hand, a very interesting feature was found: opposite horizontal currents with respect to the current of the filament's axis appeared and increased under the filament before the eruptions and disappeared after the eruptions. We suggest that these opposite currents were carried by the new flux emerging from the photosphere bottom and might be the trigger mechanism for these filament eruptions.

  15. DAAM Is Required for Thin Filament Formation and Sarcomerogenesis during Muscle Development in Drosophila

    PubMed Central

    Molnár, Imre; Migh, Ede; Szikora, Szilárd; Kalmár, Tibor; Végh, Attila G.; Deák, Ferenc; Barkó, Szilvia; Bugyi, Beáta; Orfanos, Zacharias; Kovács, János; Juhász, Gábor; Váró, György; Nyitrai, Miklós; Sparrow, John; Mihály, József

    2014-01-01

    During muscle development, myosin and actin containing filaments assemble into the highly organized sarcomeric structure critical for muscle function. Although sarcomerogenesis clearly involves the de novo formation of actin filaments, this process remained poorly understood. Here we show that mouse and Drosophila members of the DAAM formin family are sarcomere-associated actin assembly factors enriched at the Z-disc and M-band. Analysis of dDAAM mutants revealed a pivotal role in myofibrillogenesis of larval somatic muscles, indirect flight muscles and the heart. We found that loss of dDAAM function results in multiple defects in sarcomere development including thin and thick filament disorganization, Z-disc and M-band formation, and a near complete absence of the myofibrillar lattice. Collectively, our data suggest that dDAAM is required for the initial assembly of thin filaments, and subsequently it promotes filament elongation by assembling short actin polymers that anneal to the pointed end of the growing filaments, and by antagonizing the capping protein Tropomodulin. PMID:24586196

  16. Core and filament formation in magnetized, self-gravitating isothermal layers

    SciTech Connect

    Van Loo, Sven; Keto, Eric; Zhang, Qizhou

    2014-07-01

    We examine the role of the gravitational instability in an isothermal, self-gravitating layer threaded by magnetic fields on the formation of filaments and dense cores. Using a numerical simulation, we follow the non-linear evolution of a perturbed equilibrium layer. The linear evolution of such a layer is described in the analytic work of Nagai et al. We find that filaments and dense cores form simultaneously. Depending on the initial magnetic field, the resulting filaments form either a spiderweb-like network (for weak magnetic fields) or a network of parallel filaments aligned perpendicular to the magnetic field lines (for strong magnetic fields). Although the filaments are radially collapsing, the density profile of their central region (up to the thermal scale height) can be approximated by a hydrodynamical equilibrium density structure. Thus, the magnetic field does not play a significant role in setting the density distribution of the filaments. The density distribution outside of the central region deviates from the equilibrium. The radial column density distribution is then flatter than the expected power law of r {sup –4} and similar to filament profiles observed with Herschel. Our results do not explain the near constant filament width of ∼0.1pc. However, our model does not include turbulent motions. It is expected that the accretion-driven amplification of these turbulent motions provides additional support within the filaments against gravitational collapse. Finally, we interpret the filamentary network of the massive star forming complex G14.225-0.506 in terms of the gravitational instability model and find that the properties of the complex are consistent with being formed out of an unstable layer threaded by a strong, parallel magnetic field.

  17. Characteristics and trihalomethane formation reactivity of dissolved organic matter in effluents from membrane bioreactors with and without filamentous bulking.

    PubMed

    Xia, Chufan; Ma, Defang; Gao, Baoyu; Hu, Xinxiao; Yue, Qinyan; Meng, Yingjie; Kang, Shuyu; Zhang, Bei; Qi, Yuanfeng

    2016-07-01

    In this study, synthetic wastewater was treated by two identical membrane bioreactors (MBRs): the normal sludge MBR (NS-MBR) and the bulking sludge MBR (BS-MBR). Effects of filamentous bulking on the characteristics and trihalomethane (THM) formation reactivity of MBR effluent dissolved organic matter (EfOM) were investigated. Filamentous sludge bulking had no significant influence on the regulated MBR effluent water quality except NO2-N and NO3-N. NS-MBR effluent had more low molecular weight (LMW) (<5kDa) EfOM (92.43%) than BS-MBR (75.18%). About two-thirds of EfOM from BS-MBR were hydrophilic substances. On the contrary, EfOM from NS-MBR exhibited higher hydrophobicity. The ratio of polysaccharides and proteins in MBR effluents increased after filamentous bulking. There were more protein-like materials, fulvic acid-like and humic acid-like in BS-MBR EfOM. The THM formation reactivity of BS-MBR EfOM was 30.15% of NS-MBR EfOM, whereas BS-MBR EfOM exhibited higher formation reactivity of bromine containing species.

  18. Kiloparsec-Scale Simulations of Star Formation in Disk Galaxies III. Structure and Dynamics of Filaments and Clumps in Giant Molecular Clouds

    NASA Astrophysics Data System (ADS)

    Butler, Michael J.; Tan, Jonathan C.; Van Loo, Sven

    2015-05-01

    We present hydrodynamic simulations of self-gravitating dense gas in a galactic disk, exploring scales ranging from 1 kpc down to ˜0.1 pc. Our primary goal is to understand how dense filaments form in giant molecular clouds (GMCs). These structures, often observed as infrared dark clouds (IRDCs) in the Galactic plane, are thought to be the precursors to massive stars and star clusters, so their formation may be the rate-limiting step controlling global star formation rates in galactic systems as described by the Kennicutt-Schmidt relation. Our study follows on from Van Loo et al., which carried out simulations to 0.5 pc resolution and examined global aspects of the formation of dense gas clumps and the resulting star formation rate. Here, using our higher resolution, we examine the detailed structural, kinematic, and dynamical properties of dense filaments and clumps, including mass surface density (Σ) probability distribution functions, filament mass per unit length and its dispersion, lateral Σ profiles, filament fragmentation, filament velocity gradients and infall, and degree of filament and clump virialization. Where possible, these properties are compared to observations of IRDCs. By many metrics, especially too large mass fractions of high {Σ }\\gt 1 g c{{m}-2} material, too high mass per unit length dispersion due to dense clump formation, too high velocity gradients, and too high velocity dispersion for a given mass per unit length, the simulated filaments differ from observed IRDCs. We thus conclude that IRDCs do not form from global fast collapse of GMCs. Rather, we expect that IRDC formation and collapse are slowed significantly by the influence of dynamically important magnetic fields, which may thus play a crucial role in regulating galactic star formation rates.

  19. Direct observation of conductive filament formation in Alq3 based organic resistive memories

    NASA Astrophysics Data System (ADS)

    Busby, Y.; Nau, S.; Sax, S.; List-Kratochvil, E. J. W.; Novak, J.; Banerjee, R.; Schreiber, F.; Pireaux, J.-J.

    2015-08-01

    This work explores resistive switching mechanisms in non-volatile organic memory devices based on tris(8-hydroxyquinolie)aluminum (Alq3). Advanced characterization tools are applied to investigate metal diffusion in ITO/Alq3/Ag memory device stacks leading to conductive filament formation. The morphology of Alq3/Ag layers as a function of the metal evaporation conditions is studied by X-ray reflectivity, while depth profile analysis with X-ray photoelectron spectroscopy and time-of-flight secondary ion mass spectrometry is applied to characterize operational memory elements displaying reliable bistable current-voltage characteristics. 3D images of the distribution of silver inside the organic layer clearly point towards the existence of conductive filaments and allow for the identification of the initial filament formation and inactivation mechanisms during switching of the device. Initial filament formation is suggested to be driven by field assisted diffusion of silver from abundant structures formed during the top electrode evaporation, whereas thermochemical effects lead to local filament inactivation.

  20. Direct observation of conductive filament formation in Alq3 based organic resistive memories

    SciTech Connect

    Busby, Y. Pireaux, J.-J.; Nau, S.; Sax, S.; List-Kratochvil, E. J. W.; Novak, J.; Banerjee, R.; Schreiber, F.

    2015-08-21

    This work explores resistive switching mechanisms in non-volatile organic memory devices based on tris(8-hydroxyquinolie)aluminum (Alq{sub 3}). Advanced characterization tools are applied to investigate metal diffusion in ITO/Alq{sub 3}/Ag memory device stacks leading to conductive filament formation. The morphology of Alq{sub 3}/Ag layers as a function of the metal evaporation conditions is studied by X-ray reflectivity, while depth profile analysis with X-ray photoelectron spectroscopy and time-of-flight secondary ion mass spectrometry is applied to characterize operational memory elements displaying reliable bistable current-voltage characteristics. 3D images of the distribution of silver inside the organic layer clearly point towards the existence of conductive filaments and allow for the identification of the initial filament formation and inactivation mechanisms during switching of the device. Initial filament formation is suggested to be driven by field assisted diffusion of silver from abundant structures formed during the top electrode evaporation, whereas thermochemical effects lead to local filament inactivation.

  1. Formation of conical emission of supercontinuum during filamentation of femtosecond laser radiation in fused silica

    SciTech Connect

    Kandidov, V. P. Smetanina, E. O.; Dormidonov, A. E.; Kompanets, V. O.; Chekalin, S. V.

    2011-09-15

    The formation of conical emission of supercontinuum during filamentation of femtosecond laser pulses with central wavelengths in a wide range is studied experimentally, numerically, and analytically. The frequency-angular intensity distribution of the spectral components of conical emission is determined by the interference of supercontinuum emission in a filament of a femtosecond laser pulse. The interference of supercontinuum emission has a general character, exists at different regimes of group velocity dispersion, gives rise to the fine spectral structure after the pulse splitting into subpulses and the formation of a distributed supercontinuum source in an extended filament, and causes the decomposition of the continuous spectrum of conical emission into many high-contrast maxima after pulse refocusing in the filament. In spectroscopic studies with a tunable femtosecond radiation source based on a TOPAS parametric amplifier, we used an original scheme with a wedge fused silica sample. Numerical simulations have been performed using a system of equations of nonlinear-optical interaction of laser radiation under conditions of diffraction, wave nonstationarity, and material dispersion in fused silica. The analytic study is based on the interference model of formation of conical emission by supercontinuum sources moving in a filament.

  2. Star formation in galaxies along the Pisces-Cetus Supercluster filaments

    NASA Astrophysics Data System (ADS)

    Porter, Scott C.; Raychaudhury, Somak

    2007-03-01

    We investigate the variation of current star formation in galaxies as a function of distance along three supercluster filaments, each joining pairs of rich clusters, in the Pisces-Cetus supercluster, which is part of the two-degree Field Galaxy Redshift Survey (2dFGRS). We find that even though there is a steady decline in the rate of star formation, as well as in the fraction of star-forming galaxies, as one approaches the core of a cluster at an extremity of such a filament, there is an increased activity of star formation in a narrow distance range between 3 and 4h-170 Mpc, which is 1.5-2 times the virial radius of the clusters involved. This peak in star formation is seen to be entirely due to the dwarf galaxies (-20 < MB <= -17.5). The position of the peak does not seem to depend on the velocity dispersion of the nearest cluster, undermining the importance of the gravitational effect of the clusters involved. We find that this enhancement in star formation occurs at the same place for galaxies which belong to groups within these filaments, while group members elsewhere in the 2dFGRS do not show this effect. We conclude that the most likely mechanism for this enhanced star formation is galaxy-galaxy harassment, in the crowded infalling region of rich clusters at the extremities of filaments, which induces a burst of star formation in galaxies, before they have been stripped of their gas in the denser cores of clusters. The effects of strangulation in the cores of clusters, as well as excess star formation in the infalling regions along the filaments, are more pronounced in dwarfs since they more vulnerable to the effects of strangulation and harassment than giant galaxies.

  3. STAR FORMATION IN THE TAURUS FILAMENT L 1495: FROM DENSE CORES TO STARS

    SciTech Connect

    Schmalzl, Markus; Kainulainen, Jouni; Henning, Thomas; Launhardt, Ralf; Quanz, Sascha P.; Alves, Joao; Goodman, Alyssa A.; Pineda, Jaime E.; Roman-Zuniga, Carlos G.

    2010-12-10

    We present a study of dense structures in the L 1495 filament in the Taurus Molecular Cloud and examine its star-forming properties. In particular, we construct a dust extinction map of the filament using deep near-infrared observations, exposing its small-scale structure in unprecedented detail. The filament shows highly fragmented substructures and a high mass-per-length value of M{sub line} = 17 M{sub sun} pc{sup -1}, reflecting star-forming potential in all parts of it. However, a part of the filament, namely B 211, is remarkably devoid of young stellar objects. We argue that in this region the initial filament collapse and fragmentation is still taking place and star formation is yet to occur. In the star-forming part of the filament, we identify 39 cores with masses from 0.4 to 10 M{sub sun} and preferred separations in agreement with the local Jeans length. Most of these cores exceed the Bonnor-Ebert critical mass, and are therefore likely to collapse and form stars. The dense core mass function follows a power law with exponent {Gamma} = 1.2 {+-} 0.2, a form commonly observed in star-forming regions.

  4. Tau Isoform Composition Influences Rate and Extent of Filament Formation*

    PubMed Central

    Zhong, Qi; Congdon, Erin E.; Nagaraja, Haikady N.; Kuret, Jeff

    2012-01-01

    The risk of developing tauopathic neurodegenerative disease depends in part on the levels and composition of six naturally occurring Tau isoforms in human brain. These proteins, which form filamentous aggregates in disease, vary only by the presence or absence of three inserts encoded by alternatively spliced exons 2, 3, and 10 of the Tau gene (MAPT). To determine the contribution of alternatively spliced segments to Tau aggregation propensity, the aggregation kinetics of six unmodified, recombinant human Tau isoforms were examined in vitro using electron microscopy assay methods. Aggregation propensity was then compared at the level of elementary rate constants for nucleation and extension phases. We found that all three alternatively spliced segments modulated Tau aggregation but through differing kinetic mechanisms that could synergize or compete depending on sequence context. Overall, segments encoded by exons 2 and 10 promoted aggregation, whereas the segment encoded by exon 3 depressed it with its efficacy dependent on the presence or absence of a fourth microtubule binding repeat. In general, aggregation propensity correlated with genetic risk reported for multiple tauopathies, implicating aggregation as one candidate mechanism rationalizing the correlation between Tau expression patterns and disease. PMID:22539343

  5. EMERGENCE OF HELICAL FLUX AND THE FORMATION OF AN ACTIVE REGION FILAMENT CHANNEL

    SciTech Connect

    Lites, B. W.; Kubo, M.; Berger, T.; Frank, Z.; Shine, R.; Tarbell, T.; Title, A.; Okamoto, T. J.; Otsuji, K.

    2010-07-20

    We present comprehensive observations of the formation and evolution of a filament channel within NOAA Active Region (AR) 10978 from Hinode/Solar Optical Telescope and TRACE. We employ sequences of Hinode spectro-polarimeter maps of the AR, accompanying Hinode Narrowband Filter Instrument magnetograms in the Na I D1 line, Hinode Broadband Filter Instrument filtergrams in the Ca II H line and G-band, Hinode X-ray telescope X-ray images, and TRACE Fe IX 171 A image sequences. The development of the channel resembles qualitatively that presented by Okamoto et al. in that many indicators point to the emergence of a pre-existing sub-surface magnetic flux rope. The consolidation of the filament channel into a coherent structure takes place rapidly during the course of a few hours, and the filament form then gradually shrinks in width over the following two days. Particular to this filament channel is the observation of a segment along its length of horizontal, weak (500 G) flux that, unlike the rest of the filament channel, is not immediately flanked by strong vertical plage fields of opposite polarity on each side of the filament. Because this isolated horizontal field is observed in photospheric lines, we infer that it is unlikely that the channel formed as a result of reconnection in the corona, but the low values of inferred magnetic fill fraction along the entire length of the filament channel suggest that the bulk of the field resides somewhat above the low photosphere. Correlation tracking of granulation in the G band presents no evidence for either systematic flows toward the channel or systematic shear flows along it. The absence of these flows, along with other indications of these data from multiple sources, reinforces (but does not conclusively demonstrate) the picture of an emerging flux rope as the origin of this AR filament channel.

  6. Kinetic factors determining conducting filament formation in solid polymer electrolyte based planar devices

    NASA Astrophysics Data System (ADS)

    Krishnan, Karthik; Aono, Masakazu; Tsuruoka, Tohru

    2016-07-01

    Resistive switching characteristics and conducting filament formation dynamics in solid polymer electrolyte (SPE) based planar-type atomic switches, with opposing active Ag and inert Pt electrodes, have been investigated by optimizing the device configuration and experimental parameters such as the gap distance between the electrodes, the salt inclusion in the polymer matrix, and the compliance current applied in current-voltage measurements. The high ionic conductivities of SPE enabled us to make scanning electron microscopy observations of the filament formation processes in the sub-micrometer to micrometer ranges. It was found that switching behaviour and filament growth morphology depend strongly on several kinetic factors, such as the redox reaction rate at the electrode-polymer interfaces, ion mobility in the polymer matrix, electric field strength, and the reduction sites for precipitation. Different filament formations, resulting from unidirectional and dendritic growth behaviours, can be controlled by tuning specified parameters, which in turn improves the stability and performance of SPE-based devices.Resistive switching characteristics and conducting filament formation dynamics in solid polymer electrolyte (SPE) based planar-type atomic switches, with opposing active Ag and inert Pt electrodes, have been investigated by optimizing the device configuration and experimental parameters such as the gap distance between the electrodes, the salt inclusion in the polymer matrix, and the compliance current applied in current-voltage measurements. The high ionic conductivities of SPE enabled us to make scanning electron microscopy observations of the filament formation processes in the sub-micrometer to micrometer ranges. It was found that switching behaviour and filament growth morphology depend strongly on several kinetic factors, such as the redox reaction rate at the electrode-polymer interfaces, ion mobility in the polymer matrix, electric field strength

  7. Heat-induced formation of myosin oligomer-soluble filament complex in high-salt solution.

    PubMed

    Shimada, Masato; Takai, Eisuke; Ejima, Daisuke; Arakawa, Tsutomu; Shiraki, Kentaro

    2015-02-01

    Heat-induced aggregation of myosin into an elastic gel plays an important role in the water-holding capacity and texture of meat products. Here, we investigated thermal aggregation of porcine myosin in high-salt solution over a wide temperature range by dynamic light scattering experiments. The myosin samples were readily dissolved in 1.0 M NaCl at 25 °C followed by dilution into various salt concentrations. The diluted solutions consistently contained both myosin monomers and soluble filaments. The filament size decreased with increasing salt concentration and temperature. High temperatures above Tm led to at least partial dissociation of soluble filaments and thermal unfolding, resulting in the formation of soluble oligomers and binding to the persistently present soluble filaments. Such a complex formation between the oligomers and filaments has never been observed. Our results provide new insight into the heat-induced myosin gelation in high-salt solution. Copyright © 2014 Elsevier B.V. All rights reserved.

  8. Kink deformation of Weibel-mediated current filaments and onset of shock formation

    NASA Astrophysics Data System (ADS)

    Ruyer, Charles; Alves, E. Paulo; Fiuza, Frederico

    2016-10-01

    The Weibel instability is believed to mediate the interaction of high Mach number collisionless shocks in weakly magnetized astrophysical environments. Although the generation of current filaments and strong magnetic fields by this instability has now been demonstrated experimentally, it is still not clear what is the long-term evolution of these filaments and how they lead to shock formation. We have studied the stability of Weibel-mediated current filaments using 2D/3D Particle-In-Cell simulations and analytical theory. We show that these are prone to kink-like instabilities that we characterize in both the linear and non-linear stage for a single filament, leading to an efficient ion slowing down and isotropization. We then demonstrate that our results are relevant to the self-consistent counter-streaming plasma interaction. Our 3D simulations show that the kink deformation dominates the late-stage of the interaction, when the current filaments break and most of the flow dissipation occurs, leading to the onset of magnetic turbulence and shock formation. We will discuss the important implications of these results for the shock structure and its ability to accelerate particles. This work was supported by the DOE Office of Science, Fusion Energy Science (FWP 100182).

  9. THE IMPACT OF THERMODYNAMICS ON GRAVITATIONAL COLLAPSE: FILAMENT FORMATION AND MAGNETIC FIELD AMPLIFICATION

    SciTech Connect

    Peters, Thomas; Klessen, Ralf S.; Federrath, Christoph; Smith, Rowan J.; Schleicher, Dominik R. G.; Banerjee, Robi; Sur, Sharanya

    2012-12-01

    Stars form by the gravitational collapse of interstellar gas. The thermodynamic response of the gas can be characterized by an effective equation of state. It determines how gas heats up or cools as it gets compressed, and hence plays a key role in regulating the process of stellar birth on virtually all scales, ranging from individual star clusters up to the galaxy as a whole. We present a systematic study of the impact of thermodynamics on gravitational collapse in the context of high-redshift star formation, but argue that our findings are also relevant for present-day star formation in molecular clouds. We consider a polytropic equation of state, P = k{rho}{sup {Gamma}}, with both sub-isothermal exponents {Gamma} < 1 and super-isothermal exponents {Gamma} > 1. We find significant differences between these two cases. For {Gamma} > 1, pressure gradients slow down the contraction and lead to the formation of a virialized, turbulent core. Weak magnetic fields are strongly tangled and efficiently amplified via the small-scale turbulent dynamo on timescales corresponding to the eddy-turnover time at the viscous scale. For {Gamma} < 1, on the other hand, pressure support is not sufficient for the formation of such a core. Gravitational contraction proceeds much more rapidly and the flow develops very strong shocks, creating a network of intersecting sheets and extended filaments. The resulting magnetic field lines are very coherent and exhibit a considerable degree of order. Nevertheless, even under these conditions we still find exponential growth of the magnetic energy density in the kinematic regime.

  10. ASC filament formation serves as a signal amplification mechanism for inflammasomes

    PubMed Central

    Dick, Mathias S.; Sborgi, Lorenzo; Rühl, Sebastian; Hiller, Sebastian; Broz, Petr

    2016-01-01

    A hallmark of inflammasome activation is the ASC speck, a micrometre-sized structure formed by the inflammasome adaptor protein ASC (apoptosis-associated speck-like protein containing a CARD), which consists of a pyrin domain (PYD) and a caspase recruitment domain (CARD). Here we show that assembly of the ASC speck involves oligomerization of ASCPYD into filaments and cross-linking of these filaments by ASCCARD. ASC mutants with a non-functional CARD only assemble filaments but not specks, and moreover disrupt endogenous specks in primary macrophages. Systematic site-directed mutagenesis of ASCPYD is used to identify oligomerization-deficient ASC mutants and demonstrate that ASC speck formation is required for efficient processing of IL-1β, but dispensable for gasdermin-D cleavage and pyroptosis induction. Our results suggest that the oligomerization of ASC creates a multitude of potential caspase-1 activation sites, thus serving as a signal amplification mechanism for inflammasome-mediated cytokine production. PMID:27329339

  11. The Formation of an Inverse S-shaped Active-region Filament Driven by Sunspot Motion and Magnetic Reconnection

    NASA Astrophysics Data System (ADS)

    Yan, X. L.; Priest, E. R.; Guo, Q. L.; Xue, Z. K.; Wang, J. C.; Yang, L. H.

    2016-11-01

    We present a detailed study of the formation of an inverse S-shaped filament prior to its eruption in active region NOAA 11884 from 2013 October 31 to November 2. In the initial stage, clockwise rotation of a small positive sunspot around the main negative trailing sunspot formed a curved filament. Then the small sunspot cancelled with the negative magnetic flux to create a longer active-region filament with an inverse S-shape. At the cancellation site a brightening was observed in UV and EUV images and bright material was transferred to the filament. Later the filament erupted after cancellation of two opposite polarities below the upper part of the filament. Nonlinear force-free field extrapolation of vector photospheric fields suggests that the filament may have a twisted structure, but this cannot be confirmed from the current observations.

  12. Formation and Eruption Process of a Filament in Active Region NOAA 12241

    NASA Astrophysics Data System (ADS)

    Wang, Jincheng; Yan, Xiaoli; Qu, ZhongQuan; Xue, Zhike; Yang, Liheng

    2017-04-01

    In order to better understand active-region filaments, we present an intensive study on the formation and eruption of a filament in active region NOAA 12241 during the period from 2014 December 18 to 19. Using observations from the Helioseismic and Magnetic Imager (HMI) vector magnetograms, we investigate the helicity injection rate, Lorentz force, and vertical electric current in the entire region associated with the filament. The helicity injection rate before eruption is found to be larger than that after eruption, while the vertical electric current undergoes an increase at first and then a gradual decrease, similar to what the magnetic flux undergoes. Meanwhile, we find that the right part of the filament is formed by magnetic reconnection between two bundles of magnetic field lines while the left part originated from shearing motion. The interaction of the two parts causes the eruption of this filament. The mean horizontal magnetic fields in the vicinity of the magnetic polarity inversion line (PIL) enhance rapidly during the eruption. Another striking phenomenon, where the vertical electric currents close to the magnetic PIL suddenly expand toward two sides during the eruption, is found. We propose that this fascinating feature is associated with the release of energy during the eruption.

  13. Filament formation and evolution in buoyant coastal waters: Observation and modelling

    NASA Astrophysics Data System (ADS)

    Iermano, Ilaria; Liguori, Giovanni; Iudicone, Daniele; Buongiorno Nardelli, Bruno; Colella, Simone; Zingone, Adriana; Saggiomo, Vincenzo; Ribera d'Alcalà, Maurizio

    2012-11-01

    This paper presents a detailed analysis of the formation and subsequent evolution of filament-like structures observed in a relatively small area of the mid-Tyrrhenian Sea (Mediterranean Sea). The filament dynamics and potential impact on the cross-shelf exchange budget are investigated based on a combined use of remote sensing imagery, in situ data and numerical modelling. The complexity of these phenomena is shown by focusing on four distinct events that led to cross-shelf transport, each representative of a different dynamic process and a distinct expected impact on the coastal area. A systematic analysis of available observations for the years 1998-2006 underlines the role of the interplay of atmospheric freshwater fluxes, river loads and wind stress variations, which may create favourable conditions for the convergence of shelf waters (particularly at coastal capes) and the subsequent formation of short-lived filaments along the coast. The response of the buoyant coastal waters to periods of wind reversal and fluctuating freshwater discharge rates is examined through idealised Regional Ocean Modeling System (ROMS) simulations. The filaments observed in remote sensing imagery were well reproduced by the numerical exercise, where the filaments appear as organised submesoscale structures that possess high relative vorticity and develop at the river mouths or adjacent capes. In both scenarios, the filaments appear largely determined by (i) the presence of a buoyancy anomaly, (ii) the angle between the wind pulse direction and the coast and (iii) irregularities in the coastal profile. The ensemble of results suggests that the occurrence of such transient, intense structures may contribute considerably to the biological variability and cross-shelf exchange in coastal areas with similar traits.

  14. Dense core formation by fragmentation of velocity-coherent filaments in L1517

    NASA Astrophysics Data System (ADS)

    Hacar, A.; Tafalla, M.

    2011-09-01

    Context. Low-mass star-forming cores differ from their surrounding molecular cloud in turbulence, shape, and density structure. Aims: We aim to understand how dense cores form out of the less dense cloud material by studying the connection between these two regimes. Methods: We observed the L1517 dark cloud in C18O(1-0), N2H+(J = 1-0), and SO(JN = 32 - 21) with the FCRAO 14 m telescope, and in the 1.2 mm dust continuum with the IRAM 30 m telescope. Results: Most of the gas in the cloud lies in four filaments that have typical lengths of 0.5 pc. Five starless cores are embedded in these filaments and have chemical compositions indicative of different evolutionary stages. The filaments have radial profiles of C18O(1-0) emission with a central flattened region and a power-law tail, and can be fitted approximately as isothermal, pressure-supported cylinders. The filaments, in addition, are extremely quiescent. They have subsonic internal motions and are coherent in velocity over their whole length. The large-scale motions in the filaments can be used to predict the velocity inside the cores, indicating that core formation has not decoupled the dense gas kinematically from its parental material. In two filaments, these large-scale motions consist of oscillations in the velocity centroid, and a simple kinematic model suggests that they may be related to core-forming flows. Conclusions: Core formation in L1517 seems to have occurred in two steps. First, the subsonic, velocity-coherent filaments have condensed out of the more turbulent ambient cloud. Then, the cores fragmented quasi-statically and inherited the kinematics of the filaments. Turbulence dissipation has therefore occurred mostly on scales on the order of 0.5 pc or larger, and seems to have played a small role in the formation of the individual cores. Based on observations carried out with the FCRAO 14 m and IRAM 30 m telescopes. IRAM is supported by INSU/CNRS (France), MPG (Germany) and IGN (Spain).

  15. Thin filament proteins and thin filament-linked regulation of vertebrate muscle contraction.

    PubMed

    Leavis, P C; Gergely, J

    1984-01-01

    Recent developments in the field of myofibrillar proteins will be reviewed. Consideration will be given to the proteins that participate in the contractile process itself as well as to those involved in Ca-dependent regulation of striated (skeletal and cardiac) and smooth muscle. The relation of protein structure to function will be emphasized and the relation of various physiologically and histochemically defined fiber types to the proteins found in them will be discussed.

  16. Myosin II filament assemblies in the active lamella of fibroblasts: their morphogenesis and role in the formation of actin filament bundles

    PubMed Central

    1995-01-01

    The morphogenesis of myosin II structures in active lamella undergoing net protrusion was analyzed by correlative fluorescence and electron microscopy. In rat embryo fibroblasts (REF 52) microinjected with tetramethylrhodamine-myosin II, nascent myosin spots formed close to the active edge during periods of retraction and then elongated into wavy ribbons of uniform width. The spots and ribbons initially behaved as distinct structural entities but subsequently aligned with each other in a sarcomeric-like pattern. Electron microscopy established that the spots and ribbons consisted of bipolar minifilaments associated with each other at their head-containing ends and arranged in a single row in an "open" zig-zag conformation or as a "closed" parallel stack. Ribbons also contacted each other in a nonsarcomeric, network-like arrangement as described previously (Verkhovsky and Borisy, 1993. J. Cell Biol. 123:637-652). Myosin ribbons were particularly pronounced in REF 52 cells, but small ribbons and networks were found also in a range of other mammalian cells. At the edge of the cell, individual spots and open ribbons were associated with relatively disordered actin filaments. Further from the edge, myosin filament alignment increased in parallel with the development of actin bundles. In actin bundles, the actin cross-linking protein, alpha-actinin, was excluded from sites of myosin localization but concentrated in paired sites flanking each myosin ribbon, suggesting that myosin filament association may initiate a pathway for the formation of actin filament bundles. We propose that zig-zag assemblies of myosin II filaments induce the formation of actin bundles by pulling on an actin filament network and that co-alignment of actin and myosin filaments proceeds via folding of myosin II filament assemblies in an accordion-like fashion. PMID:7490299

  17. Three-dimensional Reconstruction of Tarantula Myosin Filaments Suggests How Phosphorylation May Regulate Myosin Activity

    PubMed Central

    Alamo, Lorenzo; Wriggers, Willy; Pinto, Antonio; Bártoli, Fulvia; Salazar, Leiría; Zhao, Fa-Qing; Craig, Roger; Padrón, Raúl

    2008-01-01

    Summary Muscle contraction involves the interaction of the myosin heads of the thick filaments with actin subunits of the thin filaments. Relaxation occurs when this interaction is blocked by molecular switches on these filaments. In many muscles, myosin-linked regulation involves phosphorylation of the myosin regulatory light chains (RLC). Electron microscopy of vertebrate smooth muscle myosin molecules (regulated by phosphorylation) has provided insight into the relaxed structure, revealing that myosin is switched off by intramolecular interactions between its two heads, the free-head and the blocked head. Three-dimensional reconstruction of frozen-hydrated specimens reveals that this asymmetric head interaction is also present in native thick filaments of tarantula striated muscle. Our goal here has been to elucidate the structural features of the tarantula filament involved in phosphorylation-based regulation. A new reconstruction reveals intra- and intermolecular myosin interactions in addition to those seen previously. To help interpret the interactions, we sequenced the tarantula RLC, and fitted to the reconstruction an atomic model of the myosin head that included the predicted RLC atomic structure and an S2 crystal structure. The fitting suggests an intramolecular interaction between the cardiomyopathy loop of the free-head and its own S2 and two intermolecular interactions—between the cardio-loop of the free head and the ELC of the blocked head, and between the Leu-305 - Gln-327 “interaction loop” (loop I) of the free-head and the N-terminal fragment of the RLC of the blocked-head. These interactions, added to those previously described, would help to switch off the thick filament. Molecular dynamics simulations suggest how phosphorylation could increase the helical content of the RLC N-terminus, weakening these interactions, thus releasing both heads and activating the thick filament. PMID:18951904

  18. Three-dimensional reconstruction of tarantula myosin filaments suggests how phosphorylation may regulate myosin activity.

    PubMed

    Alamo, Lorenzo; Wriggers, Willy; Pinto, Antonio; Bártoli, Fulvia; Salazar, Leiria; Zhao, Fa-Qing; Craig, Roger; Padrón, Raúl

    2008-12-26

    Muscle contraction involves the interaction of the myosin heads of the thick filaments with actin subunits of the thin filaments. Relaxation occurs when this interaction is blocked by molecular switches on these filaments. In many muscles, myosin-linked regulation involves phosphorylation of the myosin regulatory light chains (RLCs). Electron microscopy of vertebrate smooth muscle myosin molecules (regulated by phosphorylation) has provided insight into the relaxed structure, revealing that myosin is switched off by intramolecular interactions between its two heads, the free head and the blocked head. Three-dimensional reconstruction of frozen-hydrated specimens revealed that this asymmetric head interaction is also present in native thick filaments of tarantula striated muscle. Our goal in this study was to elucidate the structural features of the tarantula filament involved in phosphorylation-based regulation. A new reconstruction revealed intra- and intermolecular myosin interactions in addition to those seen previously. To help interpret the interactions, we sequenced the tarantula RLC and fitted an atomic model of the myosin head that included the predicted RLC atomic structure and an S2 (subfragment 2) crystal structure to the reconstruction. The fitting suggests one intramolecular interaction, between the cardiomyopathy loop of the free head and its own S2, and two intermolecular interactions, between the cardiac loop of the free head and the essential light chain of the blocked head and between the Leu305-Gln327 interaction loop of the free head and the N-terminal fragment of the RLC of the blocked head. These interactions, added to those previously described, would help switch off the thick filament. Molecular dynamics simulations suggest how phosphorylation could increase the helical content of the RLC N-terminus, weakening these interactions, thus releasing both heads and activating the thick filament.

  19. Filament and core formation in nearby molecular clouds: results from the CARMA Large Area Star Formation Survey

    NASA Astrophysics Data System (ADS)

    Storm, Shaye; Mundy, Lee G.; Fernández-López, Manuel; Lee, Katherine I.; Ostriker, Eve C.; Looney, Leslie; Chen, Che-Yu; Classy Collaboration

    2015-01-01

    Stars rarely form in isolation, so it is critical to understand how the parsec-scale molecular cloud environment shapes the formation of individual dense cores at the sub-0.1 pc scale. To address the pathway to core formation in a clustered environment, I co-developed the CARMA Large Area Star Formation Survey, which spectrally imaged dense gas tracer lines across 800 square arcminutes of the Perseus and Serpens Molecular clouds with 7'' angular resolution. There are four key results from initial papers. First, I created a new non-binary dendrogram code that shows correlation between the hierarchical complexity of dense, N2H+ (J=1-0) structures and the amount of star formation activity in a cluster. This may imply that feedback from young protostars changes the structure of dense gas within a cluster and increases the amount of high column density material. Second, we discovered strong radial velocity gradients within filaments that are an order of magnitude larger than detected axial gradients. We see similar radial gradients in filaments formed in numerical simulations of converging, turbulent flows; this suggests that the observed filaments are accreting material from an environment that is flattened at larger scales, and that they are more likely to fragment locally into cores than to support the flow of gas along the filament length. Third, we constructed two size-linewidth relations using the dendrogram-identified gas structures and our high resolution maps of the gas centroid velocity and line-of-sight velocity dispersion. The two relations show distinct behavior, and we developed a theoretical framework based on isotropic turbulence to show that they support the clustered regions being flattened (sheet-like) at parsec scales, with depths on the order 0.1-0.2 pc into the sky. Finally, we found that many filaments seen with Herschel show substructure in our high resolution maps, which implies that measuring the widths of filaments may be more complex than

  20. Formation of a rotating jet during the filament eruption on 2013 April 10-11

    NASA Astrophysics Data System (ADS)

    Filippov, B.; Srivastava, A. K.; Dwivedi, B. N.; Masson, S.; Aulanier, G.; Joshi, N. C.; Uddin, W.

    2015-07-01

    We analyse multiwavelength and multiviewpoint observations of a helically twisted plasma jet formed during a confined filament eruption on 2013 April 10-11. Given a rather large-scale event with its high spatial and temporal resolution observations, it allows us to clearly understand some new physical details about the formation and triggering mechanism of twisting jet. We identify a pre-existing flux rope associated with a sinistral filament, which was observed several days before the event. The confined eruption of the filament within a null-point topology, also known as an Eiffel tower (or inverted-Y) magnetic field configuration results in the formation of a twisted jet after the magnetic reconnection near a null point. The sign of helicity in the jet is found to be the same as that of the sign of helicity in the filament. Untwisting motion of the reconnected magnetic field lines gives rise to the accelerating plasma along the jet axis. The event clearly shows the twist injection from the pre-eruptive magnetic field to the jet.

  1. Expansion and concatenation of nonmuscle myosin IIA filaments drive cellular contractile system formation during interphase and mitosis

    PubMed Central

    Fenix, Aidan M.; Taneja, Nilay; Buttler, Carmen A.; Lewis, John; Van Engelenburg, Schuyler B.; Ohi, Ryoma; Burnette, Dylan T.

    2016-01-01

    Cell movement and cytokinesis are facilitated by contractile forces generated by the molecular motor, nonmuscle myosin II (NMII). NMII molecules form a filament (NMII-F) through interactions of their C-terminal rod domains, positioning groups of N-terminal motor domains on opposite sides. The NMII motors then bind and pull actin filaments toward the NMII-F, thus driving contraction. Inside of crawling cells, NMIIA-Fs form large macromolecular ensembles (i.e., NMIIA-F stacks), but how this occurs is unknown. Here we show NMIIA-F stacks are formed through two non–mutually exclusive mechanisms: expansion and concatenation. During expansion, NMIIA molecules within the NMIIA-F spread out concurrent with addition of new NMIIA molecules. Concatenation occurs when multiple NMIIA-Fs/NMIIA-F stacks move together and align. We found that NMIIA-F stack formation was regulated by both motor activity and the availability of surrounding actin filaments. Furthermore, our data showed expansion and concatenation also formed the contractile ring in dividing cells. Thus interphase and mitotic cells share similar mechanisms for creating large contractile units, and these are likely to underlie how other myosin II–based contractile systems are assembled. PMID:26960797

  2. Troponin-like regulation in muscle thin filaments of the mussel Crenomytilus grayanus (Bivalvia: Mytiloida).

    PubMed

    Vyatchin, Ilya G; Shevchenko, Ulyana V; Lazarev, Stanislav S; Matusovsky, Oleg S; Shelud'ko, Nikolay S

    2015-10-01

    Muscles of bivalve molluscs have double calcium regulation--myosin-linked and actin-linked. While the mechanism of myosin-linked regulation is sufficiently studied, there is still no consensus on the mechanism of actin-linked regulation. Earlier we showed a high degree of Ca2+-sensitivity of thin filaments from the adductor muscle of the mussel Crenomytilus grayanus (Mytiloida). In order to elucidate the nature of this regulation, we isolated the fraction of minor proteins from the mussel thin filaments, which confers Ca2+-sensitivity to reconstituted actomyosin-tropomyosin. Proteins of this fraction, ABP-19, ABP-20, and ABP-28, were chromatographically purified and identified. According to the results of mass spectrometry and Western blot analysis, as well as by their functional properties, these mussel actin-binding proteins appeared to correspond to the troponin components from the skeletal muscles of vertebrates (TnC, TnI and TnT). The reconstituted mussel troponin complex confers to actomyosin-tropomyosin more than 80% Ca2+-sensitivity. The in vivo molar ratio of actin/tropomyosin/troponin was calculated to be 7:1:0.5, i.e., the content of troponin in mussel thin filaments is two times lower than in thin filaments of skeletal muscles of vertebrates. These data demonstrate that troponin-like regulation found in the catch muscle of the mussel C. grayanus is present at least in two suborders of bivalves: Pectinoida and Mytiloida. Copyright © 2015 Elsevier B.V. All rights reserved.

  3. Tropomodulins: pointed-end capping proteins that regulate actin filament architecture in diverse cell types

    PubMed Central

    Yamashiro, Sawako; Gokhin, David S.; Kimura, Sumiko; Nowak, Roberta B.; Fowler, Velia M.

    2012-01-01

    Tropomodulins are a family of four proteins (Tmods 1–4) that cap the pointed ends of actin filaments in actin cytoskeletal structures in a developmentally regulated and tissue-specific manner. Unique among capping proteins, Tmods also bind tropomyosins (TMs), which greatly enhance the actin filament pointed-end capping activity of Tmods. Tmods are defined by a tropomyosin (TM)-regulated/Pointed-End Actin Capping (TM-Cap) domain in their unstructured N-terminal portion, followed by a compact, folded Leucine-Rich Repeat/Pointed-End Actin Capping (LRR-Cap) domain. By inhibiting actin monomer association and dissociation from pointed ends, Tmods regulate regulate actin dynamics and turnover, stabilizing actin filament lengths and cytoskeletal architecture. In this review, we summarize the genes, structural features, molecular and biochemical properties, actin regulatory mechanisms, expression patterns, and cell and tissue functions of Tmods. By understanding Tmods’ functions in the context of their molecular structure, actin regulation, binding partners, and related variants (leiomodins 1–3), we can draw broad conclusions that can explain the diverse morphological and functional phenotypes that arise from Tmod perturbation experiments in vitro and in vivo. Tmod-based stabilization and organization of intracellular actin filament networks provide key insights into how the emergent properties of the actin cytoskeleton drive tissue morphogenesis and physiology. PMID:22488942

  4. The role of keratin subfamilies and keratin pairs in the formation of human epidermal intermediate filaments

    PubMed Central

    1986-01-01

    The four major keratins of normal human epidermis (molecular mass 50, 56.5, 58, and 65-67 kD) can be subdivided on the basis of charge into two subfamilies (acidic 50-kD and 56.5-kD keratins vs. relatively basic 58-kD and 65-67-kD keratins) or subdivided on the basis of co- expression into two "pairs" (50-kD/58-kD keratin pair synthesized by basal cells vs. 56.5-kD/65-67-kD keratin pair expressed in suprabasal cells). Acidic and basic subfamilies were separated by ion exchange chromatography in 8.5 M urea and tested for their ability to reassemble into 10-nm filaments in vitro. The two keratins in either subfamily did not reassemble into 10-nm filaments unless combined with members of the other subfamily. While electron microscopy of acidic and basic keratins equilibrated in 4.5 M urea showed that keratins within each subfamily formed distinct oligomeric structures, possibly representing precursors in filament assembly, chemical cross-linking followed by gel analysis revealed dimers and larger oligomers only when subfamilies were combined. In addition, among the four major keratins, the acidic 50-kD and basic 58-kD keratins showed preferential association even in 8.5 M urea, enabling us to isolate a 50-kD/58-kD keratin complex by gel filtration. This isolated 50-kD/58-kD keratin pair readily formed 10-nm filaments in vitro. These results demonstrate that in tissues containing multiple keratins, two keratins are sufficient for filament assembly, but one keratin from each subfamily is required. More importantly, these data provide the first evidence for the structural significance of specific co-expressed acidic/basic keratin pairs in the formation of epithelial 10-nm filaments. PMID:2422179

  5. E93K charge reversal on actin perturbs steric regulation of thin filaments.

    PubMed

    Cammarato, Anthony; Craig, Roger; Sparrow, John C; Lehman, William

    2005-04-15

    Contraction in striated muscles is regulated by Ca2+-dependent movement of tropomyosin-troponin on thin filaments. Interactions of charged amino acid residues between the surfaces of tropomyosin and actin are believed to play an integral role in this steric mechanism by influencing the position of tropomyosin on the filaments. To investigate this possibility further, thin filaments were isolated from troponin-regulated, indirect flight muscles of Drosophila mutants that express actin with an amino acid charge reversal at residue 93 located at the interface between actin subdomains 1 and 2, in which a lysine residue is substituted for a glutamic acid. Electron microscopy and 3D helical reconstruction were employed to evaluate the structural effects of the mutation. In the absence of Ca2+, tropomyosin was in a position that blocked the myosin-binding sites on actin, as previously found with wild-type filaments. However, in the presence of Ca2+, tropomyosin position in the mutant filaments was much more variable than in the wild-type ones. In most cases (approximately 60%), tropomyosin remained in the blocking position despite the presence of Ca2+, failing to undergo a normal Ca2+-induced change in position. Thus, switching of a negative to a positive charge at position 93 on actin may stabilize negatively charged tropomyosin in the Ca2+-free state regardless of Ca2+ levels, an alteration that, in turn, is likely to interfere with steric regulation and consequently muscle activation. These results highlight the importance of actin's surface charges in determining the distribution of tropomyosin positions on thin filaments derived from troponin-regulated striated muscles.

  6. Invertebrate muscles: thin and thick filament structure; molecular basis of contraction and its regulation, catch and asynchronous muscle

    PubMed Central

    Hooper, Scott L.; Hobbs, Kevin H.; Thuma, Jeffrey B.

    2008-01-01

    This is the second in a series of canonical reviews on invertebrate muscle. We cover here thin and thick filament structure, the molecular basis of force generation and its regulation, and two special properties of some invertebrate muscle, catch and asynchronous muscle. Invertebrate thin filaments resemble vertebrate thin filaments, although helix structure and tropomyosin arrangement show small differences. Invertebrate thick filaments, alternatively, are very different from vertebrate striated thick filaments and show great variation within invertebrates. Part of this diversity stems from variation in paramyosin content, which is greatly increased in very large diameter invertebrate thick filaments. Other of it arises from relatively small changes in filament backbone structure, which results in filaments with grossly similar myosin head placements (rotating crowns of heads every 14.5 nm) but large changes in detail (distances between heads in azimuthal registration varying from three to thousands of crowns). The lever arm basis of force generation is common to both vetebrates and invertebrates, and in some invertebrates this process is understood on the near atomic level. Invertebrate actomyosin is both thin (tropomyosin:troponin) and thick (primarily via direct Ca++ binding to myosin) filament regulated, and most invertebrate muscles are dually regulated. These mechanisms are well understood on the molecular level, but the behavioral utility of dual regulation is less so. The phosphorylation state of the thick filament associated giant protein, twitchin, has been recently shown to be the molecular basis of catch. The molecular basis of the stretch activation underlying asynchronous muscle activity, however, remains unresolved. PMID:18616971

  7. Invertebrate muscles: thin and thick filament structure; molecular basis of contraction and its regulation, catch and asynchronous muscle.

    PubMed

    Hooper, Scott L; Hobbs, Kevin H; Thuma, Jeffrey B

    2008-10-01

    This is the second in a series of canonical reviews on invertebrate muscle. We cover here thin and thick filament structure, the molecular basis of force generation and its regulation, and two special properties of some invertebrate muscle, catch and asynchronous muscle. Invertebrate thin filaments resemble vertebrate thin filaments, although helix structure and tropomyosin arrangement show small differences. Invertebrate thick filaments, alternatively, are very different from vertebrate striated thick filaments and show great variation within invertebrates. Part of this diversity stems from variation in paramyosin content, which is greatly increased in very large diameter invertebrate thick filaments. Other of it arises from relatively small changes in filament backbone structure, which results in filaments with grossly similar myosin head placements (rotating crowns of heads every 14.5 nm) but large changes in detail (distances between heads in azimuthal registration varying from three to thousands of crowns). The lever arm basis of force generation is common to both vertebrates and invertebrates, and in some invertebrates this process is understood on the near atomic level. Invertebrate actomyosin is both thin (tropomyosin:troponin) and thick (primarily via direct Ca(++) binding to myosin) filament regulated, and most invertebrate muscles are dually regulated. These mechanisms are well understood on the molecular level, but the behavioral utility of dual regulation is less so. The phosphorylation state of the thick filament associated giant protein, twitchin, has been recently shown to be the molecular basis of catch. The molecular basis of the stretch activation underlying asynchronous muscle activity, however, remains unresolved.

  8. Filamentous Fungi in Drinking Water, Particularly in Relation to Biofilm Formation

    PubMed Central

    Siqueira, Virgínia M.; Oliveira, Helena M. B.; Santos, Cledir; Paterson, R. Russell M.; Gusmão, Norma B.; Lima, Nelson

    2011-01-01

    The presence of filamentous fungi in drinking water has become an area worthy of investigation with various studies now being published. The problems associated with fungi include blockage of water pipes, organoleptic deterioration, pathogenic fungi and mycotoxins. Fungal biofilm formation is a less developed field of study. This paper updates the topic and introduces novel methods on fungal biofilm analysis, particularly from work based in Brazil. Further recommendations for standard methodology are provided. PMID:21556196

  9. Three-color single molecule imaging shows WASP detachment from Arp2/3 complex triggers actin filament branch formation

    PubMed Central

    Smith, Benjamin A; Padrick, Shae B; Doolittle, Lynda K; Daugherty-Clarke, Karen; Corrêa, Ivan R; Xu, Ming-Qun; Goode, Bruce L; Rosen, Michael K; Gelles, Jeff

    2013-01-01

    During cell locomotion and endocytosis, membrane-tethered WASP proteins stimulate actin filament nucleation by the Arp2/3 complex. This process generates highly branched arrays of filaments that grow toward the membrane to which they are tethered, a conflict that seemingly would restrict filament growth. Using three-color single-molecule imaging in vitro we revealed how the dynamic associations of Arp2/3 complex with mother filament and WASP are temporally coordinated with initiation of daughter filament growth. We found that WASP proteins dissociated from filament-bound Arp2/3 complex prior to new filament growth. Further, mutations that accelerated release of WASP from filament-bound Arp2/3 complex proportionally accelerated branch formation. These data suggest that while WASP promotes formation of pre-nucleation complexes, filament growth cannot occur until it is triggered by WASP release. This provides a mechanism by which membrane-bound WASP proteins can stimulate network growth without restraining it. DOI: http://dx.doi.org/10.7554/eLife.01008.001 PMID:24015360

  10. Septin9 is involved in septin filament formation and cellular stability.

    PubMed

    Füchtbauer, Annette; Lassen, Louise B; Jensen, Astrid B; Howard, Jennifer; Quiroga, Adán de Salas; Warming, Søren; Sørensen, Annette B; Pedersen, Finn S; Füchtbauer, Ernst-Martin

    2011-08-01

    Septin9 (Sept9) is a member of the filament-forming septin family of structural proteins and is associated with a variety of cancers and with hereditary neuralgic amyotrophy. We have generated mice with constitutive and conditional Sept9 knockout alleles. Homozygous deletion of Sept9 results in embryonic lethality around day 10 of gestation whereas mice homozygous for the conditional allele develop normally. Here we report the consequences of homozygous loss of Sept9 in immortalized murine embryonic fibroblasts. Proliferation rate was not changed but cells without Sept9 had an altered morphology compared to normal cells, particularly under low serum stress. Abnormal, fragmented, and multiple nuclei were more frequent in cells without Sept9. Cell migration, as measured by gap-filling and filter-invasion assays, was impaired, but individual cells did not move less than wild-type cells. Sept9 knockout cells showed a reduced resistance to hypo-osmotic stress. Stress fiber and vinculin staining at focal adhesion points was less prominent. Long septin filaments stained for Sept7 disappeared. Instead, staining was found in short, often curved filaments and rings. Furthermore, Sept7 was no longer localized to the mitotic spindle. Together, these data reveal the importance of Sept9 for septin filament formation and general cell stability.

  11. Filament formation associated with spirochetal infection: a comparative approach to Morgellons disease.

    PubMed

    Middelveen, Marianne J; Stricker, Raphael B

    2011-01-01

    Bovine digital dermatitis is an emerging infectious disease that causes lameness, decreased milk production, and weight loss in livestock. Proliferative stages of bovine digital dermatitis demonstrate keratin filament formation in skin above the hooves in affected animals. The multifactorial etiology of digital dermatitis is not well understood, but spirochetes and other coinfecting microorganisms have been implicated in the pathogenesis of this veterinary illness. Morgellons disease is an emerging human dermopathy characterized by the presence of filamentous fibers of undetermined composition, both in lesions and subdermally. While the etiology of Morgellons disease is unknown, there is serological and clinical evidence linking this phenomenon to Lyme borreliosis and coinfecting tick-borne agents. Although the microscopy of Morgellons filaments has been described in the medical literature, the structure and pathogenesis of these fibers is poorly understood. In contrast, most microscopy of digital dermatitis has focused on associated pathogens and histology rather than the morphology of late-stage filamentous fibers. Clinical, laboratory, and microscopic characteristics of these two diseases are compared.

  12. Filament formation associated with spirochetal infection: a comparative approach to Morgellons disease

    PubMed Central

    Middelveen, Marianne J; Stricker, Raphael B

    2011-01-01

    Bovine digital dermatitis is an emerging infectious disease that causes lameness, decreased milk production, and weight loss in livestock. Proliferative stages of bovine digital dermatitis demonstrate keratin filament formation in skin above the hooves in affected animals. The multifactorial etiology of digital dermatitis is not well understood, but spirochetes and other coinfecting microorganisms have been implicated in the pathogenesis of this veterinary illness. Morgellons disease is an emerging human dermopathy characterized by the presence of filamentous fibers of undetermined composition, both in lesions and subdermally. While the etiology of Morgellons disease is unknown, there is serological and clinical evidence linking this phenomenon to Lyme borreliosis and coinfecting tick-borne agents. Although the microscopy of Morgellons filaments has been described in the medical literature, the structure and pathogenesis of these fibers is poorly understood. In contrast, most microscopy of digital dermatitis has focused on associated pathogens and histology rather than the morphology of late-stage filamentous fibers. Clinical, laboratory, and microscopic characteristics of these two diseases are compared. PMID:22253541

  13. Cluster-formation in the Rosette molecular cloud at the junctions of filaments

    NASA Astrophysics Data System (ADS)

    Schneider, N.; Csengeri, T.; Hennemann, M.; Motte, F.; Didelon, P.; Federrath, C.; Bontemps, S.; Di Francesco, J.; Arzoumanian, D.; Minier, V.; André, Ph.; Hill, T.; Zavagno, A.; Nguyen-Luong, Q.; Attard, M.; Bernard, J.-Ph.; Elia, D.; Fallscheer, C.; Griffin, M.; Kirk, J.; Klessen, R.; Könyves, V.; Martin, P.; Men'shchikov, A.; Palmeirim, P.; Peretto, N.; Pestalozzi, M.; Russeil, D.; Sadavoy, S.; Sousbie, T.; Testi, L.; Tremblin, P.; Ward-Thompson, D.; White, G.

    2012-04-01

    Aims: For many years feedback processes generated by OB-stars in molecular clouds, including expanding ionization fronts, stellar winds, or UV-radiation, have been proposed to trigger subsequent star formation. However, hydrodynamic models including radiation and gravity show that UV-illumination has little or no impact on the global dynamical evolution of the cloud. Instead, gravitational collapse of filaments and/or merging of filamentary structures can lead to building up dense high-mass star-forming clumps. However, the overall density structure of the cloud has a large influence on this process, and requires a better understanding. Methods: The Rosette molecular cloud, irradiated by the NGC 2244 cluster, is a template region for triggered star-formation, and we investigated its spatial and density structure by applying a curvelet analysis, a filament-tracing algorithm (DisPerSE), and probability density functions (PDFs) on Herschel column density maps, obtained within the HOBYS key program. Results: The analysis reveals not only the filamentary structure of the cloud but also that all known infrared clusters except one lie at junctions of filaments, as predicted by turbulence simulations. The PDFs of sub-regions in the cloud show systematic differences. The two UV-exposed regions have a double-peaked PDF we interprete as caused by shock compression, while the PDFs of the center and other cloud parts are more complex, partly with a power-law tail. A deviation of the log-normal PDF form occurs at AV ≈ 9m for the center, and around 4m for the other regions. Only the part of the cloud farthest from the Rosette nebula shows a log-normal PDF. Conclusions: The deviations of the PDF from the log-normal shape typically associated with low- and high-mass star-forming regions at AV ≈ 3-4m and 8-10m, respectively, are found here within the very same cloud. This shows that there is no fundamental difference in the density structure of low- and high-mass star

  14. Different inhibitory response of cyanidin and methylene blue for filament formation of tau microtubule-binding domain

    SciTech Connect

    Hattori, Masaki; Sugino, Etsuko; Minoura, Katsuhiko In, Yasuko; Sumida, Miho; Taniguchi, Taizo; Tomoo, Koji; Ishida, Toshimasa

    2008-09-12

    One of the priorities in Alzheimer research is to develop a compound that inhibits the filament formation of tau protein. Since the three- or four-repeat microtubule-binding domain (MBD) in tau protein plays an essential role in filament formation, the inhibitory behavior of cyanidin (Cy) and methylene blue (MB) with respect to heparin-induced filament formation of MBD in a neutral solution (pH 7.6) was characterized by fluorescence, circular dichroism, and electron microscopy measurements. The planar aromatic ring of Cy and the N-unsubstituted phenothiazine ring of MB were shown to be necessary for the inhibition. However, the inhibitory responses with respect to heparin-induced filament formation to the second and third repeat peptides of MBD were different: Cy suppresses the formation and MB does not prevent the formation. This suggests the importance of the first and fourth repeat peptides in the inhibitory activity of MB for MBD filament formation. In this study, we showed that the decrease of thioflavin S fluorescence intensity is not always linked to inhibition of filament formation.

  15. Barrier role of actin filaments in regulated mucin secretion from airway goblet cells.

    PubMed

    Ehre, Camille; Rossi, Andrea H; Abdullah, Lubna H; De Pestel, Kathleen; Hill, Sandra; Olsen, John C; Davis, C William

    2005-01-01

    Airway goblet cells secrete mucin onto mucosal surfaces under the regulation of an apical, phospholipase C/G(q)-coupled P2Y(2) receptor. We tested whether cortical actin filaments negatively regulate exocytosis in goblet cells by forming a barrier between secretory granules and plasma membrane docking sites as postulated for other secretory cells. Immunostaining of human lung tissues and SPOC1 cells (an epithelial, mucin-secreting cell line) revealed an apical distribution of beta- and gamma-actin in ciliated and goblet cells. In goblet cells, actin appeared as a prominent subplasmalemmal sheet lying between granules and the apical membrane, and it disappeared from SPOC1 cells activated by purinergic agonist. Disruption of actin filaments with latrunculin A stimulated SPOC1 cell mucin secretion under basal and agonist-activated conditions, whereas stabilization with jasplakinolide or overexpression of beta- or gamma-actin conjugated to yellow fluorescent protein (YFP) inhibited secretion. Myristoylated alanine-rich C kinase substrate, a PKC-activated actin-plasma membrane tethering protein, was phosphorylated after agonist stimulation, suggesting a translocation to the cytosol. Scinderin (or adseverin), a Ca(2+)-activated actin filament severing and capping protein was cloned from human airway and SPOC1 cells, and synthetic peptides corresponding to its actin-binding domains inhibited mucin secretion. We conclude that actin filaments negatively regulate mucin secretion basally in airway goblet cells and are dynamically remodeled in agonist-stimulated cells to promote exocytosis.

  16. Biofilm formation in Desulfovibrio vulgaris Hildenborough is dependent upon protein filaments.

    PubMed

    Clark, Melinda E; Edelmann, Richard E; Duley, Matt L; Wall, Judy D; Fields, Matthew W

    2007-11-01

    Desulfovibrio vulgaris Hildenborough is a Gram-negative sulfate-reducing bacterium (SRB), and the physiology of SRBs can impact many anaerobic environments including radionuclide waste sites, oil reservoirs and metal pipelines. In an attempt to understand D. vulgaris as a population that can adhere to surfaces, D. vulgaris cultures were grown in a defined medium and analysed for carbohydrate production, motility and biofilm formation. Desulfovibrio vulgaris wild-type cells had increasing amounts of carbohydrate into stationary phase and approximately half of the carbohydrate remained internal. In comparison, a mutant that lacked the 200 kb megaplasmid, strain DeltaMP, produced less carbohydrate and the majority of carbohydrate remained internal of the cell proper. To assess the possibility of carbohydrate re-allocation, biofilm formation was investigated. Wild-type cells produced approximately threefold more biofilm on glass slides compared with DeltaMP; however, wild-type biofilm did not contain significant levels of exopolysaccharide. In addition, stains specific for extracellular carbohydrate did not reveal polysaccharide material within the biofilm. Desulfovibrio vulgaris wild-type biofilms contained long filaments as observed with scanning electron microscopy (SEM), and the biofilm-deficient DeltaMP strain was also deficient in motility. Biofilms grown directly on silica oxide transmission electron microscopy (TEM) grids did not contain significant levels of an exopolysaccharide matrix when viewed with TEM and SEM, and samples stained with ammonium molybdate also showed long filaments that resembled flagella. Biofilms subjected to protease treatments were degraded, and different proteases that were added at the time of inoculation inhibited biofilm formation. The data indicated that D. vulgaris did not produce an extensive exopolysaccharide matrix, used protein filaments to form biofilm between cells and silica oxide surfaces, and the filaments appeared to be

  17. Divalent cations crosslink vimentin intermediate filament tail domains to regulate network mechanics.

    PubMed

    Lin, Yi-Chia; Broedersz, Chase P; Rowat, Amy C; Wedig, Tatjana; Herrmann, Harald; Mackintosh, Frederick C; Weitz, David A

    2010-06-18

    Intermediate filament networks in the cytoplasm and nucleus are critical for the mechanical integrity of metazoan cells. However, the mechanism of crosslinking in these networks and the origins of their mechanical properties are not understood. Here, we study the elastic behavior of in vitro networks of the intermediate filament protein vimentin. Rheological experiments reveal that vimentin networks stiffen with increasing concentrations of Ca(2+) and Mg(2+), showing that divalent cations act as crosslinkers. We quantitatively describe the elastic response of vimentin networks over five decades of applied stress using a theory that treats the divalent cations as crosslinkers: at low stress, the behavior is entropic in origin, and increasing stress pulls out thermal fluctuations from single filaments, giving rise to a nonlinear response; at high stress, enthalpic stretching of individual filaments significantly modifies the nonlinearity. We investigate the elastic properties of networks formed by a series of protein variants with stepwise tail truncations and find that the last 11 amino acids of the C-terminal tail domain mediate crosslinking by divalent ions. We determined the single-filament persistence length, l(P) approximately 0.5 mum, and Young's modulus, Y approximately 9 MPa; both are consistent with literature values. Our results provide insight into a crosslinking mechanism for vimentin networks and suggest that divalent ions may help regulate the cytoskeletal structure and mechanical properties of cells. Copyright (c) 2010 Elsevier Ltd. All rights reserved.

  18. An Integrative Approach for Modeling and Simulation of Heterocyst Pattern Formation in Cyanobacteria Filaments

    PubMed Central

    Torres-Sánchez, Alejandro; Gómez-Gardeñes, Jesús; Falo, Fernando

    2015-01-01

    Heterocyst differentiation in cyanobacteria filaments is one of the simplest examples of cellular differentiation and pattern formation in multicellular organisms. Despite of the many experimental studies addressing the evolution and sustainment of heterocyst patterns and the knowledge of the genetic circuit underlying the behavior of single cyanobacterium under nitrogen deprivation, there is still a theoretical gap connecting these two macroscopic and microscopic processes. As an attempt to shed light on this issue, here we explore heterocyst differentiation under the paradigm of systems biology. This framework allows us to formulate the essential dynamical ingredients of the genetic circuit of a single cyanobacterium into a set of differential equations describing the time evolution of the concentrations of the relevant molecular products. As a result, we are able to study the behavior of a single cyanobacterium under different external conditions, emulating nitrogen deprivation, and simulate the dynamics of cyanobacteria filaments by coupling their respective genetic circuits via molecular diffusion. These two ingredients allow us to understand the principles by which heterocyst patterns can be generated and sustained. In particular, our results point out that, by including both diffusion and noisy external conditions in the computational model, it is possible to reproduce the main features of the formation and sustainment of heterocyst patterns in cyanobacteria filaments as observed experimentally. Finally, we discuss the validity and possible improvements of the model. PMID:25816286

  19. The antagonistic effect of Saccharomyces boulardii on Candida albicans filamentation, adhesion and biofilm formation.

    PubMed

    Krasowska, Anna; Murzyn, Anna; Dyjankiewicz, Agnieszka; Łukaszewicz, Marcin; Dziadkowiec, Dorota

    2009-12-01

    The dimorphic fungus Candida albicans is a member of the normal flora residing in the intestinal tract of humans. In spite of this, under certain conditions it can induce both superficial and serious systemic diseases, as well as be the cause of gastrointestinal infections. Saccharomyces boulardii is a yeast strain that has been shown to have applications in the prevention and treatment of intestinal infections caused by bacterial pathogens. The purpose of this study was to determine whether S. boulardii affects the virulence factors of C. albicans. We demonstrate the inhibitory effect of live S. boulardii cells on the filamentation (hyphae and pseudohyphae formation) of C. albicans SC5314 strain proportional to the amount of S. boulardii added. An extract from S. boulardii culture has a similar effect. Live S. boulardii and the extract from S. boulardii culture filtrate diminish C. albicans adhesion to and subsequent biofilm formation on polystyrene surfaces under both aerobic and microaerophilic conditions. This effect is very strong and requires lower doses of S. boulardii cells or concentrations of the extract than serum-induced filamentation tests. Saccharomyces boulardii has a strong negative effect on very important virulence factors of C. albicans, i.e. the ability to form filaments and to adhere and form biofilms on plastic surfaces.

  20. Filaments in curved streamlines: Rapid formation of Staphylococcus aureus biofilm streamers

    PubMed Central

    Kim, Minyoung Kevin; Drescher, Knut; Pak, On Shun; Bassler, Bonnie L.; Stone, Howard A.

    2014-01-01

    Biofilms are surface-associated conglomerates of bacteria that are highly resistant to antibiotics. These bacterial communities can cause chronic infections in humans by colonizing, for example, medical implants, heart valves, or lungs. Staphylococcus aureus, a notorious human pathogen, causes some of the most common biofilm-related infections. Despite the clinical importance of S. aureus biofilms, it remains mostly unknown how physical effects, in particular flow, and surface structure influence biofilm dynamics. Here we use model microfluidic systems to investigate how environmental factors, such as surface geometry, surface chemistry, and fluid flow affect biofilm development in S. aureus. We discovered that S. aureus rapidly forms flow-induced, filamentous biofilm streamers, and furthermore if surfaces are coated with human blood plasma, streamers appear within minutes and clog the channels more rapidly than if the channels are uncoated. To understand how biofilm streamer filaments reorient in flows with curved streamlines to bridge the distances between corners, we developed a mathematical model based on resistive force theory of slender filaments. Understanding physical aspects of biofilm formation in S. aureus may lead to new approaches for interrupting biofilm formation of this pathogen. PMID:25484614

  1. Filaments in curved flow: Rapid formation of Staphylococcus aureus biofilm streamers

    NASA Astrophysics Data System (ADS)

    Kim, Min Young; Drescher, Knut; Pak, On Shun; Bassler, Bonnie L.; Stone, Howard A.

    2014-03-01

    Biofilms are surface-associated conglomerates of bacteria that are highly resistant to antibiotics. These bacterial communities can cause chronic infections in humans by colonizing, for example, medical implants, heart valves, or lungs. Staphylococcus aureus, a notorious human pathogen, causes some of the most common biofilm-related infections. Despite the clinical importance of S. aureus biofilms, it remains mostly unknown how physical effects, in particular flow, and surface structure influence biofilm dynamics. Here we use model microfluidic systems to investigate how environmental factors, such as surface geometry, surface chemistry, and fluid flow affect biofilm development in S. aureus.We discovered that S. aureus rapidly forms flow-induced, filamentous biofilm streamers, and furthermore if surfaces are coated with human blood plasma, streamers appear within minutes and clog the channels more rapidly than if the channels are uncoated. To understand how biofilm streamer filaments reorient in curved flow to bridge the distances between corners, we developed a mathematical model based on resistive force theory and slender filaments. Understanding physical aspects of biofilm formation in S. aureus may lead to new approaches for interrupting biofilm formation of this pathogen.

  2. Filaments in curved streamlines: rapid formation of Staphylococcus aureus biofilm streamers

    NASA Astrophysics Data System (ADS)

    Kim, Minyoung Kevin; Drescher, Knut; Pak, On Shun; Bassler, Bonnie L.; Stone, Howard A.

    2014-06-01

    Biofilms are surface-associated conglomerates of bacteria that are highly resistant to antibiotics. These bacterial communities can cause chronic infections in humans by colonizing, for example, medical implants, heart valves, or lungs. Staphylococcus aureus, a notorious human pathogen, causes some of the most common biofilm-related infections. Despite the clinical importance of S. aureus biofilms, it remains mostly unknown how physical effects, in particular flow, and surface structure influence biofilm dynamics. Here we use model microfluidic systems to investigate how environmental factors, such as surface geometry, surface chemistry, and fluid flow affect biofilm development of S. aureus. We discovered that S. aureus rapidly forms flow-induced, filamentous biofilm streamers, and furthermore if surfaces are coated with human blood plasma, streamers appear within minutes and clog the channels more rapidly than if the channels are uncoated. To understand how biofilm streamer filaments reorient in flows with curved streamlines to bridge the distances between corners, we developed a mathematical model based on resistive force theory of slender filaments. Understanding physical aspects of biofilm formation of S. aureus may lead to new approaches for interrupting biofilm formation of this pathogen.

  3. Role of Reactive Intermediates in Manganese Oxide Formation By Filamentous Ascomycete Fungi

    NASA Astrophysics Data System (ADS)

    Zeiner, C. A.; Anderton, C.; Wu, S.; Purvine, S.; Zink, E.; Paša-Tolić, L.; Santelli, C. M.; Hansel, C. M.

    2014-12-01

    Biogenic manganese (Mn) oxide minerals are ubiquitous in the environment, and their high reactivity can profoundly impact the fate of contaminants and cycling of carbon and nutrients. In contrast to bacteria, the pathways utilized by fungi to oxidize Mn(II) to Mn(III,IV) oxides remain largely unknown. Here, we explore the mechanisms of Mn(II) oxidation by a phylogenetically diverse group of filamentous Ascomycete fungi using a combination of chemical assays and bulk and spatially-resolved mass spectrometry. We show that the mechanisms of Mn(II) oxidation vary with fungal species, over time during secretome compositional changes, and in the presence of other fungi. Specifically, our work implicates a dynamic transition in Mn(II) oxidation pathways that varies between species. In particular, while reactive oxygen species (ROS) produced via transmembrane NADPH oxidases are involved in initial oxidation, over time, secreted enzymes become important Mn(II) oxidation mediators for some species. In addition, the overall secretome oxidation capacity varies with time and fungal species. Secretome analysis reveals a surprising absence of enzymes currently considered to be Mn(II)-oxidizing enzymes in these organisms, and instead highlights a wide variety of redox-active enzymes. Furthermore, we implicate fungal cell defense mechanisms in the formation of distinct Mn oxide patterns when fungi are grown in head-to-head competition. The identification and regulation of these secreted enzymes are under current investigation within the bulk secretome and within the interaction zone of structured fungal communities. Overall, our findings illustrate that Ascomycete Mn(II) oxidation mechanisms are highly variable and are dictated by complex environmental and ecological interactions. Future work will explore the connection between Ascomycete Mn(II) oxidation and the ability to degrade cellulose, a key carbon reservoir for biofuel production.

  4. ER sheet persistence is coupled to myosin 1c-regulated dynamic actin filament arrays.

    PubMed

    Joensuu, Merja; Belevich, Ilya; Rämö, Olli; Nevzorov, Ilya; Vihinen, Helena; Puhka, Maija; Witkos, Tomasz M; Lowe, Martin; Vartiainen, Maria K; Jokitalo, Eija

    2014-04-01

    The endoplasmic reticulum (ER) comprises a dynamic three-dimensional (3D) network with diverse structural and functional domains. Proper ER operation requires an intricate balance within and between dynamics, morphology, and functions, but how these processes are coupled in cells has been unclear. Using live-cell imaging and 3D electron microscopy, we identify a specific subset of actin filaments localizing to polygons defined by ER sheets and tubules and describe a role for these actin arrays in ER sheet persistence and, thereby, in maintenance of the characteristic network architecture by showing that actin depolymerization leads to increased sheet fluctuation and transformations and results in small and less abundant sheet remnants and a defective ER network distribution. Furthermore, we identify myosin 1c localizing to the ER-associated actin filament arrays and reveal a novel role for myosin 1c in regulating these actin structures, as myosin 1c manipulations lead to loss of the actin filaments and to similar ER phenotype as observed after actin depolymerization. We propose that ER-associated actin filaments have a role in ER sheet persistence regulation and thus support the maintenance of sheets as a stationary subdomain of the dynamic ER network.

  5. ER sheet persistence is coupled to myosin 1c–regulated dynamic actin filament arrays

    PubMed Central

    Joensuu, Merja; Belevich, Ilya; Rämö, Olli; Nevzorov, Ilya; Vihinen, Helena; Puhka, Maija; Witkos, Tomasz M.; Lowe, Martin; Vartiainen, Maria K.; Jokitalo, Eija

    2014-01-01

    The endoplasmic reticulum (ER) comprises a dynamic three-dimensional (3D) network with diverse structural and functional domains. Proper ER operation requires an intricate balance within and between dynamics, morphology, and functions, but how these processes are coupled in cells has been unclear. Using live-cell imaging and 3D electron microscopy, we identify a specific subset of actin filaments localizing to polygons defined by ER sheets and tubules and describe a role for these actin arrays in ER sheet persistence and, thereby, in maintenance of the characteristic network architecture by showing that actin depolymerization leads to increased sheet fluctuation and transformations and results in small and less abundant sheet remnants and a defective ER network distribution. Furthermore, we identify myosin 1c localizing to the ER-associated actin filament arrays and reveal a novel role for myosin 1c in regulating these actin structures, as myosin 1c manipulations lead to loss of the actin filaments and to similar ER phenotype as observed after actin depolymerization. We propose that ER-associated actin filaments have a role in ER sheet persistence regulation and thus support the maintenance of sheets as a stationary subdomain of the dynamic ER network. PMID:24523293

  6. The Actin Filament-Binding Protein Coronin Regulates Motility in Plasmodium Sporozoites

    PubMed Central

    Bane, Kartik S.; Singer, Mirko; Reinig, Miriam; Klug, Dennis; Heiss, Kirsten; Baum, Jake; Mueller, Ann-Kristin; Frischknecht, Friedrich

    2016-01-01

    Parasites causing malaria need to migrate in order to penetrate tissue barriers and enter host cells. Here we show that the actin filament-binding protein coronin regulates gliding motility in Plasmodium berghei sporozoites, the highly motile forms of a rodent malaria-causing parasite transmitted by mosquitoes. Parasites lacking coronin show motility defects that impair colonization of the mosquito salivary glands but not migration in the skin, yet result in decreased transmission efficiency. In non-motile sporozoites low calcium concentrations mediate actin-independent coronin localization to the periphery. Engagement of extracellular ligands triggers an intracellular calcium release followed by the actin-dependent relocalization of coronin to the rear and initiation of motility. Mutational analysis and imaging suggest that coronin organizes actin filaments for productive motility. Using coronin-mCherry as a marker for the presence of actin filaments we found that protein kinase A contributes to actin filament disassembly. We finally speculate that calcium and cAMP-mediated signaling regulate a switch from rapid parasite motility to host cell invasion by differentially influencing actin dynamics. PMID:27409081

  7. Translation elongation factor EF-Tu modulates filament formation of actin-like MreB protein in vitro.

    PubMed

    Defeu Soufo, Hervé Joël; Reimold, Christian; Breddermann, Hannes; Mannherz, Hans G; Graumann, Peter L

    2015-04-24

    EF-Tu has been shown to interact with actin-like protein MreB and to affect its localization in Escherichia coli and in Bacillus subtilis cells. We have purified YFP-MreB in an active form, which forms filaments on glass slides in vitro and was active in dynamic light-scattering assays, polymerizing in milliseconds after addition of magnesium. Purified EF-Tu enhanced the amount of MreB filaments, as seen by sedimentation assays, the speed of filament formation and the length of MreB filaments in vitro. EF-Tu had the strongest impact on MreB filaments in a 1:1 ratio, and EF-Tu co-sedimented with MreB filaments, revealing a stoichiometric interaction between both proteins. This was supported by cross-linking assays where 1:1 species were well detectable. When expressed in E. coli cells, B. subtilis MreB formed filaments and induced the formation of co-localizing B. subtilis EF-Tu structures, indicating that MreB can direct the positioning of EF-Tu structures in a heterologous cell system. Fluorescence recovery after photobleaching analysis showed that MreB filaments have a higher turnover in B. subtilis cells than in E. coli cells, indicating different filament kinetics in homologous or heterologous cell systems. The data show that MreB can direct the localization of EF-Tu in vivo, which in turn positively affects the formation and dynamics of MreB filaments. Thus, EF-Tu is a modulator of the activity of a bacterial actin-like protein.

  8. Taxol-stabilized microtubules promote the formation of filaments from unmodified full-length Tau in vitro.

    PubMed

    Duan, Aranda R; Goodson, Holly V

    2012-12-01

    Tau is a neuronal protein that stabilizes the microtubule (MT) network, but it also forms filaments associated with Alzheimer's disease. Understanding Tau-MT and Tau-Tau interactions would help to establish Tau function in health and disease. For many years, literature reports on Tau-MT binding behavior and affinity have remained surprisingly contradictory (e.g., 10-fold variation in Tau-MT affinity). Tau-Tau interactions have also been investigated, but whether MTs might affect Tau filament formation is unknown. We have addressed these issues through binding assays and microscopy. We assessed Tau-MT interactions via cosedimentation and found that the measured affinity of Tau varies greatly, depending on the experimental design and the protein concentrations used. To investigate this dependence, we used fluorescence microscopy to examine Tau-MT binding. Strikingly, we found that Taxol-stabilized MTs promote Tau filament formation without characterized Tau-filament inducers. We propose that these novel Tau filaments account for the incongruence in Tau-MT affinity measurements. Moreover, electron microscopy reveals that these filaments appear similar to the heparin-induced Alzheimer's model. These observations suggest that the MT-induced Tau filaments provide a new model for Alzheimer's studies and that MTs might play a role in the formation of Alzheimer's-associated neurofibrillary tangles.

  9. THE FORMATION AND ERUPTION OF A SMALL CIRCULAR FILAMENT DRIVEN BY ROTATING MAGNETIC STRUCTURES IN THE QUIET SUN

    SciTech Connect

    Yang, Bo; Jiang, Yunchun; Yang, Jiayan; Hong, Junchao; Xu, Zhe E-mail: yjy@ynao.ac.cn

    2015-04-20

    We present the first observation of the formation and eruption of a small circular filament driven by a rotating network magnetic field (RNF) in the quiet Sun. In the negative footpoint region of an inverse J-shaped dextral filament, the RNF was formed by the convergence to supergranular junctions of several magnetic flux patches of the same polarity, and it then rotated counterclockwise (CCW) for approximately 11 hr and showed up as a CCW rotating EUV cyclone, during which time the filament gradually evolved into a circular filament that surrounded the cyclone. When the calculated convergence and vortex flows appeared around the RNF during its formation and rotation phases, the injected magnetic helicity calculation also showed negative helicity accumulation during the RNF rotation that was consistent with the dextral chirality of the filament. Finally, the RNF rotation stopped and the cyclone disappeared, and, probably due to an emerging bipole and its forced cancellation with the RNF, the closure filament underwent an eruption along its axis in the (clockwise) direction opposite to the rotation directions of the RNF and cyclone. These observations suggest that the RNFs might play an important role in the formation of nearby small-scale circular filaments as they transport and inject magnetic energy and helicity, and the formation of the EUV cyclones may be a further manifestation of the helicity injected into the corona by the rotation of the RNFs in the photosphere. In addition, the new emerging bipole observed before the filament eruption might be responsible for destabilizing the system and triggering the magnetic reconnection which proves useful for the filament eruption.

  10. The Formation and Eruption of a Small Circular Filament Driven by Rotating Magnetic Structures in the Quiet Sun

    NASA Astrophysics Data System (ADS)

    Yang, Bo; Jiang, Yunchun; Yang, Jiayan; Hong, Junchao; Xu, Zhe

    2015-04-01

    We present the first observation of the formation and eruption of a small circular filament driven by a rotating network magnetic field (RNF) in the quiet Sun. In the negative footpoint region of an inverse J-shaped dextral filament, the RNF was formed by the convergence to supergranular junctions of several magnetic flux patches of the same polarity, and it then rotated counterclockwise (CCW) for approximately 11 hr and showed up as a CCW rotating EUV cyclone, during which time the filament gradually evolved into a circular filament that surrounded the cyclone. When the calculated convergence and vortex flows appeared around the RNF during its formation and rotation phases, the injected magnetic helicity calculation also showed negative helicity accumulation during the RNF rotation that was consistent with the dextral chirality of the filament. Finally, the RNF rotation stopped and the cyclone disappeared, and, probably due to an emerging bipole and its forced cancellation with the RNF, the closure filament underwent an eruption along its axis in the (clockwise) direction opposite to the rotation directions of the RNF and cyclone. These observations suggest that the RNFs might play an important role in the formation of nearby small-scale circular filaments as they transport and inject magnetic energy and helicity, and the formation of the EUV cyclones may be a further manifestation of the helicity injected into the corona by the rotation of the RNFs in the photosphere. In addition, the new emerging bipole observed before the filament eruption might be responsible for destabilizing the system and triggering the magnetic reconnection which proves useful for the filament eruption.

  11. MTL–Independent Phenotypic Switching in Candida tropicalis and a Dual Role for Wor1 in Regulating Switching and Filamentation

    PubMed Central

    Porman, Allison M.; Wang, Na; Bennett, Richard J.

    2013-01-01

    Phenotypic switching allows for rapid transitions between alternative cell states and is important in pathogenic fungi for colonization and infection of different host niches. In Candida albicans, the white-opaque phenotypic switch plays a central role in regulating the program of sexual mating as well as interactions with the mammalian host. White-opaque switching is controlled by genes encoded at the MTL (mating-type-like) locus that ensures that only a or α cells can switch from the white state to the mating-competent opaque state, while a/α cells are refractory to switching. Here, we show that the related pathogen C. tropicalis undergoes white-opaque switching in all three cell types (a, α, and a/α), and thus switching is independent of MTL control. We also demonstrate that C. tropicalis white cells are themselves mating-competent, albeit at a lower efficiency than opaque cells. Transcriptional profiling of C. tropicalis white and opaque cells reveals significant overlap between switch-regulated genes in MTL homozygous and MTL heterozygous cells, although twice as many genes are white-opaque regulated in a/α cells as in a cells. In C. albicans, the transcription factor Wor1 is the master regulator of the white-opaque switch, and we show that Wor1 also regulates switching in C. tropicalis; deletion of WOR1 locks a, α, and a/α cells in the white state, while WOR1 overexpression induces these cells to adopt the opaque state. Furthermore, we show that WOR1 overexpression promotes both filamentous growth and biofilm formation in C. tropicalis, independent of the white-opaque switch. These results demonstrate an expanded role for C. tropicalis Wor1, including the regulation of processes necessary for infection of the mammalian host. We discuss these findings in light of the ancestral role of Wor1 as a transcriptional regulator of the transition between yeast form and filamentous growth. PMID:23555286

  12. Single-filament kinetic studies provide novel insights into regulation of actin-based motility

    PubMed Central

    Shekhar, Shashank; Carlier, Marie-France

    2016-01-01

    Polarized assembly of actin filaments forms the basis of actin-based motility and is regulated both spatially and temporally. Cells use a variety of mechanisms by which intrinsically slower processes are accelerated, and faster ones decelerated, to match rates observed in vivo. Here we discuss how kinetic studies of individual reactions and cycles that drive actin remodeling have provided a mechanistic and quantitative understanding of such processes. We specifically consider key barbed-end regulators such as capping protein and formins as illustrative examples. We compare and contrast different kinetic approaches, such as the traditional pyrene-polymerization bulk assays, as well as more recently developed single-filament and single-molecule imaging approaches. Recent development of novel biophysical methods for sensing and applying forces will in future allow us to address the very important relationship between mechanical stimulus and kinetics of actin-based motility. PMID:26715420

  13. Structural basis of the relaxed state of a Ca2+-regulated myosin filament and its evolutionary implications.

    PubMed

    Woodhead, John L; Zhao, Fa-Qing; Craig, Roger

    2013-05-21

    Myosin filaments of muscle are regulated either by phosphorylation of their regulatory light chains or Ca(2+) binding to the essential light chains, contributing to on-off switching or modulation of contraction. Phosphorylation-regulated filaments in the relaxed state are characterized by an asymmetric interaction between the two myosin heads, inhibiting their actin binding or ATPase activity. Here, we have tested whether a similar interaction switches off activity in myosin filaments regulated by Ca(2+) binding. Cryo-electron microscopy and single-particle image reconstruction of Ca(2+)-regulated (scallop) filaments reveals a helical array of myosin head-pair motifs above the filament surface. Docking of atomic models of scallop myosin head domains into the motifs reveals that the heads interact in a similar way to those in phosphorylation-regulated filaments. The results imply that the two major evolutionary branches of myosin regulation--involving phosphorylation or Ca(2+) binding--share a common structural mechanism for switching off thick-filament activity in relaxed muscle. We suggest that the Ca(2+)-binding mechanism evolved from the more ancient phosphorylation-based system to enable rapid response of myosin-regulated muscles to activation. Although the motifs are similar in both systems, the scallop structure is more tilted and higher above the filament backbone, leading to different intermolecular interactions. The reconstruction reveals how the myosin tail emerges from the motif, connecting the heads to the filament backbone, and shows that the backbone is built from supramolecular assemblies of myosin tails. The reconstruction provides a native structural context for understanding past biochemical and biophysical studies of this model Ca(2+)-regulated myosin.

  14. Auxin Regulates Arabidopsis Anther Dehiscence, Pollen Maturation, and Filament Elongation[W

    PubMed Central

    Cecchetti, Valentina; Altamura, Maria Maddalena; Falasca, Giuseppina; Costantino, Paolo; Cardarelli, Maura

    2008-01-01

    We provide evidence on the localization, synthesis, transport, and effects of auxin on the processes occurring late in Arabidopsis thaliana stamen development: anther dehiscence, pollen maturation, and preanthesis filament elongation. Expression of auxin-sensitive reporter constructs suggests that auxin effects begin in anthers between the end of meiosis and the bilocular stage in the somatic tissues involved in the first step of dehiscence as well as in the microspores and in the junction region between anther and filament. In situ hybridizations of the auxin biosynthetic genes YUC2 and YUC6 suggest that auxin is synthesized in anthers. In agreement with the timing of auxin effects, the TIR1, AFB1, AFB2, and AFB3 auxin receptor-encoding genes are transcribed in anthers only during late stages of development starting at the end of meiosis. We found that in tir1 afb triple and quadruple mutants, anther dehiscence and pollen maturation occur earlier than in the wild type, causing the release of mature pollen grains before the completion of filament elongation. We also assessed the contribution of auxin transport to late stamen developmental processes. Our results suggest that auxin synthesized in anthers plays a major role in coordinating anther dehiscence and pollen maturation, while auxin transport contributes to the independent regulation of preanthesis filament elongation. PMID:18628351

  15. Tropomodulin isoforms regulate thin filament pointed-end capping and skeletal muscle physiology

    PubMed Central

    Gokhin, David S.; Lewis, Raymond A.; McKeown, Caroline R.; Nowak, Roberta B.; Kim, Nancy E.; Littlefield, Ryan S.; Lieber, Richard L.

    2010-01-01

    During myofibril assembly, thin filament lengths are precisely specified to optimize skeletal muscle function. Tropomodulins (Tmods) are capping proteins that specify thin filament lengths by controlling actin dynamics at pointed ends. In this study, we use a genetic targeting approach to explore the effects of deleting Tmod1 from skeletal muscle. Myofibril assembly, skeletal muscle structure, and thin filament lengths are normal in the absence of Tmod1. Tmod4 localizes to thin filament pointed ends in Tmod1-null embryonic muscle, whereas both Tmod3 and -4 localize to pointed ends in Tmod1-null adult muscle. Substitution by Tmod3 and -4 occurs despite their weaker interactions with striated muscle tropomyosins. However, the absence of Tmod1 results in depressed isometric stress production during muscle contraction, systemic locomotor deficits, and a shift to a faster fiber type distribution. Thus, Tmod3 and -4 compensate for the absence of Tmod1 structurally but not functionally. We conclude that Tmod1 is a novel regulator of skeletal muscle physiology. PMID:20368620

  16. Pathway of actin filament branch formation by Arp2/3 complex revealed by single-molecule imaging

    PubMed Central

    Smith, Benjamin A.; Daugherty-Clarke, Karen; Goode, Bruce L.; Gelles, Jeff

    2013-01-01

    Actin filament nucleation by actin-related protein (Arp) 2/3 complex is a critical process in cell motility and endocytosis, yet key aspects of its mechanism are unknown due to a lack of real-time observations of Arp2/3 complex through the nucleation process. Triggered by the verprolin homology, central, and acidic (VCA) region of proteins in the Wiskott-Aldrich syndrome protein (WASp) family, Arp2/3 complex produces new (daughter) filaments as branches from the sides of preexisting (mother) filaments. We visualized individual fluorescently labeled Arp2/3 complexes dynamically interacting with and producing branches on growing actin filaments in vitro. Branch formation was strikingly inefficient, even in the presence of VCA: only ∼1% of filament-bound Arp2/3 complexes yielded a daughter filament. VCA acted at multiple steps, increasing both the association rate of Arp2/3 complexes with mother filament and the fraction of filament-bound complexes that nucleated a daughter. The results lead to a quantitative kinetic mechanism for branched actin assembly, revealing the steps that can be stimulated by additional cellular factors. PMID:23292935

  17. Physicochemical regulation of biofilm formation

    PubMed Central

    Renner, Lars D.; Weibel, Douglas B.

    2011-01-01

    This article reviews the physical and chemical constraints of environments on biofilm formation. We provide a perspective on how materials science and engineering can address fundamental questions and unmet technological challenges in this area of microbiology, such as biofilm prevention. Specifically, we discuss three factors that impact the development and organization of bacterial communities. (1) Physical properties of surfaces regulate cell attachment and physiology and affect early stages of biofilm formation. (2) Chemical properties influence the adhesion of cells to surfaces and their development into biofilms and communities. (3) Chemical communication between cells attenuates growth and influences the organization of communities. Mechanisms of spatial and temporal confinement control the dimensions of communities and the diffusion path length for chemical communication between biofilms, which, in turn, influences biofilm phenotypes. Armed with a detailed understanding of biofilm formation, researchers are applying the tools and techniques of materials science and engineering to revolutionize the study and control of bacterial communities growing at interfaces. PMID:22125358

  18. Chaperonin filaments: The archael cytoskeleton

    SciTech Connect

    Trent, J.D.; Kagawa, H.K.; Yaoi, Takuro; Olle, E.; Zaluzec, N.J.

    1997-08-01

    Chaperonins are multi-subunit double-ring complexed composed of 60-kDa proteins that are believed to mediate protein folding in vivo. The chaperonins in the hyperthermophilic archaeon Sulfolobus shibatae are composed of the organism`s two most abundant proteins, which represent 4% of its total protein and have an intracellular concentration of {ge} 3.0 mg/ml. At concentrations of 1.0 mg/ml, purified chaperonin proteins aggregate to form ordered filaments. Filament formation, which requires Mg{sup ++} and nucleotide binding (not hydrolysis), occurs at physiological temperatures under conditions suggesting filaments may exist in vivo. If the estimated 4,600 chaperonins per cell, formed filaments in vivo, they could create a matrix of filaments that would span the diameter of an average S. shibatae cell 100 times. Direct observations of unfixed, minimally treated cells by intermediate voltage electron microscopy (300 kV) revealed an intracellular network of filaments that resembles chaperonin filaments produced in vitro. The hypothesis that the intracellular network contains chaperonins is supported by immunogold analyses. The authors propose that chaperonin activity may be regulated in vivo by filament formation and that chaperonin filaments may serve a cytoskeleton-like function in archaea and perhaps in other prokaryotes.

  19. Laser-filamentation-induced water condensation and snow formation in a cloud chamber filled with different ambient gases.

    PubMed

    Liu, Yonghong; Sun, Haiyi; Liu, Jiansheng; Liang, Hong; Ju, Jingjing; Wang, Tiejun; Tian, Ye; Wang, Cheng; Liu, Yi; Chin, See Leang; Li, Ruxin

    2016-04-04

    We investigated femtosecond laser-filamentation-induced airflow, water condensation and snow formation in a cloud chamber filled respectively with air, argon and helium. The mass of snow induced by laser filaments was found being the maximum when the chamber was filled with argon, followed by air and being the minimum with helium. We also discussed the mechanisms of water condensation in different gases. The results show that filaments with higher laser absorption efficiency, which result in higher plasma density, are beneficial for triggering intense airflow and thus more water condensation and precipitation.

  20. The role of Bgl2p in the transition to filamentous cells during biofilm formation by Candida albicans.

    PubMed

    Chen, Xinyue; Zhang, Ruoyu; Takada, Ayako; Iwatani, Shun; Oka, Chiemi; Kitamoto, Toshitaka; Kajiwara, Susumu

    2017-02-01

    The fungal pathogen Candida albicans undergoes a transition from yeast cells to filamentous cells that is related to its pathogenicity. The complex multicellular processes involved in biofilm formation by this fungus also include this transition. In this work, we investigated the morphological role of the Bgl2 protein (Bgl2p) in the transition to filamentous cells during biofilm formation by C. albicans. Bgl2p has been identified as a β-1, 3-glucosyltransferase, and transcription of the CaBGL2 gene is upregulated during biofilm formation. We used scanning electron microscopy to observe the microstructure of a bgl2 null mutant during biofilm formation and found a delay in the transition to filamentous cells in the premature phase (24 hours) of biofilm formation. Deletion of the CaBGL2 gene led to a decrease in the expression of CPH2 and TEC1, which encode transcription factors required for the transition to the filamentous form. These findings indicate that Bgl2p plays a role in the transition to filamentous cells during biofilm formation by C. albicans. © 2016 Blackwell Verlag GmbH.

  1. Light regulation on growth, development, and secondary metabolism of marine-derived filamentous fungi.

    PubMed

    Cai, Menghao; Fang, Zhe; Niu, Chuanpeng; Zhou, Xiangshan; Zhang, Yuanxing

    2013-11-01

    Effects of different light conditions on development, growth, and secondary metabolism of three marine-derived filamentous fungi were investigated. Darkness irritated sexual development of Aspergillus glaucus HB1-19, while white, red, and blue lights improved its asexual behavior. The red and blue lights improved asexual stroma formation of Xylaria sp. (no. 2508), but the darkness and white light inhibited it. Differently, development of Halorosellinia sp. (no. 1403) turned out to be insensitive to any tested light irradiation. Upon the experimental data, no regularity was observed linking development with secondary metabolism. However, fungal growth showed inversely correlation with productions of major bioactive compounds (aspergiolide A, 1403C, and xyloketal B) from various strains. The results indicated that aspergiolide A biosynthesis favored blue light illumination, while 1403C and xyloketal B preferred red light irradiation. With the favorite light sensing conditions, productions of aspergiolide A, 1403C, and xyloketal B were enhanced by 32.9, 21.9, and 30.8 % compared with those in the dark, respectively. The phylogenetic analysis comparing the light-responding proteins of A. glaucus HB 1-19 with those in other systems indicated that A. glaucus HB 1-19 was closely related to Aspergillus spp. especially A. nidulans in spite of its role of marine-derived fungus. It indicated that marine fungi might conserve its light response system when adapting the marine environment. This work also offers useful information for process optimization involving light regulation on growth and metabolism for drug candidate production from light-sensitive marine fungi.

  2. Cdc42p-Interacting Protein Bem4p Regulates the Filamentous-Growth Mitogen-Activated Protein Kinase Pathway

    PubMed Central

    Pitoniak, Andrew; Chavel, Colin A.; Chow, Jacky; Smith, Jeremy; Camara, Diawoye; Karunanithi, Sheelarani; Li, Boyang; Wolfe, Kennith H.

    2014-01-01

    The ubiquitous Rho (Ras homology) GTPase Cdc42p can function in different settings to regulate cell polarity and cellular signaling. How Cdc42p and other proteins are directed to function in a particular context remains unclear. We show that the Cdc42p-interacting protein Bem4p regulates the mitogen-activated protein kinase (MAPK) pathway that controls filamentous growth in Saccharomyces cerevisiae. Bem4p controlled the filamentous-growth pathway but not other MAPK pathways (mating or high-osmolarity glycerol response [HOG]) that also require Cdc42p and other shared components. Bem4p associated with the plasma membrane (PM) protein, Sho1p, to regulate MAPK activity and cell polarization under nutrient-limiting conditions that favor filamentous growth. Bem4p also interacted with the major activator of Cdc42p, the guanine nucleotide exchange factor (GEF) Cdc24p, which we show also regulates the filamentous-growth pathway. Bem4p interacted with the pleckstrin homology (PH) domain of Cdc24p, which functions in an autoinhibitory capacity, and was required, along with other pathway regulators, to maintain Cdc24p at polarized sites during filamentous growth. Bem4p also interacted with the MAPK kinase kinase (MAPKKK) Ste11p. Thus, Bem4p is a new regulator of the filamentous-growth MAPK pathway and binds to general proteins, like Cdc42p and Ste11p, to promote a pathway-specific response. PMID:25384973

  3. Guanyl Nucleotide Exchange Factor Sql2 and Ras2 Regulate Filamentous Growth in Ustilago maydis

    PubMed Central

    Müller, Philip; Katzenberger, Jörg D.; Loubradou , Gabriel; Kahmann, Regine

    2003-01-01

    The cyclic AMP (cAMP)-signaling pathway regulates cell morphology and plays a crucial role during pathogenic development of the plant-pathogenic fungus Ustilago maydis. Strains lacking components of this signaling pathway, such as the Gα-subunit Gpa3 or the adenylyl cyclase Uac1, are nonpathogenic and grow filamentously. On the other hand, strains exhibiting an activated cAMP pathway due to a dominant-active allele of gpa3 display a glossy colony phenotype and are unable to proliferate in plant tumors. Here we present the identification of sql2 as a suppressor of the glossy colony phenotype of a gpa3Q206L strain. sql2 encodes a protein with similarity to CDC25-like guanine nucleotide exchange factors, which are known to act on Ras proteins. Overexpression of sql2 leads to filamentous growth that cannot be suppressed by exogenous cAMP, suggesting that Sql2 does not act upstream of Uac1. To gain more insight in signaling processes regulated by Sql2, we isolated two genes encoding Ras proteins. Expression of dominant active alleles of ras1 and ras2 showed that Ras2 induces filamentous growth while Ras1 does not affect cell morphology but elevates pheromone gene expression. These results indicate that Ras1 and Ras2 fulfill different functions in U. maydis. Moreover, observed similarities between the filaments induced by sql2 and ras2 suggest that Sql2 is an activator of Ras2. Interestingly, sql2 deletion mutants are affected in pathogenic development but not in mating, indicating a specific function of sql2 during pathogenesis. PMID:12796306

  4. Capric Acid Secreted by S. boulardii Inhibits C. albicans Filamentous Growth, Adhesion and Biofilm Formation

    PubMed Central

    Murzyn, Anna; Krasowska, Anna; Stefanowicz, Piotr; Dziadkowiec, Dorota; Łukaszewicz, Marcin

    2010-01-01

    Candidiasis are life-threatening systemic fungal diseases, especially of gastro intestinal track, skin and mucous membranes lining various body cavities like the nostrils, the mouth, the lips, the eyelids, the ears or the genital area. Due to increasing resistance of candidiasis to existing drugs, it is very important to look for new strategies helping the treatment of such fungal diseases. One promising strategy is the use of the probiotic microorganisms, which when administered in adequate amounts confer a health benefit. Such a probiotic microorganism is yeast Saccharomyces boulardii, a close relative of baker yeast. Saccharomyces boulardii cells and their extract affect the virulence factors of the important human fungal pathogen C. albicans, its hyphae formation, adhesion and biofilm development. Extract prepared from S. boulardii culture filtrate was fractionated and GC-MS analysis showed that the active fraction contained, apart from 2-phenylethanol, caproic, caprylic and capric acid whose presence was confirmed by ESI-MS analysis. Biological activity was tested on C. albicans using extract and pure identified compounds. Our study demonstrated that this probiotic yeast secretes into the medium active compounds reducing candidal virulence factors. The chief compound inhibiting filamentous C. albicans growth comparably to S. boulardii extract was capric acid, which is thus responsible for inhibition of hyphae formation. It also reduced candidal adhesion and biofilm formation, though three times less than the extract, which thus contains other factors suppressing C. albicans adherence. The expression profile of selected genes associated with C. albicans virulence by real-time PCR showed a reduced expression of HWP1, INO1 and CSH1 genes in C. albicans cells treated with capric acid and S. boulardii extract. Hence capric acid secreted by S. boulardii is responsible for inhibition of C. albicans filamentation and partially also adhesion and biofilm formation. PMID

  5. AC electric field controlled non-Newtonian filament thinning and droplet formation on the microscale.

    PubMed

    Huang, Y; Wang, Y L; Wong, T N

    2017-08-22

    Monodispersity and fast generation are innate advantages of microfluidic droplets. Other than the normally adopted simple Newtonian fluids such as a water/oil emulsion system, fluids with complex rheology, namely, non-Newtonian fluids, which are being widely adopted in industries and bioengineering, have gained increasing research interest on the microscale. However, challenges occur in controlling the dynamic behavior due to their complex properties. In this sense, the AC electric field with merits of fast response and easiness in fulfilling "Lab on a chip" has attracted our attention. We design and fabricate flow-focusing microchannels with non-contact types of electrodes for the investigation. We firstly compare the formation of a non-Newtonian droplet with that of a Newtonian one under an AC electric field and discover that viscoelasticity contributes to the discrepancies significantly. Then we explore the effect of AC electric fields on the filament thinning and droplet formation dynamics of one non-Newtonian fluid which has a similar rheological behavior to bio samples, such as DNA or blood samples. We investigate the dynamics of the thinning process of the non-Newtonian filament under the influence of an AC electric field and implement a systematic exploration of the non-Newtonian droplet generation influenced by parameters such as the flow conditions (flow rate Q, capillary number Ca), fluid property (Weissenberg number Wi), applied voltage (U) and frequency (f) of the AC electric field. We present the dependencies of the flow condition and electric field on the non-Newtonian droplet formation dynamics, and conclude with an operating diagram, taking into consideration all the above-mentioned parameters. Results show that the electric field plays a critical role in controlling the thinning process of the filament and the size of the generated droplet. Furthermore, for the first time, we quantitatively measure the flow field of the non-Newtonian droplet

  6. Magnetohydrodynamical simulation of the formation of clumps and filaments in quiescent diffuse medium by thermal instability

    NASA Astrophysics Data System (ADS)

    Wareing, C. J.; Pittard, J. M.; Falle, S. A. E. G.; Van Loo, S.

    2016-06-01

    We have used the adaptive mesh refinement hydrodynamic code, MG, to perform idealized 3D magnetohydrodynamical simulations of the formation of clumpy and filamentary structure in a thermally unstable medium without turbulence. A stationary thermally unstable spherical diffuse atomic cloud with uniform density in pressure equilibrium with low density surroundings was seeded with random density variations and allowed to evolve. A range of magnetic field strengths threading the cloud have been explored, from β = 0.1 to 1.0 to the zero magnetic field case (β = ∞), where β is the ratio of thermal pressure to magnetic pressure. Once the density inhomogeneities had developed to the point where gravity started to become important, self-gravity was introduced to the simulation. With no magnetic field, clouds and clumps form within the cloud with aspect ratios of around unity, whereas in the presence of a relatively strong field (β = 0.1) these become filaments, then evolve into interconnected corrugated sheets that are predominantly perpendicular to the magnetic field. With magnetic and thermal pressure equality (β = 1.0), filaments, clouds and clumps are formed. At any particular instant, the projection of the 3D structure on to a plane parallel to the magnetic field, i.e. a line of sight perpendicular to the magnetic field, resembles the appearance of filamentary molecular clouds. The filament densities, widths, velocity dispersions and temperatures resemble those observed in molecular clouds. In contrast, in the strong field case β = 0.1, projection of the 3D structure along a line of sight parallel to the magnetic field reveals a remarkably uniform structure.

  7. Peripheral Leptin Regulates Bone Formation

    PubMed Central

    Turner, Russell T.; Kalra, Satya P.; Wong, Carmen P.; Philbrick, Kenneth A.; Lindenmaier, Laurence B.; Boghossian, Stephane; Iwaniec, Urszula T.

    2012-01-01

    Substantial evidence does not support the prevailing view that leptin, acting through a hypothalamic relay, decreases bone accrual by inhibiting bone formation. To clarify the mechanisms underlying regulation of bone architecture by leptin, we evaluated bone growth and turnover in wild type (WT) mice, leptin receptor-deficient db/db mice, leptin-deficient ob/ob mice and ob/ob mice treated with leptin. We also performed hypothalamic leptin gene therapy to determine the effect of elevated hypothalamic leptin levels on osteoblasts. Finally, to determine the effects of loss of peripheral leptin signaling on bone formation and energy metabolism, we used bone marrow (BM) from WT or db/db donor mice to reconstitute the hematopoietic and mesenchymal stem cell compartments in lethally irradiated WT recipient mice. Decreases in bone growth, osteoblast-lined bone perimeter and bone formation rate were observed in ob/ob mice and greatly increased in ob/ob mice following subcutaneous administration of leptin. Similarly, hypothalamic leptin gene therapy increased osteoblast-lined bone perimeter in ob/ob mice. In spite of normal osteoclast-lined bone perimeter, db/db mice exhibited a mild but generalized osteopetrotic-like (calcified cartilage encased by bone) skeletal phenotype and greatly reduced serum markers of bone turnover. Tracking studies and histology revealed quantitative replacement of BM cells following BM transplantation. WT mice engrafted with db/db BM did not differ in energy homeostasis from untreated WT mice or WT mice engrafted with WT BM. Bone formation in WT mice engrafted with WT BM did not differ from WT mice, whereas bone formation in WT mice engrafted with db/db cells did not differ from the low rates observed in untreated db/db mice. In summary, our results indicate that leptin, acting primarily through peripheral pathways, increases osteoblast number and activity. PMID:22887758

  8. Ambipolar diffusion regulated collapse of filaments threaded by perpendicular magnetic fields

    NASA Astrophysics Data System (ADS)

    Burge, C. A.; Van Loo, S.; Falle, S. A. E. G.; Hartquist, T. W.

    2016-11-01

    the collapse is governed by magnetically-regulated ambipolar diffusion. The gas collapses at velocities much lower than the sound speed. For X ≲ 10-8, the gas is weakly coupled to the magnetic field and the magnetic support is removed by gravitationally-dominated ambipolar diffusion. Here, neutrals and ions only collide sporadically, that is the ambipolar diffusion length scale is larger than the Jeans length, and the gas can attain high collapse velocities. When decaying turbulence is included, additional support is provided to the filament. This slows down the collapse of the filament even in the absence of a magnetic field. When a magnetic field is present, the collapse rate increases by a ratio smaller than for the non-magnetic case. This is because of a speed-up of the ambipolar diffusion due to larger magnetic field gradients generated by the turbulence and because the ambipolar diffusion aids the dissipation of turbulence below the ambipolar diffusion length scale. The highest increase in the rate is observed for the lowest ionisation coefficient and the highest turbulent intensity.

  9. The GRF10 homeobox gene regulates filamentous growth in the human fungal pathogen Candida albicans.

    PubMed

    Ghosh, Anup K; Wangsanut, Tanaporn; Fonzi, William A; Rolfes, Ronda J

    2015-12-01

    Candida albicans is the most common human fungal pathogen and can cause life-threatening infections. Filamentous growth is critical in the pathogenicity of C. albicans, as the transition from yeast to hyphal forms is linked to virulence and is also a pivotal process in fungal biofilm development. Homeodomain-containing transcription factors have been linked to developmental processes in fungi and other eukaryotes. We report here on GRF10, a homeobox transcription factor-encoding gene that plays a role in C. albicans filamentation. Deletion of the GRF10 gene, in both C. albicans SN152 and BWP17 strain backgrounds, results in mutants with strongly decreased hyphal growth. The mutants are defective in chlamydospore and biofilm formation, as well as showing dramatically attenuated virulence in a mouse infection model. Expression of the GRF10 gene is highly induced during stationary phase and filamentation. In summary, our study emphasizes a new role for the homeodomain-containing transcription factor in morphogenesis and pathogenicity of C. albicans.

  10. Mechanical output of myosin II motors is regulated by myosin filament size and actin network mechanics

    NASA Astrophysics Data System (ADS)

    Stam, Samantha; Alberts, Jonathan; Gardel, Margaret; Munro, Edwin

    2013-03-01

    The interactions of bipolar myosin II filaments with actin arrays are a predominate means of generating forces in numerous physiological processes including muscle contraction and cell migration. However, how the spatiotemporal regulation of these forces depends on motor mechanochemistry, bipolar filament size, and local actin mechanics is unknown. Here, we simulate myosin II motors with an agent-based model in which the motors have been benchmarked against experimental measurements. Force generation occurs in two distinct regimes characterized either by stable tension maintenance or by stochastic buildup and release; transitions between these regimes occur by changes to duty ratio and myosin filament size. The time required for building force to stall scales inversely with the stiffness of a network and the actin gliding speed of a motor. Finally, myosin motors are predicted to contract a network toward stiffer regions, which is consistent with experimental observations. Our representation of myosin motors can be used to understand how their mechanical and biochemical properties influence their observed behavior in a variety of in vitro and in vivo contexts.

  11. Actin-crosslinking protein regulation of filament movement in motility assays: a theoretical model.

    PubMed Central

    Janson, L W; Taylor, D L

    1994-01-01

    The interaction of single actin filaments on a myosin-coated coverslip has been modeled by several authors. One model adds a component of "frictional drag" by myosin heads that oppose movement of the actin filaments. We have extended this concept by including the resistive drag from actin crosslinking proteins to understand better the relationship among crosslinking number, actin-myosin force generation, and motility. The validity of this model is supported by agreement with the experimental results from a previous study in which crosslinking proteins were added with myosin molecules under otherwise standard motility assay conditions. The theoretical relationship provides a means to determine many physical parameters that characterize the interaction between a single actin filament and a single actin-crosslinking molecule (various types). In particular, the force constant of a single filamin molecule is calculated as 1.105 pN, approximately 3 times less than a driving myosin head (3.4 pN). Knowledge of this parameter and others derived from this model allows a better understanding of the interaction between myosin and the actin/actin-binding protein cytoskeleton and the role of actin-binding proteins in the regulation and modulation of motility. PMID:7811954

  12. Low-power resistive random access memory by confining the formation of conducting filaments

    SciTech Connect

    Huang, Yi-Jen; Lee, Si-Chen; Shen, Tzu-Hsien; Lee, Lan-Hsuan; Wen, Cheng-Yen

    2016-06-15

    Owing to their small physical size and low power consumption, resistive random access memory (RRAM) devices are potential for future memory and logic applications in microelectronics. In this study, a new resistive switching material structure, TiO{sub x}/silver nanoparticles/TiO{sub x}/AlTiO{sub x}, fabricated between the fluorine-doped tin oxide bottom electrode and the indium tin oxide top electrode is demonstrated. The device exhibits excellent memory performances, such as low operation voltage (<±1 V), low operation power, small variation in resistance, reliable data retention, and a large memory window. The current-voltage measurement shows that the conducting mechanism in the device at the high resistance state is via electron hopping between oxygen vacancies in the resistive switching material. When the device is switched to the low resistance state, conducting filaments are formed in the resistive switching material as a result of accumulation of oxygen vacancies. The bottom AlTiO{sub x} layer in the device structure limits the formation of conducting filaments; therefore, the current and power consumption of device operation are significantly reduced.

  13. Metallic filament formation by aligned oxygen vacancies in ZnO-based resistive switches

    SciTech Connect

    Gu, Tingkun

    2014-05-28

    The electronic structure of ZnO with defects of oxygen vacancies were investigated by using first-principles methods. Some structure models were constructed in order to investigate the effects of the distribution of oxygen vacancies on the electronic properties of ZnO. By analyzing the calculated results, we found that only the aligned oxygen vacancies can form the conducting channel in ZnO, and the transformation of the oxygen vacancy from charged state to neutral state is consistent with the energetics rule of the forming aligned oxygen vacancies. As for the heterojunction of Pt/ZnO/Pt, the oxygen vacancies near the interface of Pt/ZnO depress the local Schottky barrier effectively, and the aligned oxygen vacancies in ZnO form a conducting filament connecting two Pt electrodes. The metallic filament formation in Pt/ZnO/Pt resistive switching cells should be closely related to the carrier injection from Pt electrode into ZnO and the arrangement of oxygen vacancies in ZnO slab.

  14. Polar pattern formation in driven filament systems requires non-binary particle collisions

    NASA Astrophysics Data System (ADS)

    Suzuki, Ryo; Weber, Christoph A.; Frey, Erwin; Bausch, Andreas R.

    2015-10-01

    From the self-organization of the cytoskeleton to the synchronous motion of bird flocks, living matter has the extraordinary ability to behave in a concerted manner. The Boltzmann equation for self-propelled particles is frequently used in silico to link a system’s meso- or macroscopic behaviour to the microscopic dynamics of its constituents. But so far such studies have relied on an assumption of simplified binary collisions owing to a lack of experimental data suggesting otherwise. We report here experimentally determined binary-collision statistics by studying a recently introduced molecular system, the high-density actomyosin motility assay. We demonstrate that the alignment induced by binary collisions is too weak to account for the observed ordering transition. The transition density for polar pattern formation decreases quadratically with filament length, indicating that multi-filament collisions drive the observed ordering phenomenon and that a gas-like picture cannot explain the transition of the system to polar order. Our findings demonstrate that the unique properties of biological active-matter systems require a description that goes well beyond that developed in the framework of kinetic theories.

  15. Prostaglandins temporally regulate cytoplasmic actin bundle formation during Drosophila oogenesis.

    PubMed

    Spracklen, Andrew J; Kelpsch, Daniel J; Chen, Xiang; Spracklen, Cassandra N; Tootle, Tina L

    2014-02-01

    Prostaglandins (PGs)--lipid signals produced downstream of cyclooxygenase (COX) enzymes--regulate actin dynamics in cell culture and platelets, but their roles during development are largely unknown. Here we define a new role for Pxt, the Drosophila COX-like enzyme, in regulating the actin cytoskeleton--temporal restriction of actin remodeling during oogenesis. PGs are required for actin filament bundle formation during stage 10B (S10B). In addition, loss of Pxt results in extensive early actin remodeling, including actin filaments and aggregates, within the posterior nurse cells of S9 follicles; wild-type follicles exhibit similar structures at a low frequency. Hu li tai shao (Hts-RC) and Villin (Quail), an actin bundler, localize to all early actin structures, whereas Enabled (Ena), an actin elongation factor, preferentially localizes to those in pxt mutants. Reduced Ena levels strongly suppress early actin remodeling in pxt mutants. Furthermore, loss of Pxt results in reduced Ena localization to the sites of bundle formation during S10B. Together these data lead to a model in which PGs temporally regulate actin remodeling during Drosophila oogenesis by controlling Ena localization/activity, such that in S9, PG signaling inhibits, whereas at S10B, it promotes Ena-dependent actin remodeling.

  16. Large-Scale Patterns of Filament Channels and Filaments

    NASA Astrophysics Data System (ADS)

    Mackay, Duncan

    2016-07-01

    In this review the properties and large-scale patterns of filament channels and filaments will be considered. Initially, the global formation locations of filament channels and filaments are discussed, along with their hemispheric pattern. Next, observations of the formation of filament channels and filaments are described where two opposing views are considered. Finally, the wide range of models that have been constructed to consider the formation of filament channels and filaments over long time-scales are described, along with the origin of the hemispheric pattern of filaments.

  17. RhoA Signaling Is Required for Respiratory Syncytial Virus-Induced Syncytium Formation and Filamentous Virion Morphology

    PubMed Central

    Gower, Tara L.; Pastey, Manoj K.; Peeples, Mark E.; Collins, Peter L.; McCurdy, Lewis H.; Hart, Timothy K.; Guth, Alex; Johnson, Teresa R.; Graham, Barney S.

    2005-01-01

    Respiratory syncytial virus (RSV) is an important human pathogen that can cause severe and life-threatening respiratory infections in infants, the elderly, and immunocompromised adults. RSV infection of HEp-2 cells induces the activation of RhoA, a small GTPase. We therefore asked whether RhoA signaling is important for RSV replication or syncytium formation. The treatment of HEp-2 cells with Clostridium botulinum C3, an enzyme that ADP-ribosylates and specifically inactivates RhoA, inhibited RSV-induced syncytium formation and cell-to-cell fusion, although similar levels of PFU were released into the medium and viral protein expression levels were equivalent. Treatment with another inhibitor of RhoA signaling, the Rho kinase inhibitor Y-27632, yielded similar results. Scanning electron microscopy of C3-treated infected cells showed reduced numbers of single blunted filaments, in contrast to the large clumps of long filaments in untreated infected cells. These data suggest that RhoA signaling is associated with filamentous virus morphology, cell-to-cell fusion, and syncytium formation but is dispensable for the efficient infection and production of infectious virus in vitro. Next, we developed a semiquantitative method to measure spherical and filamentous virus particles by using sucrose gradient velocity sedimentation. Fluorescence and transmission electron microscopy confirmed the separation of spherical and filamentous forms of infectious virus into two identifiable peaks. The C3 treatment of RSV-infected cells resulted in a shift to relatively more spherical virions than those from untreated cells. These data suggest that viral filamentous protuberances characteristic of RSV infection are associated with RhoA signaling, are important for filamentous virion morphology, and may play a role in initiating cell-to-cell fusion. PMID:15827147

  18. The Amphipathic Helix of Influenza A Virus M2 Protein Is Required for Filamentous Bud Formation and Scission of Filamentous and Spherical Particles

    PubMed Central

    Roberts, Kari L.; Leser, George P.; Ma, Chunlong

    2013-01-01

    Influenza virus assembles and buds at the infected-cell plasma membrane. This involves extrusion of the plasma membrane followed by scission of the bud, resulting in severing the nascent virion from its former host. The influenza virus M2 ion channel protein contains in its cytoplasmic tail a membrane-proximal amphipathic helix that facilitates the scission process and is also required for filamentous particle formation. Mutation of five conserved hydrophobic residues to alanines within the amphipathic helix (M2 five-point mutant, or 5PM) reduced scission and also filament formation, whereas single mutations had no apparent phenotype. Here, we show that any two of these five residues mutated together to alanines result in virus debilitated for growth and filament formation in a manner similar to 5PM. Growth kinetics of the M2 mutants are approximately 2 logs lower than the wild-type level, and plaque diameter was significantly reduced. When the 5PM and a representative double mutant (I51A-Y52A) were introduced into A/WSN/33 M2, a strain that produces spherical particles, similar debilitation in viral growth occurred. Electron microscopy showed that with the 5PM and the I51A-Y52A A/Udorn/72 and WSN viruses, scission failed, and emerging virus particles exhibited a “beads-on-a-string” morphology. The major spike glycoprotein hemagglutinin is localized within lipid rafts in virus-infected cells, whereas M2 is associated at the periphery of rafts. Mutant M2s were more widely dispersed, and their abundance at the raft periphery was reduced, suggesting that the M2 amphipathic helix is required for proper localization in the host membrane and that this has implications for budding and scission. PMID:23843641

  19. The amphipathic helix of influenza A virus M2 protein is required for filamentous bud formation and scission of filamentous and spherical particles.

    PubMed

    Roberts, Kari L; Leser, George P; Ma, Chunlong; Lamb, Robert A

    2013-09-01

    Influenza virus assembles and buds at the infected-cell plasma membrane. This involves extrusion of the plasma membrane followed by scission of the bud, resulting in severing the nascent virion from its former host. The influenza virus M2 ion channel protein contains in its cytoplasmic tail a membrane-proximal amphipathic helix that facilitates the scission process and is also required for filamentous particle formation. Mutation of five conserved hydrophobic residues to alanines within the amphipathic helix (M2 five-point mutant, or 5PM) reduced scission and also filament formation, whereas single mutations had no apparent phenotype. Here, we show that any two of these five residues mutated together to alanines result in virus debilitated for growth and filament formation in a manner similar to 5PM. Growth kinetics of the M2 mutants are approximately 2 logs lower than the wild-type level, and plaque diameter was significantly reduced. When the 5PM and a representative double mutant (I51A-Y52A) were introduced into A/WSN/33 M2, a strain that produces spherical particles, similar debilitation in viral growth occurred. Electron microscopy showed that with the 5PM and the I51A-Y52A A/Udorn/72 and WSN viruses, scission failed, and emerging virus particles exhibited a "beads-on-a-string" morphology. The major spike glycoprotein hemagglutinin is localized within lipid rafts in virus-infected cells, whereas M2 is associated at the periphery of rafts. Mutant M2s were more widely dispersed, and their abundance at the raft periphery was reduced, suggesting that the M2 amphipathic helix is required for proper localization in the host membrane and that this has implications for budding and scission.

  20. Mammalian homolog of the yeast cyclase associated protein, CAP/Srv2p, regulates actin filament assembly.

    PubMed

    Freeman, N L; Field, J

    2000-02-01

    Control of cell shape and motility requires rearrangements of the actin cytoskeleton. One cytoskeletal protein that may regulate actin dynamics is CAP (cyclase associated protein; CAP/Srv2p; ASP-56). CAP was first isolated from yeast as an adenylyl cyclase associated protein required for RAS regulation of cAMP signaling. In addition, CAP also regulates the actin cytoskeleton primarily through an actin monomer binding activity. CAP homologs are found in many eukaryotes, including mammals where they also bind actin, but little is known about their biological function. We, therefore, designed experiments to address CAP1 regulation of the actin cytoskeleton. CAP1 localized to membrane ruffles and actin stress fibers in fixed cells of various types. To address localization in living cells, we constructed GFP-CAP1 fusion proteins and found that fusion proteins lacking the actin-binding region localized like the wild type protein. We also performed microinjection studies with affinity-purified anti-CAP1 antibodies in Swiss 3T3 fibroblasts and found that the antibodies attenuated serum stimulation of stress fibers. Finally, CAP1 purified from platelets through a monoclonal antibody affinity purification step stimulated the formation of stress fiber-like filaments when it was microinjected into serum-starved Swiss 3T3 cells. Taken together, these data suggest that CAP1 promotes assembly of the actin cytoskeleton.

  1. Bundle formation of smooth muscle desmin intermediate filaments by calponin and its binding site on the desmin molecule.

    PubMed

    Fujii, T; Takagi, H; Arimoto, M; Ootani, H; Ueeda, T

    2000-03-01

    Smooth muscle basic calponin, a major actin-, tropomyosin-, and calmodulin-binding protein, has been examined for its ability to interact with desmin intermediate filaments from smooth muscle cells using sedimentation analysis, turbidity changes, chemical cross-linking, matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI TOF/MS), and electron microscopic observations. Calponin interacted with desmin intermediate filaments in a concentration-dependent manner in vitro. The binding of calponin to desmin produced dense aggregates at 30 degrees C. The dense aggregates were observed by electron microscopy to be composed of large anisotropic bundles of desmin filaments, indicating that calponin forms bundles of desmin filaments. The addition of calmodulin or S100 to the mixture of calponin and desmin caused the removal of calponin from the desmin filaments and inhibited bundle formation in the presence of Ca(2+), but not in the presence of EGTA. Calponin-related proteins including G-actin, tropomyosin, and SM22, had little effect on the binding of calponin to desmin filaments, whereas tubulin weakly inhibited the binding. Desmin had little influence on the calponin-actin and calponin-tubulin interactions using the zero-length cross-linker, EDC. Domain mapping with chymotryptic digestion showed that the binding site of calponin resides within the central a-helical rod domain of the desmin molecule. The chemical cross-linked products of calponin and synthetic peptides (TQ27, TNEKVELQELNDRFANYIEKVRFLEQQ; EE24, EEELRELRRQVDALTGQRARVEVE) derived from the rod domain were detected by MALDI TOF/MS. Furthermore, the calponin-desmin interaction was significantly inhibited by the addition of EE24, but only slightly by TQ27. These results suggest that calponin may act as a cross-linking protein between desmin filaments as well as among intermediate filaments, microfilaments and microtubules in smooth muscle cells.

  2. Spatiotemporal distribution of different extracellular polymeric substances and filamentation mediate Xylella fastidiosa adhesion and biofilm formation

    PubMed Central

    Janissen, Richard; Murillo, Duber M.; Niza, Barbara; Sahoo, Prasana K.; Nobrega, Marcelo M.; Cesar, Carlos L.; Temperini, Marcia L. A.; Carvalho, Hernandes F.; de Souza, Alessandra A.; Cotta, Monica A.

    2015-01-01

    Microorganism pathogenicity strongly relies on the generation of multicellular assemblies, called biofilms. Understanding their organization can unveil vulnerabilities leading to potential treatments; spatially and temporally-resolved comprehensive experimental characterization can provide new details of biofilm formation, and possibly new targets for disease control. Here, biofilm formation of economically important phytopathogen Xylella fastidiosa was analyzed at single-cell resolution using nanometer-resolution spectro-microscopy techniques, addressing the role of different types of extracellular polymeric substances (EPS) at each stage of the entire bacterial life cycle. Single cell adhesion is caused by unspecific electrostatic interactions through proteins at the cell polar region, where EPS accumulation is required for more firmly-attached, irreversibly adhered cells. Subsequently, bacteria form clusters, which are embedded in secreted loosely-bound EPS, and bridged by up to ten-fold elongated cells that form the biofilm framework. During biofilm maturation, soluble EPS forms a filamentous matrix that facilitates cell adhesion and provides mechanical support, while the biofilm keeps anchored by few cells. This floating architecture maximizes nutrient distribution while allowing detachment upon larger shear stresses; it thus complies with biological requirements of the bacteria life cycle. Using new approaches, our findings provide insights regarding different aspects of the adhesion process of X. fastidiosa and biofilm formation. PMID:25891045

  3. Spatiotemporal distribution of different extracellular polymeric substances and filamentation mediate Xylella fastidiosa adhesion and biofilm formation.

    PubMed

    Janissen, Richard; Murillo, Duber M; Niza, Barbara; Sahoo, Prasana K; Nobrega, Marcelo M; Cesar, Carlos L; Temperini, Marcia L A; Carvalho, Hernandes F; de Souza, Alessandra A; Cotta, Monica A

    2015-04-20

    Microorganism pathogenicity strongly relies on the generation of multicellular assemblies, called biofilms. Understanding their organization can unveil vulnerabilities leading to potential treatments; spatially and temporally-resolved comprehensive experimental characterization can provide new details of biofilm formation, and possibly new targets for disease control. Here, biofilm formation of economically important phytopathogen Xylella fastidiosa was analyzed at single-cell resolution using nanometer-resolution spectro-microscopy techniques, addressing the role of different types of extracellular polymeric substances (EPS) at each stage of the entire bacterial life cycle. Single cell adhesion is caused by unspecific electrostatic interactions through proteins at the cell polar region, where EPS accumulation is required for more firmly-attached, irreversibly adhered cells. Subsequently, bacteria form clusters, which are embedded in secreted loosely-bound EPS, and bridged by up to ten-fold elongated cells that form the biofilm framework. During biofilm maturation, soluble EPS forms a filamentous matrix that facilitates cell adhesion and provides mechanical support, while the biofilm keeps anchored by few cells. This floating architecture maximizes nutrient distribution while allowing detachment upon larger shear stresses; it thus complies with biological requirements of the bacteria life cycle. Using new approaches, our findings provide insights regarding different aspects of the adhesion process of X. fastidiosa and biofilm formation.

  4. Large-scale filament formation inhibits the activity of CTP synthetase

    PubMed Central

    Barry, Rachael M; Bitbol, Anne-Florence; Lorestani, Alexander; Charles, Emeric J; Habrian, Chris H; Hansen, Jesse M; Li, Hsin-Jung; Baldwin, Enoch P; Wingreen, Ned S; Kollman, Justin M; Gitai, Zemer

    2014-01-01

    CTP Synthetase (CtpS) is a universally conserved and essential metabolic enzyme. While many enzymes form small oligomers, CtpS forms large-scale filamentous structures of unknown function in prokaryotes and eukaryotes. By simultaneously monitoring CtpS polymerization and enzymatic activity, we show that polymerization inhibits activity, and CtpS's product, CTP, induces assembly. To understand how assembly inhibits activity, we used electron microscopy to define the structure of CtpS polymers. This structure suggests that polymerization sterically hinders a conformational change necessary for CtpS activity. Structure-guided mutagenesis and mathematical modeling further indicate that coupling activity to polymerization promotes cooperative catalytic regulation. This previously uncharacterized regulatory mechanism is important for cellular function since a mutant that disrupts CtpS polymerization disrupts E. coli growth and metabolic regulation without reducing CTP levels. We propose that regulation by large-scale polymerization enables ultrasensitive control of enzymatic activity while storing an enzyme subpopulation in a conformationally restricted form that is readily activatable. DOI: http://dx.doi.org/10.7554/eLife.03638.001 PMID:25030911

  5. Large-scale filament formation inhibits the activity of CTP synthetase.

    PubMed

    Barry, Rachael M; Bitbol, Anne-Florence; Lorestani, Alexander; Charles, Emeric J; Habrian, Chris H; Hansen, Jesse M; Li, Hsin-Jung; Baldwin, Enoch P; Wingreen, Ned S; Kollman, Justin M; Gitai, Zemer

    2014-07-16

    CTP Synthetase (CtpS) is a universally conserved and essential metabolic enzyme. While many enzymes form small oligomers, CtpS forms large-scale filamentous structures of unknown function in prokaryotes and eukaryotes. By simultaneously monitoring CtpS polymerization and enzymatic activity, we show that polymerization inhibits activity, and CtpS's product, CTP, induces assembly. To understand how assembly inhibits activity, we used electron microscopy to define the structure of CtpS polymers. This structure suggests that polymerization sterically hinders a conformational change necessary for CtpS activity. Structure-guided mutagenesis and mathematical modeling further indicate that coupling activity to polymerization promotes cooperative catalytic regulation. This previously uncharacterized regulatory mechanism is important for cellular function since a mutant that disrupts CtpS polymerization disrupts E. coli growth and metabolic regulation without reducing CTP levels. We propose that regulation by large-scale polymerization enables ultrasensitive control of enzymatic activity while storing an enzyme subpopulation in a conformationally restricted form that is readily activatable.

  6. Intermediate Filaments Play a Pivotal Role in Regulating Cell Architecture and Function*

    PubMed Central

    Lowery, Jason; Kuczmarski, Edward R.; Herrmann, Harald; Goldman, Robert D.

    2015-01-01

    Intermediate filaments (IFs) are composed of one or more members of a large family of cytoskeletal proteins, whose expression is cell- and tissue type-specific. Their importance in regulating the physiological properties of cells is becoming widely recognized in functions ranging from cell motility to signal transduction. IF proteins assemble into nanoscale biopolymers with unique strain-hardening properties that are related to their roles in regulating the mechanical integrity of cells. Furthermore, mutations in the genes encoding IF proteins cause a wide range of human diseases. Due to the number of different types of IF proteins, we have limited this short review to cover structure and function topics mainly related to the simpler homopolymeric IF networks composed of vimentin, and specifically for diseases, the related muscle-specific desmin IF networks. PMID:25957409

  7. Gas7b (growth arrest specific protein 7b) regulates neuronal cell morphology by enhancing microtubule and actin filament assembly.

    PubMed

    Gotoh, Aina; Hidaka, Masafumi; Hirose, Keiko; Uchida, Takafumi

    2013-11-29

    Neurons undergo several morphological changes as a part of normal neuron maturation process. Alzheimer disease is associated with increased neuroproliferation and impaired neuronal maturation. In this study, we demonstrated that Gas7b (growth arrest specific protein 7b) expression in a neuronal cell line, Neuro 2A, induces cell maturation by facilitating formation of dendrite-like processes and/or filopodia projections and that Gas7b co-localizes with neurite microtubules. Molecular analysis was performed to evaluate whether Gas7b associates with actin filaments and microtubules, and the data revealed two novel roles of Gas7b in neurite outgrowth: we showed that Gas7b enhances bundling of several microtubule filaments and connects microtubules with actin filaments. These results suggest that Gas7b governs neural cell morphogenesis by enhancing the coordination between actin filaments and microtubules. We conclude that lower neuronal Gas7b levels may impact Alzheimer disease progression.

  8. Formation and Destabilization of Actin Filaments with Tetramethylrhodamine-Modified Actin

    PubMed Central

    Kudryashov, Dmitry S.; Phillips, Martin; Reisler, Emil

    2004-01-01

    Actin labeling at Cys374 with tethramethylrhodamine derivatives (TMR-actin) has been widely used for direct observation of the in vitro filaments growth, branching, and treadmilling, as well as for the in vivo visualization of actin cytoskeleton. The advantage of TMR-actin is that it does not lock actin in filaments (as rhodamine-phalloidin does), possibly allowing for its use in investigating the dynamic assembly behavior of actin polymers. Although it is established that TMR-actin alone is polymerization incompetent, the impact of its copolymerization with unlabeled actin on filament structure and dynamics has not been tested yet. In this study, we show that TMR-actin perturbs the filaments structure when copolymerized with unlabeled actin; the resulting filaments are more fragile and shorter than the control filaments. Due to the increased severing of copolymer filaments, TMR-actin accelerates the polymerization of unlabeled actin in solution also at mole ratios lower than those used in most fluorescence microscopy experiments. The destabilizing and severing effect of TMR-actin is countered by filament stabilizing factors, phalloidin, S1, and tropomyosin. These results point to an analogy between the effects of TMR-actin and severing proteins on F-actin, and imply that TMR-actin may be inappropriate for investigations of actin filaments dynamics. PMID:15298916

  9. Complex regulation of hydrolytic enzyme genes for cellulosic biomass degradation in filamentous fungi.

    PubMed

    Tani, Shuji; Kawaguchi, Takashi; Kobayashi, Tetsuo

    2014-06-01

    Filamentous fungi produce cellulolytic and hemicellulolytic enzymes in response to small inducer molecules liberated from cellulosic biomass. Enzyme production is mainly regulated at the level of transcription. The first transcription factor identified as being involved in cellulosic biomass degradation was XlnR, which mediates D-xylose-triggered induction of xylanolytic and cellulolytic genes in Aspergillus. XlnR has played the leading role for over a decade in studies aimed at clarification of gene regulation related to cellulosic biomass degradation. Very recently, several new transcription factors were identified, namely Clr-1/2 in Neurospora; ManR, McmA, and ClbR in Aspergillus; and BglR in Trichoderma, all of which participate in the regulation of cellulolytic and/or hemicellulolytic enzyme production. Furthermore, as well as the carbon sources available, other factors such as light signaling and anti-sense RNA accumulation have been shown to contribute to this regulation. Here, we review the recent advancements demonstrating that multiple factors coordinately regulate the expression of cellulosic biomass degrading enzyme genes.

  10. Coupling of transcriptional response to oxidative stress and secondary metabolism regulation in filamentous fungi.

    PubMed

    Montibus, Mathilde; Pinson-Gadais, Laëtitia; Richard-Forget, Florence; Barreau, Christian; Ponts, Nadia

    2015-01-01

    To survive sudden and potentially lethal changes in their environment, filamentous fungi must sense and respond to a vast array of stresses, including oxidative stresses. The generation of reactive oxygen species, or ROS, is an inevitable aspect of existence under aerobic conditions. In addition, in the case of fungi with pathogenic lifestyles, ROS are produced by the infected hosts and serve as defense weapons via direct toxicity, as well as effectors in fungal cell death mechanisms. Filamentous fungi have thus developed complex and sophisticated responses to evade oxidative killing. Several steps are determinant in these responses, including the activation of transcriptional regulators involved in the control of the antioxidant machinery. Gathering and integrating the most recent advances in knowledge of oxidative stress responses in fungi are the main objectives of this review. Most of the knowledge coming from two models, the yeast Saccharomyces cerevisiae and fungi of the genus Aspergillus, is summarized. Nonetheless, recent information on various other fungi is delivered when available. Finally, special attention is given on the potential link between the functional interaction between oxidative stress and secondary metabolism that has been suggested in recent reports, including the production of mycotoxins.

  11. Formation of a compound flux rope by the merging of two filament channels, the associated dynamics, and its stability

    SciTech Connect

    Joshi, Navin Chandra; Inoue, Satoshi; Magara, Tetsuya E-mail: njoshi98@gmail.com

    2014-11-01

    We present observations of compound flux rope formation, which occurred on 2014 January 1, via merging of two nearby filament channels, the associated dynamics, and its stability using multiwavelength data. We also discuss the dynamics of cool and hot plasma moving along the newly formed compound flux rope. The merging started after the interaction between the southern leg of the northward filament and the northern leg of the southward filament at ≈01:21 UT and continued until a compound flux rope formed at ≈01:33 UT. During the merging, the cool filament plasma heated up and started to move along both sides of the compound flux rope, i.e., toward the north (≈265 km s{sup –1}) and south (≈118 km s{sup –1}) from the point of merging. After traveling a distance of ≈150 Mm toward the north, the plasma cooled down and started to return back to the south (≈14 km s{sup –1}) after ≈02:00 UT. The observations provide a clear example of compound flux rope formation via merging of two different flux ropes and the occurrence of a flare through tether cutting reconnection. However, the compound flux rope remained stable in the corona and had a confined eruption. The coronal magnetic field decay index measurements revealed that both the filaments and the compound flux rope axis lie within the stability domain (decay index <1.5), which may be the possible cause for their stability. The present study also deals with the relationship between the filament's chirality (sinistral) and the helicity (positive) of the surrounding flux rope.

  12. DNA Double-Strand Breaks Induce the Nuclear Actin Filaments Formation in Cumulus-Enclosed Oocytes but Not in Denuded Oocytes.

    PubMed

    Sun, Ming-Hong; Yang, Mo; Xie, Feng-Yun; Wang, Wei; Zhang, Lili; Shen, Wei; Yin, Shen; Ma, Jun-Yu

    2017-01-01

    As a gamete, oocyte needs to maintain its genomic integrity and passes this haploid genome to the next generation. However, fully-grown mouse oocyte cannot respond to DNA double-strand breaks (DSBs) effectively and it is also unable to repair them before the meiosis resumption. To compensate for this disadvantage and control the DNA repair events, oocyte needs the cooperation with its surrounding cumulus cells. Recently, evidences have shown that nuclear actin filament formation plays roles in cellular DNA DSB repair. To explore whether these nuclear actin filaments are formed in the DNA-damaged oocytes, here, we labeled the filament actins in denuded oocytes (DOs) and cumulus-enclosed oocytes (CEOs). We observed that the nuclear actin filaments were formed only in the DNA-damaged CEOs, but not in DOs. Formation of actin filaments in the nucleus was an event downstream to the DNA damage response. Our data also showed that the removal of cumulus cells led to a reduction in the nuclear actin filaments in oocytes. Knocking down of the Adcy1 gene in cumulus cells did not affect the formation of nuclear actin filaments in oocytes. Notably, we also observed that the nuclear actin filaments in CEOs could be induced by inhibition of gap junctions. From our results, it was confirmed that DNA DSBs induce the nuclear actin filament formation in oocyte and which is controlled by the cumulus cells.

  13. DNA Double-Strand Breaks Induce the Nuclear Actin Filaments Formation in Cumulus-Enclosed Oocytes but Not in Denuded Oocytes

    PubMed Central

    Sun, Ming-Hong; Yang, Mo; Xie, Feng-Yun; Wang, Wei; Zhang, Lili; Shen, Wei; Yin, Shen

    2017-01-01

    As a gamete, oocyte needs to maintain its genomic integrity and passes this haploid genome to the next generation. However, fully-grown mouse oocyte cannot respond to DNA double-strand breaks (DSBs) effectively and it is also unable to repair them before the meiosis resumption. To compensate for this disadvantage and control the DNA repair events, oocyte needs the cooperation with its surrounding cumulus cells. Recently, evidences have shown that nuclear actin filament formation plays roles in cellular DNA DSB repair. To explore whether these nuclear actin filaments are formed in the DNA-damaged oocytes, here, we labeled the filament actins in denuded oocytes (DOs) and cumulus-enclosed oocytes (CEOs). We observed that the nuclear actin filaments were formed only in the DNA-damaged CEOs, but not in DOs. Formation of actin filaments in the nucleus was an event downstream to the DNA damage response. Our data also showed that the removal of cumulus cells led to a reduction in the nuclear actin filaments in oocytes. Knocking down of the Adcy1 gene in cumulus cells did not affect the formation of nuclear actin filaments in oocytes. Notably, we also observed that the nuclear actin filaments in CEOs could be induced by inhibition of gap junctions. From our results, it was confirmed that DNA DSBs induce the nuclear actin filament formation in oocyte and which is controlled by the cumulus cells. PMID:28099474

  14. CASEIN KINASE1-LIKE PROTEIN2 Regulates Actin Filament Stability and Stomatal Closure via Phosphorylation of Actin Depolymerizing Factor

    PubMed Central

    Zhao, Shuangshuang; Jiang, Yuxiang; Zhao, Yang; Huang, Shanjin; Yuan, Ming; Zhao, Yanxiu; Guo, Yan

    2016-01-01

    The opening and closing of stomata are crucial for plant photosynthesis and transpiration. Actin filaments undergo dynamic reorganization during stomatal closure, but the underlying mechanism for this cytoskeletal reorganization remains largely unclear. In this study, we identified and characterized Arabidopsis thaliana casein kinase 1-like protein 2 (CKL2), which responds to abscisic acid (ABA) treatment and participates in ABA- and drought-induced stomatal closure. Although CKL2 does not bind to actin filaments directly and has no effect on actin assembly in vitro, it colocalizes with and stabilizes actin filaments in guard cells. Further investigation revealed that CKL2 physically interacts with and phosphorylates actin depolymerizing factor 4 (ADF4) and inhibits its activity in actin filament disassembly. During ABA-induced stomatal closure, deletion of CKL2 in Arabidopsis alters actin reorganization in stomata and renders stomatal closure less sensitive to ABA, whereas deletion of ADF4 impairs the disassembly of actin filaments and causes stomatal closure to be more sensitive to ABA. Deletion of ADF4 in the ckl2 mutant partially recues its ABA-insensitive stomatal closure phenotype. Moreover, Arabidopsis ADFs from subclass I are targets of CKL2 in vitro. Thus, our results suggest that CKL2 regulates actin filament reorganization and stomatal closure mainly through phosphorylation of ADF. PMID:27268429

  15. Regulation of the intermediate filament protein nestin at rodent neuromuscular junctions by innervation and activity.

    PubMed

    Kang, Hyuno; Tian, Le; Son, Young-Jin; Zuo, Yi; Procaccino, Diane; Love, Flora; Hayworth, Christopher; Trachtenberg, Joshua; Mikesh, Michelle; Sutton, Lee; Ponomareva, Olga; Mignone, John; Enikolopov, Grigori; Rimer, Mendell; Thompson, Wesley

    2007-05-30

    The intermediate filament nestin is localized postsynaptically at rodent neuromuscular junctions. The protein forms a filamentous network beneath and between the synaptic gutters, surrounds myofiber nuclei, and is associated with Z-discs adjacent to the junction. In situ hybridization shows that nestin mRNA is synthesized selectively by synaptic myonuclei. Although weak immunoreactivity is present in myelinating Schwann cells that wrap the preterminal axon, nestin is not detected in the terminal Schwann cells (tSCs) that cover the nerve terminal branches. However, after denervation of muscle, nestin is upregulated in tSCs and in SCs within the nerve distal to the lesion site. In contrast, immunoreactivity is strongly downregulated in the muscle fiber. Transgenic mice in which the nestin neural enhancer drives expression of a green fluorescent protein (GFP) reporter show that the regulation in SCs is transcriptional. However, the postsynaptic expression occurs through enhancer elements distinct from those responsible for regulation in SCs. Application of botulinum toxin shows that the upregulation in tSCs and the loss of immunoreactivity in muscle fibers occurs with blockade of transmitter release. Extrinsic stimulation of denervated muscle maintains the postsynaptic expression of nestin but does not affect the upregulation in SCs. Thus, a nestin-containing cytoskeleton is promoted in the postsynaptic muscle fiber by nerve-evoked muscle activity but suppressed in tSCs by transmitter release. Nestin antibodies and GFP driven by nestin promoter elements serve as excellent markers for the reactive state of SCs. Vital imaging of GFP shows that SCs grow a dynamic set of processes after denervation.

  16. The Great Solar Active Region NOAA 12192: Helicity Transport, Filament Formation, and Impact on the Polar Field

    NASA Astrophysics Data System (ADS)

    McMaken, Tyler C.; Petrie, Gordon J. D.

    2017-05-01

    The solar active region (AR), NOAA 12192, appeared in 2014 October as the largest AR in 24 years. Here we examine the counterintuitive nature of two diffusion-driven processes in the region: the role of helicity buildup in the formation of a major filament, and the relationship between the effects of supergranular diffusion and meridional flow on the AR and on the polar field. Quantitatively, calculations of current helicity and magnetic twist from Helioseismic and Magnetic Imager (HMI) vector magnetograms indicate that, though AR 12192 emerged with negative helicity, positive helicity from subsequent flux emergence, consistent with the hemispheric sign-preference of helicity, increased over time within large-scale, weak-field regions such as those near the polarity inversion line (PIL). Morphologically, Atmospheric Imaging Assembly observations of filament barbs, sigmoidal patterns, and bases of Fe xii stalks initially exhibited signatures of negative helicity, and the long filament that subsequently formed had a strong positive helicity consistent with the helicity buildup along the PIL. We find from full-disk HMI magnetograms that AR 12192's leading positive flux was initially closer to the equator but, owing either to the region’s magnetic surroundings or to its asymmetric flux density distribution, was transported poleward more quickly on average than its trailing negative flux, contrary to the canonical pattern of bipole flux transport. This behavior caused the AR to have a smaller effect on the polar fields than expected and enabled the formation of the very long neutral line where the filament formed.

  17. The Great Solar Active Region NOAA 12192: Helicity Transport, Filament Formation, and Impact on the Polar Field

    NASA Astrophysics Data System (ADS)

    Petrie, Gordon; McMaken, Tyler C.

    2017-08-01

    The solar active region (AR), NOAA 12192, appeared in 2014 October as the largest AR in 24 years. Here we examine the counterintuitive nature of two diffusion-driven processes in the region: the role of helicity buildup in the formation of a major filament, and the relationship between the effects of supergranular diffusion and meridional flow on the AR and on the polar field. Quantitatively, calculations of current helicity and magnetic twist from Helioseismic and Magnetic Imager (HMI) vector magnetograms indicate that, though AR 12192 emerged with negative helicity, positive helicity from subsequent flux emergence, consistent with the hemispheric sign-preference of helicity, increased over time within large-scale, weak-field regions such as those near the polarity inversion line (PIL). Morphologically, Atmospheric Imaging Assembly observations of filament barbs, sigmoidal patterns, and bases of Fe xii stalks initially exhibited signatures of negative helicity, and the long filament that subsequently formed had a strong positive helicity consistent with the helicity buildup along the PIL. We find from full-disk HMI magnetograms that AR 12192's leading positive flux was initially closer to the equator but, owing either to the region’s magnetic surroundings or to its asymmetric flux density distribution, was transported poleward more quickly on average than its trailing negative flux, contrary to the canonical pattern of bipole flux transport. This behavior caused the AR to have a smaller effect on the polar fields than expected and enabled the formation of the very long neutral line where the filament formed.

  18. A numerical study of multi filament formation in metal-ion based CBRAM

    NASA Astrophysics Data System (ADS)

    Berco, Dan; Tseng, Tseung-Yuen

    2016-02-01

    This study investigates the underlying mechanisms of multiple conductive filaments (CF) creation in metal-ion based conductive bridge RRAM (CBRAM) by using the Metropolis Monte Carlo algorithm and suggests a possible explanation for this phenomenon. The simulation method is demonstrated over a Cu/HfO2 structure, starting from a random initial distribution of oxygen vacancies (OV) defects in the resistive switching layer, to a formed CF and ending in a ruptured state. the results indicate that "Hot Spots" (HS), where agglomeration of OV trap like states for electron hopping based conduction induce local heating, create favorable energy conditions to attract diffused metal species originating from the top electrode. While HS may be created and annihilated by random OV generation and recombination processes, the precipitated metal forms a stem out of which a CF could evolve. The CF stem's final growth stage is mainly driven by drift and diffusion. This process may lead to the formation of one or more CFs as a function of the forming bias voltage. This bias dependence is demonstrated over a large range, where the creation of a single, double and multiple CFs are shown. In addition, the reset process of the multi CF device is presented, and the experimentally observed, step like, gradual CBRAM reset is verified. The simulated results are in good agreement with experimental data and promote the idea that OV defect engineering may be used to improve CBRAM performance.

  19. In Vitro Analysis of Finasteride Activity against Candida albicans Urinary Biofilm Formation and Filamentation

    PubMed Central

    Chavez-Dozal, Alba A.; Lown, Livia; Jahng, Maximillian; Walraven, Carla J.

    2014-01-01

    Candida albicans is the 3rd most common cause of catheter-associated urinary tract infections, with a strong propensity to form drug-resistant catheter-related biofilms. Due to the limited efficacy of available antifungals against biofilms, drug repurposing has been investigated in order to identify novel agents with activities against fungal biofilms. Finasteride is a 5-α-reductase inhibitor commonly used for the treatment of benign prostatic hyperplasia, with activity against human type II and III isoenzymes. We analyzed the Candida Genome Database and identified a C. albicans homolog of type III 5-α-reductase, Dfg10p, which shares 27% sequence identity and 41% similarity to the human type III 5-α-reductase. Thus, we investigated finasteride for activity against C. albicans urinary biofilms, alone and in combination with amphotericin B or fluconazole. Finasteride alone was highly effective in the prevention of C. albicans biofilm formation at doses of ≥16 mg/liter and the treatment of preformed biofilms at doses of ≥128 mg/liter. In biofilm checkerboard analyses, finasteride exhibited synergistic activity in the prevention of biofilm formation in a combination of 4 mg/liter finasteride with 2 mg/liter fluconazole. Finasteride inhibited filamentation, thus suggesting a potential mechanism of action. These results indicate that finasteride alone is highly active in the prevention of C. albicans urinary biofilms in vitro and has synergistic activity in combination with fluconazole. Further investigation of the clinical utility of finasteride in the prevention of urinary candidiasis is warranted. PMID:25049253

  20. In vitro analysis of finasteride activity against Candida albicans urinary biofilm formation and filamentation.

    PubMed

    Chavez-Dozal, Alba A; Lown, Livia; Jahng, Maximillian; Walraven, Carla J; Lee, Samuel A

    2014-10-01

    Candida albicans is the 3rd most common cause of catheter-associated urinary tract infections, with a strong propensity to form drug-resistant catheter-related biofilms. Due to the limited efficacy of available antifungals against biofilms, drug repurposing has been investigated in order to identify novel agents with activities against fungal biofilms. Finasteride is a 5-α-reductase inhibitor commonly used for the treatment of benign prostatic hyperplasia, with activity against human type II and III isoenzymes. We analyzed the Candida Genome Database and identified a C. albicans homolog of type III 5-α-reductase, Dfg10p, which shares 27% sequence identity and 41% similarity to the human type III 5-α-reductase. Thus, we investigated finasteride for activity against C. albicans urinary biofilms, alone and in combination with amphotericin B or fluconazole. Finasteride alone was highly effective in the prevention of C. albicans biofilm formation at doses of ≥16 mg/liter and the treatment of preformed biofilms at doses of ≥128 mg/liter. In biofilm checkerboard analyses, finasteride exhibited synergistic activity in the prevention of biofilm formation in a combination of 4 mg/liter finasteride with 2 mg/liter fluconazole. Finasteride inhibited filamentation, thus suggesting a potential mechanism of action. These results indicate that finasteride alone is highly active in the prevention of C. albicans urinary biofilms in vitro and has synergistic activity in combination with fluconazole. Further investigation of the clinical utility of finasteride in the prevention of urinary candidiasis is warranted.

  1. The Role of Candida albicans SPT20 in Filamentation, Biofilm Formation and Pathogenesis

    PubMed Central

    Tan, Xiaojiang; Fuchs, Beth Burgwyn; Wang, Yan; Chen, Weiping; J. Yuen, Grace; Chen, Rosalyn B.; Jayamani, Elamparithi; Anastassopoulou, Cleo; Pukkila-Worley, Read; Coleman, Jeffrey J.; Mylonakis, Eleftherios

    2014-01-01

    Candida albicans is a ubiquitous fungus, which can cause very serious and sometimes life-threatening infections in susceptible patients. We used Caenorhabditis elegans as a model host to screen a library of C. albicans mutants for decreased virulence and identified SPT20 as important for virulence. The transcription co-activator SPT20 was identified originally as a suppressor of Ty and solo δ insertion mutations, which can cause transcription defects in Saccharomyces cerevisiae. It is resistant to the toxicity caused by overexpression of GAL4-VP16. We constructed a C. albicans spt20Δ/Δ mutant and found the spt20Δ/Δ strain was significantly less virulent than the wild-type strain SC5314 in C. elegans (p < 0.0001), Galleria mellonella (p < 0.01) and mice (p < 0.001). Morphologically, spt20Δ/Δ mutant cells demonstrated a “snow-flake” shape and clustered together; prolonged culture times resulted in increased size of the cluster. The clustered morphology was associated with defects in nuclei distribution, as the nuclei were not observed in many cellular compartments. In addition, the C. albicans spt20Δ/Δ mutant resulted in defects in hyphae and biofilm formation (compared to the wild-type strain, p < 0.05), and sensitivity to cell wall and osmotic stressors, and to antifungal agents. Thus our study demonstrated a role of C. albicans SPT20 in overall morphology and distribution of nuclear material, which may cause the defects in filamentation and biofilm formation directly when this gene is deleted. PMID:24732310

  2. Ubiquitination and filamentous structure of cytidine triphosphate synthase

    PubMed Central

    Pai, Li-Mei; Wang, Pei-Yu; Lin, Wei-Cheng; Chakraborty, Archan; Yeh, Chau-Ting; Lin, Yu-Hung

    2016-01-01

    ABSTRACT Living organisms respond to nutrient availability by regulating the activity of metabolic enzymes. Therefore, the reversible post-translational modification of an enzyme is a common regulatory mechanism for energy conservation. Recently, cytidine-5′-triphosphate (CTP) synthase was discovered to form a filamentous structure that is evolutionarily conserved from flies to humans. Interestingly, induction of the formation of CTP synthase filament is responsive to starvation or glutamine depletion. However, the biological roles of this structure remain elusive. We have recently shown that ubiquitination regulates CTP synthase activity by promoting filament formation in Drosophila ovaries during endocycles. Intriguingly, although the ubiquitination process was required for filament formation induced by glutamine depletion, CTP synthase ubiquitination was found to be inversely correlated with filament formation in Drosophila and human cell lines. In this article, we discuss the putative dual roles of ubiquitination, as well as its physiological implications, in the regulation of CTP synthase structure. PMID:27116391

  3. Correlation of expression of the actin filament-bundling protein espin with stereociliary bundle formation in the developing inner ear.

    PubMed

    Li, Huawei; Liu, Hong; Balt, Steve; Mann, Sabine; Corrales, C Eduardo; Heller, Stefan

    2004-01-01

    The vertebrate hair cell is named for its stereociliary bundle or hair bundle that protrudes from the cell's apical surface. Hair bundles mediate mechanosensitivity, and their highly organized structure plays a critical role in mechanoelectrical transduction and amplification. The prototypical hair bundle is composed of individual stereocilia, 50-300 in number, depending on the animal species and on the type of hair cell. The assembly of stereocilia, in particular, the formation during development of individual rows of stereocilia with descending length, has been analyzed in great morphological detail. Electron microscopic studies have demonstrated that stereocilia are filled with actin filaments that are rigidly cross-linked. The growth of individual rows of stereocilia is associated with the addition of actin filaments and with progressively increasing numbers of cross-bridges between actin filaments. Recently, a mutation in the actin filament-bundling protein espin has been shown to underlie hair bundle degeneration in the deaf jerker mouse, subsequently leading to deafness. Our study was undertaken to investigate the appearance and developmental expression of espin in chicken inner ear sensory epithelia. We found that the onset of espin expression correlates with the initiation and growth of stereocilia bundles in vestibular and cochlear hair cells. Intense espin immunolabeling of stereocilia was colocalized with actin filament staining in all types of hair cells at all developmental stages and in adult animals. Our analysis of espin as a molecular marker for actin filament cross-links in stereocilia is in full accordance with previous morphological studies and implicates espin as an important structural component of hair bundles from initiation of bundle assembly to mature chicken hair cells.

  4. Cross-species microarray hybridization to identify developmentally regulated genes in the filamentous fungus Sordaria macrospora.

    PubMed

    Nowrousian, Minou; Ringelberg, Carol; Dunlap, Jay C; Loros, Jennifer J; Kück, Ulrich

    2005-04-01

    The filamentous fungus Sordaria macrospora forms complex three-dimensional fruiting bodies that protect the developing ascospores and ensure their proper discharge. Several regulatory genes essential for fruiting body development were previously isolated by complementation of the sterile mutants pro1, pro11 and pro22. To establish the genetic relationships between these genes and to identify downstream targets, we have conducted cross-species microarray hybridizations using cDNA arrays derived from the closely related fungus Neurospora crassa and RNA probes prepared from wild-type S. macrospora and the three developmental mutants. Of the 1,420 genes which gave a signal with the probes from all the strains used, 172 (12%) were regulated differently in at least one of the three mutants compared to the wild type, and 17 (1.2%) were regulated differently in all three mutant strains. Microarray data were verified by Northern analysis or quantitative real time PCR. Among the genes that are up- or down-regulated in the mutant strains are genes encoding the pheromone precursors, enzymes involved in melanin biosynthesis and a lectin-like protein. Analysis of gene expression in double mutants revealed a complex network of interaction between the pro gene products.

  5. Filament formation and erasure in molybdenum oxide during resistive switching cycles

    NASA Astrophysics Data System (ADS)

    Kudo, Masaki; Arita, Masashi; Ohno, Yuuki; Takahashi, Yasuo

    2014-10-01

    In-situ filament observations were carried out on the Cu/MoOx/TiN resistive random access memory (ReRAM) by using transmission electron microscopy. Multiple positive and negative I-V cycles were investigated. Clear set-reset bipolar switch corresponding to the characteristics of conventional ReRAM devices was recognized. Filament grew from TiN to Cu in the set cycle and shrank from TiN to Cu in the reset cycle. However, there was no clear contrast change in the image at the switching moment, and thus, switching is thought to occur in a local region of the filament. When the current was large at reset, the filament shrank much, and its position tended to change.

  6. Filament formation and erasure in molybdenum oxide during resistive switching cycles

    SciTech Connect

    Kudo, Masaki; Arita, Masashi Ohno, Yuuki; Takahashi, Yasuo

    2014-10-27

    In-situ filament observations were carried out on the Cu/MoO{sub x}/TiN resistive random access memory (ReRAM) by using transmission electron microscopy. Multiple positive and negative I-V cycles were investigated. Clear set-reset bipolar switch corresponding to the characteristics of conventional ReRAM devices was recognized. Filament grew from TiN to Cu in the set cycle and shrank from TiN to Cu in the reset cycle. However, there was no clear contrast change in the image at the switching moment, and thus, switching is thought to occur in a local region of the filament. When the current was large at reset, the filament shrank much, and its position tended to change.

  7. Formation of actin filament bundles in the ring canals of developing Drosophila follicles

    PubMed Central

    1996-01-01

    Growing the intracellular bridges that connect nurse cells with each o ther and to the developing oocyte is vital for egg development. These ring canals increase from 0.5 microns in diameter at stage 2 to 10 microns in diameter at stage 11. Thin sections cut horizontally as you would cut a bagel, show that there is a layer of circumferentially oriented actin filaments attached to the plasma membrane at the periphery of each canal. By decoration with subfragment 1 of myosin we find actin filaments of mixed polarities in the ring such as found in the "contractile ring" formed during cytokinesis. In vertical sections through the canal the actin filaments appear as dense dots. At stage 2 there are 82 actin filaments in the ring, by stage 6 there are 717 and by stage 10 there are 726. Taking into account the diameter, this indicates that there is 170 microns of actin filaments/canal at stage 2 (pi x 0.5 microns x 82), 14,000 microns at stage 9 and approximately 23,000 microns at stage 11 or one inch of actin filament! The density of actin filaments remains unchanged throughout development. What is particularly striking is that by stages 4-5, the ring of actin filaments has achieved its maximum thickness, even though the diameter has not yet increased significantly. Thereafter, the diameter increases. Throughout development, stages 2-11, the canal length also increases. Although the density (number of actin filaments/micron2) through a canal remains constant from stage 5 on, the actin filaments appear as a net of interconnected bundles. Further information on this net of bundles comes from studying mutant animals that lack kelch, a protein located in the ring canal that has homology to the actin binding protein, scruin. In this mutant, the actin filaments form normally but individual bundles that comprise the fibers of the net are not bound tightly together. Some bundles enter into the ring canal lumen but do not completely occlude the lumen. all these observations lay

  8. Candida albicans OPI1 Regulates Filamentous Growth and Virulence in Vaginal Infections, but Not Inositol Biosynthesis

    PubMed Central

    Chen, Ying-Lien; de Bernardis, Flavia; Yu, Shang-Jie; Sandini, Silvia; Kauffman, Sarah; Tams, Robert N.; Bethea, Emily; Reynolds, Todd B.

    2015-01-01

    ScOpi1p is a well-characterized transcriptional repressor and master regulator of inositol and phospholipid biosynthetic genes in the baker’s yeast Saccharomyces cerevisiae. An ortholog has been shown to perform a similar function in the pathogenic fungus Candida glabrata, but with the distinction that CgOpi1p is essential for growth in this organism. However, in the more distantly related yeast Yarrowia lipolytica, the OPI1 homolog was not found to regulate inositol biosynthesis, but alkane oxidation. In Candida albicans, the most common cause of human candidiasis, its Opi1p homolog, CaOpi1p, has been shown to complement a S. cerevisiae opi1∆ mutant for inositol biosynthesis regulation when heterologously expressed, suggesting it might serve a similar role in this pathogen. This was tested in the pathogen directly in this report by disrupting the OPI1 homolog and examining its phenotypes. It was discovered that the OPI1 homolog does not regulate INO1 expression in C. albicans, but it does control SAP2 expression in response to bovine serum albumin containing media. Meanwhile, we found that CaOpi1 represses filamentous growth at lower temperatures (30°C) on agar, but not in liquid media. Although, the mutant does not affect virulence in a mouse model of systemic infection, it does affect virulence in a rat model of vaginitis. This may be because Opi1p regulates expression of the SAP2 protease, which is required for rat vaginal infections. PMID:25602740

  9. Regulation of the actin cycle in vivo by actin filament severing

    PubMed Central

    McGrath, James L.; Osborn, Eric A.; Tardy, Yanik S.; Dewey, C. Forbes; Hartwig, John H.

    2000-01-01

    Cycling of actin subunits between monomeric and filamentous phases is essential for cell crawling behavior. We investigated actin filament turnover rates, length, number, barbed end exposure, and binding of cofilin in bovine arterial endothelial cells moving at different speeds depending on their position in a confluent monolayer. Fast-translocating cells near the wound edge have short filament lifetimes compared with turnover values that proportionately increase in slower moving cells situated at increasing distances from the wound border. Contrasted with slow cells exhibiting slow actin filament turnover speeds, fast cells have less polymerized actin, shorter actin filaments, more free barbed ends, and less actin-associated cofilin. Cultured primary fibroblasts manifest identical relationships between speed and actin turnover as the endothelial cells, and fast fibroblasts expressing gelsolin have higher actin turnover rates than slow fibroblasts that lack this actin-severing protein. These results implicate actin filament severing as an important control mechanism for actin cycling in cells. PMID:10823888

  10. The zinc-finger transcription factor, Ofi1, regulates white-opaque switching and filamentation in the yeast Candida albicans.

    PubMed

    Du, Han; Li, Xiaoling; Huang, Guanghua; Kang, Yingqian; Zhu, Liquan

    2015-05-01

    Candida albicans is a major fungal pathogen of humans. The most striking biological feature of C. albicans is its phenotypic plasticity, allowing it to undergo morphological transitions in response to various environmental cues. Transcription factors play critical roles in the regulation of morphological transitions. Here, we report the role of opaque and filamentation inducer 1 (Ofi1), a previously uncharacterized zinc-finger-containing protein encoded by the gene orf19.4972, in the regulation of white-opaque switching and filamentous growth. Over-expression of OFI1 not only induced white-to-opaque switching but also promoted filamentation and invasive growth in C. albicans. Deletion of OFI1 had no obvious effect on filamentation under the culture conditions tested, while deletion of OFI1 reduced the frequency of white-to-opaque switching. We propose that Ofi1 functions downstream of Wor1, the master regulator of white-opaque switching. However, over-expression of OFI1 in the wor1/wor1 mutant could not induce the opaque phenotype, suggesting that Ofi1 does not work alone and other transcription factors downstream of Wor1 are also involved in this regulation. Given the importance of Ofi1 in the regulation of white-opaque switching and filamentation, the present study establishes a new link between these two processes. © The Author 2015. Published by ABBS Editorial Office in association with Oxford University Press on behalf of the Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences.

  11. Isolation and characterization of Candida albicans morphological mutants derepressed for the formation of filamentous hypha-type structures

    SciTech Connect

    Gil, C.; Pomes, R.; Nombela, C. )

    1990-05-01

    Several Candida albicans morphological mutants were obtained by a procedure based on a combined treatment with nitrous acid plus UV irradiation and a double-enrichment step to increase the proportion of mutants growing as long filamentous structures. Altered cell morphogenesis in these mutants correlated with an altered colonial phenotype. Two of these mutants, C. albicans NEL102 and NEL103, were selected and characterized. Mutant blastoconidia initiated budding but eventually gave rise to filamentous hypha-type formations. These filaments were long and septate, and they branched very regularly at positions near septa. Calcofluor white (which is known to bind chitin-rich areas) stained septa, branching zones, and filament tips very intensely, as observed under the fluorescence microscope. Wild-type hybrids were obtained by fusing protoplasts of strain NEL102 with B14, another morphological mutant previously described as being permanently pseudomycelial, indicating that genetic determinants responsible for the two altered phenotypes are different. The mutants characterized in this work seemed to sequentially express the morphogenic characteristics of C. albicans, from blastoconidia to hyphae, in the absence of any inducer. Further characterization of these strains could be relevant to gain understanding of the genetic control of dimorphism in this species.

  12. Actin Filament Polymerization Regulates Gliding Motility by Apicomplexan ParasitesV⃞

    PubMed Central

    Wetzel, D.M.; Håkansson, S.; Hu, K.; Roos, D.; Sibley, L.D.

    2003-01-01

    Host cell entry by Toxoplasma gondii depends critically on actin filaments in the parasite, yet paradoxically, its actin is almost exclusively monomeric. In contrast to the absence of stable filaments in conventional samples, rapid-freeze electron microscopy revealed that actin filaments were formed beneath the plasma membrane of gliding parasites. To investigate the role of actin filaments in motility, we treated parasites with the filament-stabilizing drug jasplakinolide (JAS) and monitored the distribution of actin in live and fixed cells using yellow fluorescent protein (YFP)-actin. JAS treatment caused YFP-actin to redistribute to the apical and posterior ends, where filaments formed a spiral pattern subtending the plasma membrane. Although previous studies have suggested that JAS induces rigor, videomicroscopy demonstrated that JAS treatment increased the rate of parasite gliding by approximately threefold, indicating that filaments are rate limiting for motility. However, JAS also frequently reversed the normal direction of motility, disrupting forward migration and cell entry. Consistent with this alteration, subcortical filaments in JAS-treated parasites occurred in tangled plaques as opposed to the straight, roughly parallel orientation observed in control cells. These studies reveal that precisely controlled polymerization of actin filaments imparts the correct timing, duration, and directionality of gliding motility in the Apicomplexa. PMID:12589042

  13. Dwarf Galaxy Formation with H2-regulated Star Formation

    NASA Astrophysics Data System (ADS)

    Kuhlen, Michael; Krumholz, Mark R.; Madau, Piero; Smith, Britton D.; Wise, John

    2012-04-01

    We describe cosmological galaxy formation simulations with the adaptive mesh refinement code Enzo that incorporate a star formation prescription regulated by the local abundance of molecular hydrogen. We show that this H2-regulated prescription leads to a suppression of star formation in low-mass halos (Mh <~ 1010 M ⊙) at z > 4, alleviating some of the dwarf galaxy problems faced by theoretical galaxy formation models. H2 regulation modifies the efficiency of star formation of cold gas directly, rather than indirectly reducing the cold gas content with "supernova feedback." We determine the local H2 abundance in our most refined grid cells (76 proper parsec in size at z = 4) by applying the model of Krumholz, McKee, & Tumlinson, which is based on idealized one-dimensional radiative transfer calculations of H2 formation-dissociation balance in ~100 pc atomic-molecular complexes. Our H2-regulated simulations are able to reproduce the empirical (albeit lower z) Kennicutt-Schmidt relation, including the low Σgas cutoff due to the transition from atomic to molecular phase and the metallicity dependence thereof, without the use of an explicit density threshold in our star formation prescription. We compare the evolution of the luminosity function, stellar mass density, and star formation rate density from our simulations to recent observational determinations of the same at z = 4-8 and find reasonable agreement between the two.

  14. Multiple roles for keratin intermediate filaments in the regulation of epithelial barrier function and apico-basal polarity

    PubMed Central

    Salas, Pedro J.; Forteza, Radia; Mashukova, Anastasia

    2016-01-01

    abstract As multicellular organisms evolved a family of cytoskeletal proteins, the keratins (types I and II) expressed in epithelial cells diversified in more than 20 genes in vertebrates. There is no question that keratin filaments confer mechanical stiffness to cells. However, such a number of genes can hardly be explained by evolutionary advantages in mechanical features. The use of transgenic mouse models has revealed unexpected functional relationships between keratin intermediate filaments and intracellular signaling. Accordingly, loss of keratins or mutations in keratins that cause or predispose to human diseases, result in increased sensitivity to apoptosis, regulation of innate immunity, permeabilization of tight junctions, and mistargeting of apical proteins in different epithelia. Precise mechanistic explanations for these phenomena are still lacking. However, immobilization of membrane or cytoplasmic proteins, including chaperones, on intermediate filaments (“scaffolding”) appear as common molecular mechanisms and may explain the need for so many different keratin genes in vertebrates. PMID:27583190

  15. Emerging roles of sumoylation in the regulation of actin, microtubules, intermediate filaments, and septins

    PubMed Central

    Alonso, Annabel; Greenlee, Matt; Matts, Jessica; Kline, Jake; Davis, Kayla J.

    2015-01-01

    Sumoylation is a powerful regulatory system that controls many of the critical processes in the cell, including DNA repair, transcriptional regulation, nuclear transport, and DNA replication. Recently, new functions for SUMO have begun to emerge. SUMO is covalently attached to components of each of the four major cytoskeletal networks, including microtubule‐associated proteins, septins, and intermediate filaments, in addition to nuclear actin and actin‐regulatory proteins. However, knowledge of the mechanisms by which this signal transduction system controls the cytoskeleton is still in its infancy. One story that is beginning to unfold is that SUMO may regulate the microtubule motor protein dynein by modification of its adaptor Lis1. In other instances, cytoskeletal elements can both bind to SUMO non‐covalently and also be conjugated by it. The molecular mechanisms for many of these new functions are not yet clear, but are under active investigation. One emerging model links the function of MAP sumoylation to protein degradation through SUMO‐targeted ubiquitin ligases, also known as STUbL enzymes. Other possible functions for cytoskeletal sumoylation are also discussed. © 2015 The Authors. Cytoskeleton Published by Wiley Periodicals, Inc. PMID:26033929

  16. Radio-Optical Alignment and Recent Star Formation Associated with Ionized Filaments in the Halo of NGC 5128 (Centaurus A)

    NASA Astrophysics Data System (ADS)

    Rejkuba, M.; Minniti, D.; Courbin, F.; Silva, D. R.

    2002-01-01

    We used a direct CCD camera at the Magellan I telescope at Las Campanas Observatory and the Focal Reducer/Low Dispersion Spectrograph (FORS1) at the Antu Very Large Telescope (VLT) ESO Paranal Observatory to image fields centered on the inner and outer optical filaments in the halo of NGC 5128. In the V versus U-V color-magnitude diagrams we have identified young blue supergiants associated with these line-emitting filaments located between the inner radio lobe and the northern middle lobe. Around the outer filament, stars as young as 10 Myr were detected. They are principally aligned with the direction of the radio jet, but a vertical north-south alignment along the edge of the H I cloud is also present. Young stars in the inner filament field are found inside the bright knots of photoionized gas and are strongly aligned in the direction of the center of the galaxy at the same position angle as the inner radio jet. Fitting the Padova isochrones on UV color-magnitude diagrams, we find that blue stars around the inner filaments have ages similar to the ones around the outer filaments ~10-15 Myr and the same abundance of Z=0.004. The presence of young blue supergiants clearly shows that the bright blue knots in the northeastern halo of NGC 5128 are associations of young stars with photoionized gas. The temperature of the brightest stars is T~12,000-16,000 K, insufficient to account alone for the high excitation lines observed in the surrounding ionized gas. Thus, the optical emission jet is principally seen due to its alignment with the radio structure of the active galactic nucleus (AGN). The highly collimated star formation is present only in the northeastern halo of the galaxy, suggesting interaction of the jet with the gas clouds deposited during the last accretion event as the preferred triggering mechanism. From these observations, we infer a lower limit for the age of the NGC 5128 jet at 107 yr. The triggering of the star formation in the dense clouds in the

  17. Dynamic Filament Formation by a Divergent Bacterial Actin-Like ParM Protein

    PubMed Central

    Brzoska, Anthony J.; Jensen, Slade O.; Barton, Deborah A.; Davies, Danielle S.; Overall, Robyn L.; Skurray, Ronald A.; Firth, Neville

    2016-01-01

    Actin-like proteins (Alps) are a diverse family of proteins whose genes are abundant in the chromosomes and mobile genetic elements of many bacteria. The low-copy-number staphylococcal multiresistance plasmid pSK41 encodes ParM, an Alp involved in efficient plasmid partitioning. pSK41 ParM has previously been shown to form filaments in vitro that are structurally dissimilar to those formed by other bacterial Alps. The mechanistic implications of these differences are not known. In order to gain insights into the properties and behavior of the pSK41 ParM Alp in vivo, we reconstituted the parMRC system in the ectopic rod-shaped host, E. coli, which is larger and more genetically amenable than the native host, Staphylococcus aureus. Fluorescence microscopy showed a functional fusion protein, ParM-YFP, formed straight filaments in vivo when expressed in isolation. Strikingly, however, in the presence of ParR and parC, ParM-YFP adopted a dramatically different structure, instead forming axial curved filaments. Time-lapse imaging and selective photobleaching experiments revealed that, in the presence of all components of the parMRC system, ParM-YFP filaments were dynamic in nature. Finally, molecular dissection of the parMRC operon revealed that all components of the system are essential for the generation of dynamic filaments. PMID:27310470

  18. Temperature regulation of gliding motility in filamentous sulfur bacteria, Beggiatoa spp.

    PubMed

    Dunker, Rita; Røy, Hans; Jørgensen, Bo Barker

    2010-08-01

    The response of gliding motility to changing temperatures was studied in filaments of the large sulfur bacteria Beggiatoa from arctic, temperate and tropical marine environments. The general shape of the gliding speed vs. temperature curves from all three locations was similar, but differed in the maximal gliding speed of the filaments, optimum temperature and the temperature range of motility. The optimum temperature and the overall temperature range of gliding motility accorded to the climatic origin of the filaments with a high temperature range for tropical, an intermediate range for temperate, and a low temperature range for arctic filaments. The temperature-controlled decrease in gliding speed at low temperatures was reversible while the decline in speed at high temperatures was due to irreversible thermal damage in individual filaments. Filaments from the Arctic and cold-acclimatized filaments from the temperate zone were unaffected by transient freezing of the surrounding seawater. At in situ temperatures, filaments glided at 17-55% of the gliding speed at the optimum temperatures, indicating that they were well adapted to the temperature regime of their origin. Our results point towards an enzymatic control of temperature-dependent gliding motility.

  19. The Peptidoglycan-Binding Protein SjcF1 Influences Septal Junction Function and Channel Formation in the Filamentous Cyanobacterium Anabaena.

    PubMed

    Rudolf, Mareike; Tetik, Nalan; Ramos-León, Félix; Flinner, Nadine; Ngo, Giang; Stevanovic, Mara; Burnat, Mireia; Pernil, Rafael; Flores, Enrique; Schleiff, Enrico

    2015-06-30

    Filamentous, heterocyst-forming cyanobacteria exchange nutrients and regulators between cells for diazotrophic growth. Two alternative modes of exchange have been discussed involving transport either through the periplasm or through septal junctions linking adjacent cells. Septal junctions and channels in the septal peptidoglycan are likely filled with septal junction complexes. While possible proteinaceous factors involved in septal junction formation, SepJ (FraG), FraC, and FraD, have been identified, little is known about peptidoglycan channel formation and septal junction complex anchoring to the peptidoglycan. We describe a factor, SjcF1, involved in regulation of septal junction channel formation in the heterocyst-forming cyanobacterium Anabaena sp. strain PCC 7120. SjcF1 interacts with the peptidoglycan layer through two peptidoglycan-binding domains and is localized throughout the cell periphery but at higher levels in the intercellular septa. A strain with an insertion in sjcF1 was not affected in peptidoglycan synthesis but showed an altered morphology of the septal peptidoglycan channels, which were significantly wider in the mutant than in the wild type. The mutant was impaired in intercellular exchange of a fluorescent probe to a similar extent as a sepJ deletion mutant. SjcF1 additionally bears an SH3 domain for protein-protein interactions. SH3 binding domains were identified in SepJ and FraC, and evidence for interaction of SjcF1 with both SepJ and FraC was obtained. SjcF1 represents a novel protein involved in structuring the peptidoglycan layer, which links peptidoglycan channel formation to septal junction complex function in multicellular cyanobacteria. Nonetheless, based on its subcellular distribution, this might not be the only function of SjcF1. Cell-cell communication is central not only for eukaryotic but also for multicellular prokaryotic systems. Principles of intercellular communication are well established for eukaryotes, but the

  20. A cis-acting sequence homologous to the yeast filamentation and invasion response element regulates expression of a pectinase gene from the bean pathogen Colletotrichum lindemuthianum.

    PubMed

    Herbert, Corentin; Jacquet, Christophe; Borel, Charlotte; Esquerre-Tugaye, Marie-Therese; Dumas, Bernard

    2002-08-09

    Phytopathogenic fungi secrete hydrolytic enzymes that degrade plant cell walls, notably pectinases. The signaling pathway(s) that control pectinase gene expression are currently unknown in filamentous fungi. Recently, the green fluorescent protein coding sequence was used as a reporter gene to study the expression of CLPG2, a gene encoding an endopolygalacturonase of the bean pathogen Colletotrichum lindemuthianum. CLPG2 is transcriptionally induced by pectin in the axenic culture of the fungus and during formation of the appressorium, an infection structure specialized in plant tissue penetration. In the present study, promoter deletion and mutagenesis, as well as gel shift mobility assays, allowed for the first time identification of cis-acting elements that bind protein factors and are essential for the regulation of a pectinase gene. We found that two different adjacent DNA motifs are combined to form an active element that shows a strong sequence homology with the yeast filamentation and invasion response element. The same element is required for the transcriptional activation of CLPG2 by pectin and during appressorium development. This study strongly suggests that the control of virulence genes of fungal plant pathogens, such as pectinases, involves the formation of a complex of transcriptional activators similar to those regulating the invasive growth in yeast.

  1. Identification of Fic-1 as an enzyme that inhibits bacterial DNA replication by AMPylating GyrB, promoting filament formation.

    PubMed

    Lu, Canhua; Nakayasu, Ernesto S; Zhang, Li-Qun; Luo, Zhao-Qing

    2016-01-26

    The morphology of bacterial cells is important for virulence, evasion of the host immune system, and coping with environmental stresses. The widely distributed Fic proteins (filamentation induced by cAMP) are annotated as proteins involved in cell division because of the presence of the HPFx[D/E]GN[G/K]R motif. We showed that the presence of Fic-1 from Pseudomonas fluorescens significantly reduced the yield of plasmid DNA when expressed in Escherichia coli or P. fluorescens. Fic-1 interacted with GyrB, a subunit of DNA gyrase, which is essential for bacterial DNA replication. Fic-1 catalyzed the AMPylation of GyrB at Tyr(109), a residue critical for binding ATP, and exhibited auto-AMPylation activity. Mutation of the Fic-1 auto-AMPylated site greatly reduced AMPylation activity toward itself and toward GyrB. Fic-1-dependent AMPylation of GyrB triggered the SOS response, indicative of DNA replication stress or DNA damage. Fic-1 also promoted the formation of elongated cells when the SOS response was blocked. We identified an α-inhibitor protein that we named anti-Fic-1 (AntF), encoded by a gene immediately upstream of Fic-1. AntF interacted with Fic-1, inhibited the AMPylation activity of Fic-1 for GyrB in vitro, and blocked Fic-1-mediated inhibition of DNA replication in bacteria, suggesting that Fic-1 and AntF comprise a toxin-antitoxin module. Our work establishes Fic-1 as an AMPylating enzyme that targets GyrB to inhibit DNA replication and may target other proteins to regulate bacterial morphology.

  2. Regulation of structure and function of sarcomeric actin filaments in striated muscle of the nematode Caenorhabditis elegans

    PubMed Central

    Ono, Shoichiro

    2014-01-01

    The nematode Caenorhabditis elegans has been used as a valuable system to study structure and function of striated muscle. The body wall muscle of C. elegans is obliquely striated muscle with highly organized sarcomeric assembly of actin, myosin, and other accessary proteins. Genetic and molecular biological studies in C. elegans have identified a number of genes encoding structural and regulatory components for the muscle contractile apparatuses, and many of them have counterparts in mammalian cardiac and skeletal muscles or striated muscles in other invertebrates. Applicability of genetics, cell biology, and biochemistry has made C. elegans an excellent system to study mechanisms of muscle contractility and assembly and maintenance of myofibrils. This review focuses on the regulatory mechanisms of structure and function of actin filaments in the C. elegans body wall muscle. Sarcomeric actin filaments in C. elegans muscle are associated with the troponin-tropomyosin system that regulates the actin-myosin interaction. Proteins that bind to the side and ends of actin filaments support ordered assembly of thin filaments. Furthermore, regulators of actin dynamics play important roles in initial assembly, growth, and maintenance of sarcomeres. The knowledge acquired in C. elegans can serve as bases to understand the basic mechanisms of muscle structure and function. PMID:25125169

  3. ATP-dependent regulation of actin monomer-filament equilibrium by cyclase-associated protein and ADF/cofilin.

    PubMed

    Nomura, Kazumi; Ono, Shoichiro

    2013-07-15

    CAP (cyclase-associated protein) is a conserved regulator of actin filament dynamics. In the nematode Caenorhabditis elegans, CAS-1 is an isoform of CAP that is expressed in striated muscle and regulates sarcomeric actin assembly. In the present study, we report that CAS-2, a second CAP isoform in C. elegans, attenuates the actin-monomer-sequestering effect of ADF (actin depolymerizing factor)/cofilin to increase the steady-state levels of actin filaments in an ATP-dependent manner. CAS-2 binds to actin monomers without a strong preference for either ATP- or ADP-actin. CAS-2 strongly enhances the exchange of actin-bound nucleotides even in the presence of UNC-60A, a C. elegans ADF/cofilin that inhibits nucleotide exchange. UNC-60A induces the depolymerization of actin filaments and sequesters actin monomers, whereas CAS-2 reverses the monomer-sequestering effect of UNC-60A in the presence of ATP, but not in the presence of only ADP or the absence of ATP or ADP. A 1:100 molar ratio of CAS-2 to UNC-60A is sufficient to increase actin filaments. CAS-2 has two independent actin-binding sites in its N- and C-terminal halves, and the C-terminal half is necessary and sufficient for the observed activities of the full-length CAS-2. These results suggest that CAS-2 (CAP) and UNC-60A (ADF/cofilin) are important in the ATP-dependent regulation of the actin monomer-filament equilibrium.

  4. THE RAPID FORMATION OF A FILAMENT CAUSED BY MAGNETIC RECONNECTION BETWEEN TWO SETS OF DARK THREADLIKE STRUCTURES

    SciTech Connect

    Yang, Bo; Jiang, Yunchun; Yang, Jiayan; Yu, Shunping; Xu, Zhe E-mail: yjy@ynao.ac.cn

    2016-01-01

    Taking advantage of the high spatiotemporal resolution observations from the Atmospheric Imaging Assembly (AIA) and the Helioseismic and Magnetic Imager on board the Solar Dynamics Observatory, we present rare observations of the rapid formation of a filament caused by magnetic reconnection between two sets of dark threadlike structures. The two sets of dark threadlike structures belong to distinct flux systems with their adjacent ends anchored in an opposite-polarity magnetic field region, where the calculated photospheric velocity field shows that converging flows dominate there. Due to the converging flows, opposite-polarity magnetic flux converged and then canceled, leading to the formation of extreme ultraviolet (EUV) brightening that spread in opposite directions along the spine of the dark threadlike structures. Meanwhile, very weak remote brightening in the other terminals of the dark threadlike structures, as well as EUV loops, which rooted in the opposite-polarity magnetic field region, appeared. In addition, all of the AIA Fe line observations reveal that a flux rope was formed and underwent a rolling motion during the fadeaway of the EUV brightening. Soon after, as the EUV brightening disappeared, a filament that is very likely composed of two sets of intertwined dark threadlike structures was formed. Via differential emission measure (EM) analysis, it is found that both the EM and temperature of the plasma around the flux-canceling site increased during the brightening, implying that there, magnetic reconnection may occur to heat the plasma. These observations provide evidence that the filament is formed by magnetic reconnection associated with flux convergence and cancellation, and the magnetic structure of the filament is most likely a flux rope.

  5. ETOILE Regulates Developmental Patterning in the Filamentous Brown Alga Ectocarpus siliculosus[W

    PubMed Central

    Le Bail, Aude; Billoud, Bernard; Le Panse, Sophie; Chenivesse, Sabine; Charrier, Bénédicte

    2011-01-01

    Brown algae are multicellular marine organisms evolutionarily distant from both metazoans and land plants. The molecular or cellular mechanisms that govern the developmental patterning in brown algae are poorly characterized. Here, we report the first morphogenetic mutant, étoile (etl), produced in the brown algal model Ectocarpus siliculosus. Genetic, cellular, and morphometric analyses showed that a single recessive locus, ETL, regulates cell differentiation: etl cells display thickening of the extracellular matrix (ECM), and the elongated, apical, and actively dividing E cells are underrepresented. As a result of this defect, the overrepresentation of round, branch-initiating R cells in the etl mutant leads to the rapid induction of the branching process at the expense of the uniaxial growth in the primary filament. Computational modeling allowed the simulation of the etl mutant phenotype by including a modified response to the neighborhood information in the division rules used to specify wild-type development. Microarray experiments supported the hypothesis of a defect in cell–cell communication, as primarily Lin-Notch-domain transmembrane proteins, which share similarities with metazoan Notch proteins involved in binary cell differentiation were repressed in etl. Thus, our study highlights the role of the ECM and of novel transmembrane proteins in cell–cell communication during the establishment of the developmental pattern in this brown alga. PMID:21478443

  6. Zyxin regulates endothelial von Willebrand factor secretion by reorganizing actin filaments around exocytic granules

    PubMed Central

    Han, Xiaofan; Li, Pin; Yang, Zhenghao; Huang, Xiaoshuai; Wei, Guoqin; Sun, Yujie; Kang, Xuya; Hu, Xueting; Deng, Qiuping; Chen, Liangyi; He, Aibin; Huo, Yingqing; Li, Dong; Betzig, Eric; Luo, Jincai

    2017-01-01

    Endothelial exocytosis of Weibel–Palade body (WPB) is one of the first lines of defence against vascular injury. However, the mechanisms that control WPB exocytosis in the final stages (including the docking, priming and fusion of granules) are poorly understood. Here we show that the focal adhesion protein zyxin is crucial in this process. Zyxin downregulation inhibits the secretion of von Willebrand factor (VWF), the most abundant cargo in WPBs, from human primary endothelial cells (ECs) induced by cAMP agonists. Zyxin-deficient mice exhibit impaired epinephrine-stimulated VWF release, prolonged bleeding time and thrombosis, largely due to defective endothelial secretion of VWF. Using live-cell super-resolution microscopy, we visualize previously unappreciated reorganization of pre-existing actin filaments around WPBs before fusion, dependent on zyxin and an interaction with the actin crosslinker α-actinin. Our findings identify zyxin as a physiological regulator of endothelial exocytosis through reorganizing local actin network in the final stage of exocytosis. PMID:28256511

  7. ETOILE regulates developmental patterning in the filamentous brown alga Ectocarpus siliculosus.

    PubMed

    Le Bail, Aude; Billoud, Bernard; Le Panse, Sophie; Chenivesse, Sabine; Charrier, Bénédicte

    2011-04-01

    Brown algae are multicellular marine organisms evolutionarily distant from both metazoans and land plants. The molecular or cellular mechanisms that govern the developmental patterning in brown algae are poorly characterized. Here, we report the first morphogenetic mutant, étoile (etl), produced in the brown algal model Ectocarpus siliculosus. Genetic, cellular, and morphometric analyses showed that a single recessive locus, ETL, regulates cell differentiation: etl cells display thickening of the extracellular matrix (ECM), and the elongated, apical, and actively dividing E cells are underrepresented. As a result of this defect, the overrepresentation of round, branch-initiating R cells in the etl mutant leads to the rapid induction of the branching process at the expense of the uniaxial growth in the primary filament. Computational modeling allowed the simulation of the etl mutant phenotype by including a modified response to the neighborhood information in the division rules used to specify wild-type development. Microarray experiments supported the hypothesis of a defect in cell-cell communication, as primarily Lin-Notch-domain transmembrane proteins, which share similarities with metazoan Notch proteins involved in binary cell differentiation were repressed in etl. Thus, our study highlights the role of the ECM and of novel transmembrane proteins in cell-cell communication during the establishment of the developmental pattern in this brown alga.

  8. Chaperonin filaments: The archaeal cytoskeleton?

    PubMed Central

    Trent, Jonathan D.; Kagawa, Hiromi K.; Yaoi, Takuro; Olle, Eric; Zaluzec, Nestor J.

    1997-01-01

    Chaperonins are high molecular mass double-ring structures composed of 60-kDa protein subunits. In the hyperthermophilic archaeon Sulfolobus shibatae the two chaperonin proteins represent ≈4% of its total protein and have a combined intracellular concentration of >30 mg/ml. At concentrations ≥ 0.5 mg/ml purified chaperonins form filaments in the presence of Mg2+ and nucleotides. Filament formation requires nucleotide binding (not hydrolysis), and occurs at physiological temperatures in biologically relevant buffers, including a buffer made from cell extracts. These observations suggest that chaperonin filaments may exist in vivo and the estimated 4600 chaperonins per cell suggest that such filaments could form an extensive cytostructure. We observed filamentous structures in unfixed, uranyl-acetate-stained S. shibatae cells, which resemble the chaperonin filaments in size and appearance. ImmunoGold (Janssen) labeling using chaperonin antibodies indicated that many chaperonins are associated with insoluble cellular structures and these structures appear to be filamentous in some areas, although they could not be uranyl-acetate-stained. The existence of chaperonin filaments in vivo suggests a mechanism whereby their protein-folding activities can be regulated. More generally, the filaments themselves may play a cytoskeletal role in Archaea. PMID:9144246

  9. Temperature change does not affect force between regulated actin filaments and heavy meromyosin in single-molecule experiments.

    PubMed

    Kawai, Masataka; Kido, Takanori; Vogel, Martin; Fink, Rainer H A; Ishiwata, Shin'ichi

    2006-08-01

    The temperature dependence of sliding velocity, force and the number of cross-bridges was studied on regulated actin filaments (reconstituted thin filaments) when they were placed on heavy meromyosin (HMM) attached to a glass surface. The regulated actin filaments were used because our previous study on muscle fibres demonstrated that the temperature effect was much reduced in the absence of regulatory proteins. A fluorescently labelled thin filament was attached to the gelsolin-coated surface of a polystyrene bead. The bead was trapped by optical tweezers, and HMM-thin filament interaction was performed at 20-35 degrees C to study the temperature dependence of force at the single-molecule level. Our experiments showed that there was a small increase in force with temperature (Q10 = 1.43) and sliding velocity (Q10 = 1.46). The small increase in force was correlated with the small increase in the number of cross-bridges (Q10 = 1.49), and when force was divided by the number of cross-bridges, the result did not depend on the temperature (Q(10) = 1.03). These results demonstrate that the force each cross-bridge generates is fixed and independent of temperature. Our additional experiments demonstrate that tropomyosin (Tm) in the presence of troponin (Tn) and Ca2+ enhances both force and velocity, and a truncated mutant, Delta23Tm, diminishes force and velocity. These results are consistent with the hypothesis that Tm in the presence of Tn and Ca2+ exerts a positive allosteric effect on actin to make actomyosin linkage more secure so that larger forces can be generated.

  10. Toxoplasma gondii profilin acts primarily to sequester G-actin while formins efficiently nucleate actin filament formation in vitro.

    PubMed

    Skillman, Kristen M; Daher, Wassim; Ma, Christopher I; Soldati-Favre, Dominique; Sibley, L David

    2012-03-27

    Apicomplexan parasites employ gliding motility that depends on the polymerization of parasite actin filaments for host cell entry. Despite this requirement, parasite actin remains almost entirely unpolymerized at steady state; formation of filaments required for motility relies on a small repertoire of actin-binding proteins. Previous studies have shown that apicomplexan formins and profilin exhibit canonical functions on heterologous actins from higher eukaryotes; however, their biochemical properties on parasite actins are unknown. We therefore analyzed the impact of T. gondii profilin (TgPRF) and FH1-FH2 domains of two formin isoforms in T. gondii (TgFRM1 and TgFRM2) on the polymerization of T. gondii actin (TgACTI). Our findings based on in vitro assays demonstrate that TgFRM1-FH1-FH2 and TgFRM2-FH1-FH2 dramatically enhanced TgACTI polymerization in the absence of profilin, making them the sole protein factors known to initiate polymerization of this normally unstable actin. In addition, T. gondii formin domains were shown to both initiate polymerization and induce bundling of TgACTI filaments; however, they did not rely on TgPRF for these activities. In contrast, TgPRF sequestered TgACTI monomers, thus inhibiting polymerization even in the presence of formins. Collectively, these findings provide insight into the unusual control mechanisms of actin dynamics within the parasite.

  11. Crenactin forms actin-like double helical filaments regulated by arcadin-2

    PubMed Central

    Izoré, Thierry; Kureisaite-Ciziene, Danguole; McLaughlin, Stephen H; Löwe, Jan

    2016-01-01

    The similarity of eukaryotic actin to crenactin, a filament-forming protein from the crenarchaeon Pyrobaculum calidifontis supports the theory of a common origin of Crenarchaea and Eukaryotes. Monomeric structures of crenactin and actin are similar, although their filament architectures were suggested to be different. Here we report that crenactin forms bona fide double helical filaments that show exceptional similarity to eukaryotic F-actin. With cryo-electron microscopy and helical reconstruction we solved the structure of the crenactin filament to 3.8 Å resolution. When forming double filaments, the 'hydrophobic plug' loop in crenactin rearranges. Arcadin-2, also encoded by the arcade gene cluster, binds tightly with its C-terminus to the hydrophobic groove of crenactin. Binding is reminiscent of eukaryotic actin modulators such as cofilin and thymosin β4 and arcadin-2 is a depolymeriser of crenactin filaments. Our work further supports the theory of shared ancestry of Eukaryotes and Crenarchaea. DOI: http://dx.doi.org/10.7554/eLife.21600.001 PMID:27852434

  12. Aip1 Promotes Actin Filament Severing by Cofilin and Regulates Constriction of the Cytokinetic Contractile Ring*

    PubMed Central

    Chen, Qian; Courtemanche, Naomi; Pollard, Thomas D.

    2015-01-01

    Aip1 (actin interacting protein 1) is ubiquitous in eukaryotic organisms, where it cooperates with cofilin to disassemble actin filaments, but neither its mechanism of action nor its biological functions have been clear. We purified both fission yeast and human Aip1 and investigated their biochemical activities with or without cofilin. Both types of Aip1 bind actin filaments with micromolar affinities and weakly nucleate actin polymerization. Aip1 increases up to 12-fold the rate that high concentrations of yeast or human cofilin sever actin filaments, most likely by competing with cofilin for binding to the side of actin filaments, reducing the occupancy of the filaments by cofilin to a range favorable for severing. Aip1 does not cap the barbed ends of filaments severed by cofilin. Fission yeast lacking Aip1 are viable and assemble cytokinetic contractile rings normally, but rings in these Δaip1 cells accumulate 30% less myosin II. Further, these mutant cells initiate the ingression of cleavage furrows earlier than normal, shortening the stage of cytokinetic ring maturation by 50%. The Δaip1 mutation has negative genetic interactions with deletion mutations of both capping protein subunits and cofilin mutations with severing defects, but no genetic interaction with deletion of coronin. PMID:25451933

  13. HOW GALACTIC ENVIRONMENT REGULATES STAR FORMATION

    SciTech Connect

    Meidt, Sharon E.

    2016-02-10

    In a new simple model I reconcile two contradictory views on the factors that determine the rate at which molecular clouds form stars—internal structure versus external, environmental influences—providing a unified picture for the regulation of star formation in galaxies. In the presence of external pressure, the pressure gradient set up within a self-gravitating turbulent (isothermal) cloud leads to a non-uniform density distribution. Thus the local environment of a cloud influences its internal structure. In the simple equilibrium model, the fraction of gas at high density in the cloud interior is determined simply by the cloud surface density, which is itself inherited from the pressure in the immediate surroundings. This idea is tested using measurements of the properties of local clouds, which are found to show remarkable agreement with the simple equilibrium model. The model also naturally predicts the star formation relation observed on cloud scales and at the same time provides a mapping between this relation and the closer-to-linear molecular star formation relation measured on larger scales in galaxies. The key is that pressure regulates not only the molecular content of the ISM but also the cloud surface density. I provide a straightforward prescription for the pressure regulation of star formation that can be directly implemented in numerical models. Predictions for the dense gas fraction and star formation efficiency measured on large-scales within galaxies are also presented, establishing the basis for a new picture of star formation regulated by galactic environment.

  14. The Ustilago maydis Nit2 Homolog Regulates Nitrogen Utilization and Is Required for Efficient Induction of Filamentous Growth

    PubMed Central

    Horst, Robin J.; Zeh, Christine; Saur, Alexandra; Sonnewald, Sophia; Sonnewald, Uwe

    2012-01-01

    Nitrogen catabolite repression (NCR) is a regulatory strategy found in microorganisms that restricts the utilization of complex and unfavored nitrogen sources in the presence of favored nitrogen sources. In fungi, this concept has been best studied in yeasts and filamentous ascomycetes, where the GATA transcription factors Gln3p and Gat1p (in yeasts) and Nit2/AreA (in ascomycetes) constitute the main positive regulators of NCR. The reason why functional Nit2 homologs of some phytopathogenic fungi are required for full virulence in their hosts has remained elusive. We have identified the Nit2 homolog in the basidiomycetous phytopathogen Ustilago maydis and show that it is a major, but not the exclusive, positive regulator of nitrogen utilization. By transcriptome analysis of sporidia grown on artificial media devoid of favored nitrogen sources, we show that only a subset of nitrogen-responsive genes are regulated by Nit2, including the Gal4-like transcription factor Ton1 (a target of Nit2). Ustilagic acid biosynthesis is not under the control of Nit2, while nitrogen starvation-induced filamentous growth is largely dependent on functional Nit2. nit2 deletion mutants show the delayed initiation of filamentous growth on maize leaves and exhibit strongly compromised virulence, demonstrating that Nit2 is required to efficiently initiate the pathogenicity program of U. maydis. PMID:22247264

  15. Formation of dense structures induced by filament collisions. Correlation of density, kinematics, and magnetic field in the Pipe nebula

    NASA Astrophysics Data System (ADS)

    Frau, P.; Girart, J. M.; Alves, F. O.; Franco, G. A. P.; Onishi, T.; Román-Zúñiga, C. G.

    2015-02-01

    Context. The Pipe nebula is a molecular cloud that lacks star formation feedback and has a relatively simple morphology and velocity structure. This makes it an ideal target for testing cloud evolution through collisions. Aims: We aim at drawing a comprehensive picture of this relatively simple cloud to better understand the formation and evolution of molecular clouds on large scales. Methods: We use archival data to compare the optical polarization properties, the visual extinction, and the 13CO velocities and linewidths of the entire cloud in order to identify trends among the observables. Results: The Pipe nebula can be roughly divided into two filaments with different orientations and gas velocity ranges: E-W at 2-4 km s-1 and N-S at 6-7 km s-1. The two filaments overlap at the bowl, where the gas shows a velocity gradient spanning from 2 to 7 km s-1. Compared to the rest of the Pipe nebula, the bowl gas appears to be denser and exhibits larger linewidths. In addition, the polarization data at the bowl shows lower angular dispersion and higher polarization degree. Cores in the bowl tend to cluster in space and to follow the 13CO velocity gradient. In the stem, cores tend to cluster in regions with properties similar to those of the bowl. Conclusions: The velocity pattern points to a collision between the filaments in the bowl region. The magnetic field seems to be compressed and strengthened in the shocked region. The proportional increase in density and magnetic field strength by a factor similar to the Alfvénic Mach number suggests a continuous shock at low Alfvénic Mach number under the flux-freezing condition. Shocked regions seem to enhance the formation and clustering of dense cores. A movie associated to Fig. 2 is available at http://www.aanda.org

  16. Nucleoprotein filament formation is the structural basis for bacterial protein H-NS gene silencing

    NASA Astrophysics Data System (ADS)

    Lim, Ci Ji; Lee, Sin Yi; Kenney, Linda J.; Yan, Jie

    2012-07-01

    H-NS is an abundant nucleoid-associated protein in bacteria that globally silences genes, including horizontally-acquired genes related to pathogenesis. Although it has been shown that H-NS has multiple modes of DNA-binding, which mode is employed in gene silencing is still unclear. Here, we report that in H-NS mutants that are unable to silence genes, are unable to form a rigid H-NS nucleoprotein filament. These results indicate that the H-NS nucleoprotein filament is crucial for its gene silencing function, and serves as the fundamental structural basis for gene silencing by H-NS and likely other H-NS-like bacterial proteins.

  17. The identification of filaments on far-infrared and submillimiter images: Morphology, physical conditions and relation with star formation of filamentary structure

    SciTech Connect

    Schisano, E.; Carey, S.; Paladini, R.; Rygl, K. L. J.; Molinari, S.; Elia, D.; Pestalozzi, M.; Busquet, G.; Billot, N.; Noriega-Crespo, A.; Moore, T. J. T.; Plume, R.; Glover, S. C. O.; Vázquez-Semadeni, E.

    2014-08-10

    Observations of molecular clouds reveal a complex structure, with gas and dust often arranged in filamentary, rather than spherical geometries. The association of pre- and proto-stellar cores with the filaments suggests a direct link with the process of star formation. Any study of the properties of such filaments requires representative samples from different environments for an unbiased detection method. We developed such an approach using the Hessian matrix of a surface-brightness distribution to identify filaments and determine their physical and morphological properties. After testing the method on simulated, but realistic, filaments, we apply the algorithms to column-density maps computed from Herschel observations of the Galactic plane obtained by the Hi-GAL project. We identified ∼500 filaments, in the longitude range of l = 216.°5 to l = 225.°5, with lengths from ∼1 pc up to ∼30 pc and widths between 0.1 pc and 2.5 pc. Average column densities are between 10{sup 20} cm{sup –2} and 10{sup 22} cm{sup –2}. Filaments include the majority of dense material with N{sub H{sub 2}} > 6 × 10{sup 21} cm{sup –2}. We find that the pre- and proto-stellar compact sources already identified in the same region are mostly associated with filaments. However, surface densities in excess of the expected critical values for high-mass star formation are only found on the filaments, indicating that these structures are necessary to channel material into the clumps. Furthermore, we analyze the gravitational stability of filaments and discuss their relationship with star formation.

  18. The heterocyst differentiation transcriptional regulator HetR of the filamentous cyanobacterium Anabaena forms tetramers and can be regulated by phosphorylation.

    PubMed

    Valladares, Ana; Flores, Enrique; Herrero, Antonia

    2016-02-01

    Many filamentous cyanobacteria respond to the external cue of nitrogen scarcity by the differentiation of heterocysts, cells specialized in the fixation of atmospheric nitrogen in oxic environments. Heterocysts follow a spatial pattern along the filament of two heterocysts separated by ca. 10-15 vegetative cells performing oxygenic photosynthesis. HetR is a transcriptional regulator that directs heterocyst differentiation. In the model strain Anabaena sp. PCC 7120, the HetR protein was observed in various oligomeric forms in vivo, including a tetramer that peaked with maximal hetR expression during differentiation. Tetramers were not detected in a hetR point mutant incapable of differentiation, but were conspicuous in an over-differentiating strain lacking the PatS inhibitor. In differentiated filaments the HetR tetramer was restricted to heterocysts, being undetectable in vegetative cells. HetR co-purified with RNA polymerase from Anabaena mainly as a tetramer. In vitro, purified recombinant HetR was distributed between monomers, dimers, trimers and tetramers, and it was phosphorylated when incubated with (γ-(32)P)ATP. Phosphorylation and PatS hampered the accumulation of HetR tetramers and impaired HetR binding to DNA. In summary, tetrameric HetR appears to represent a functionally relevant form of HetR, whose abundance in the Anabaena filament could be negatively regulated by phosphorylation and by PatS.

  19. Hormonal and Local Regulation of Bone Formation.

    ERIC Educational Resources Information Center

    Canalis, Ernesto

    1985-01-01

    Reviews effects of hormones, systemic factors, and local regulators on bone formation. Identifies and explains the impact on bone growth of several hormones as well as the components of systemic and local systems. Concentrates on bone collagen and DNA synthesis. (Physicians may earn continuing education credit by completing an appended test). (ML)

  20. Hormonal and Local Regulation of Bone Formation.

    ERIC Educational Resources Information Center

    Canalis, Ernesto

    1985-01-01

    Reviews effects of hormones, systemic factors, and local regulators on bone formation. Identifies and explains the impact on bone growth of several hormones as well as the components of systemic and local systems. Concentrates on bone collagen and DNA synthesis. (Physicians may earn continuing education credit by completing an appended test). (ML)

  1. A network of filaments detected by Herschel in the Serpens core. A laboratory to test simulations of low-mass star formation

    NASA Astrophysics Data System (ADS)

    Roccatagliata, V.; Dale, J. E.; Ratzka, T.; Testi, L.; Burkert, A.; Koepferl, C.; Sicilia-Aguilar, A.; Eiroa, C.; Gaczkowski, B.

    2015-12-01

    Context. Filaments represent a key structure during the early stages of the star formation process. Simulations show that filamentary structures commonly formed before and during the formation of cores. Aims: The Serpens core is an ideal laboratory for testing the state of the art of simulations of turbulent giant molecular clouds. Methods: We used Herschel observations of the Serpens core to compute temperature and column density maps of the region. We selected the early stages of a recent simulation of star-formation, before stellar feedback was initiated, with similar total mass and physical size as the Serpens core. We also derived temperature and column density maps from the simulations. The observed distribution of column densities of the filaments was analyzed, first including and then masking the cores. The same analysis was performed on the simulations as well. Results: A radial network of filaments was detected in the Serpens core. The analyzed simulation shows a striking morphological resemblance to the observed structures. The column density distribution of simulated filaments without cores shows only a log-normal distribution, while the observed filaments show a power-law tail. The power-law tail becomes evident in the simulation if the focus is only the column density distribution of the cores. In contrast, the observed cores show a flat distribution. Conclusions: Even though the simulated and observed filaments are subjectively similar-looking, we find that they behave in very different ways. The simulated filaments are turbulence-dominated regions; the observed filaments are instead self-gravitating structures that will probably fragment into cores. Herschel is an ESA space observatory with science instruments provided by European-led Principal Investigator consortia and with important participation from NASA.Appendices are available in electronic form at http://www.aanda.org

  2. Multi-photon Imaging of Actin Filament Formation and Mitochondrial Energetics of Human ACBT Gliomas

    PubMed Central

    Hwang, Yu-Jer; Kolettis, Nomiki; Yang, Miso; Gillard, Elizabeth R.; Sanchez, Edgar; Sun, Chung-ho; Tromberg, Bruce J.; Krasieva, Tatiana B.; Lyubovitsky, Julia G.

    2011-01-01

    We studied the three-dimensional (3D) distribution of actin filaments and mitochondria in relation to ACBT glioblastoma cells migration. We embedded the cells in the spheroid form within collagen hydrogels and imaged them by in-situ multi-photon microscopy (MPM). The static 3D overlay of the distribution of actin filaments and mitochondria provided a greater understanding of cell-to-cell and cell-to-substrate interactions and morphology. While imaging mitochondria to obtain ratiometric redox index based on cellular fluorescence from reduced nicotinamide adenine dinucleotide (NADH) and oxidized flavin adenine dinucleotide (FAD) we observed differential sensitivity of the migrating ACBT glioblastoma cells to femtosecond laser irradiation employed in MPM. We imaged actin-GFP fluorescence in live ACBT glioma cells and for the first time observed dynamic modulation of the pools of actin during migration in 3D. The MPM imaging, which probes cells directly within the 3D cancer models, could potentially aid in working out a link between the functional performance of mitochondria, actin distribution and cancer invasiveness. PMID:21143483

  3. Candida albicans VPS1 contributes to protease secretion, filamentation, and biofilm formation.

    PubMed

    Bernardo, Stella M; Khalique, Zachary; Kot, John; Jones, Jason K; Lee, Samuel A

    2008-06-01

    To investigate the pre-vacuolar secretory pathway in Candida albicans, we cloned and analyzed the C. albicans homolog of the Saccharomyces cerevisiae vacuolar protein sorting gene VPS1. C. albicans VPS1 encodes a predicted 694-aa dynamin-like GTPase that is 73.3% similar to S. cerevisiae Vps1p. Plasmids bearing C. albicans VPS1 complemented the temperature-sensitive growth, abnormal class F vacuolar morphology, and carboxypeptidase missorting of a S. cerevisiae vps1 null mutant. To study VPS1 function in C. albicans, a conditional mutant strain (tetR-VPS1) was generated by deleting the first allele of VPS1 and placing the second allele under control of a tetracycline-regulatable promoter. With doxycycline, the tetR-VPS1 mutant was hyper-susceptible to sub-inhibitory concentrations of fluconazole, but not amphotericin B, 5-fluorocytosine, or non-specific osmotic stresses. The repressed tetR-VPS1 mutant was defective in filamentation and secreted less extracellular protease activity. Biofilm production and filamentation within the biofilm were markedly reduced. These results suggest that C. albicans VPS1 has a key role in several important virulence-related phenotypes.

  4. Regulators of Autophagosome Formation in Drosophila Muscles

    PubMed Central

    Zirin, Jonathan; Nieuwenhuis, Joppe; Samsonova, Anastasia; Tao, Rong; Perrimon, Norbert

    2015-01-01

    Given the diversity of autophagy targets and regulation, it is important to characterize autophagy in various cell types and conditions. We used a primary myocyte cell culture system to assay the role of putative autophagy regulators in the specific context of skeletal muscle. By treating the cultures with rapamycin (Rap) and chloroquine (CQ) we induced an autophagic response, fully suppressible by knockdown of core ATG genes. We screened D. melanogaster orthologs of a previously reported mammalian autophagy protein-protein interaction network, identifying several proteins required for autophagosome formation in muscle cells, including orthologs of the Rab regulators RabGap1 and Rab3Gap1. The screen also highlighted the critical roles of the proteasome and glycogen metabolism in regulating autophagy. Specifically, sustained proteasome inhibition inhibited autophagosome formation both in primary culture and larval skeletal muscle, even though autophagy normally acts to suppress ubiquitin aggregate formation in these tissues. In addition, analyses of glycogen metabolic genes in both primary cultured and larval muscles indicated that glycogen storage enhances the autophagic response to starvation, an important insight given the link between glycogen storage disorders, autophagy, and muscle function. PMID:25692684

  5. Regulation of flagellar motility during biofilm formation

    PubMed Central

    Guttenplan, Sarah B.; Kearns, Daniel B.

    2013-01-01

    Many bacteria swim in liquid or swarm over solid surfaces by synthesizing rotary flagella. The same bacteria that are motile also commonly form non-motile multicellular aggregates held together by an extracellular matrix called biofilms. Biofilms are an important part of the lifestyle of pathogenic bacteria and it is assumed that there is a motility-to-biofilm transition wherein the inhibition of motility promotes biofilm formation. The transition is largely inferred from regulatory mutants that reveal the opposite regulation of the two phenotypes. Here we review the regulation of motility during biofilm formation in Bacillus, Pseudomonas, Vibrio, and Escherichia, and we conclude that the motility-to-biofilm transition, if necessary, likely involves two steps. In the short term, flagella are functionally regulated to either inhibit rotation or modulate the basal flagellar reversal frequency. Over the long term, flagellar gene transcription is inhibited and in the absence of de novo synthesis, flagella are likely diluted to extinction through growth. Both short term and long term control is likely important to the motility-to-biofilm transition to stabilize aggregates and optimize resource investment. We emphasize the newly discovered classes of flagellar functional regulators and speculate that others await discovery in the context of biofilm formation. PMID:23480406

  6. The Interaction of Arp2/3 Complex with Actin: Nucleation, High Affinity Pointed End Capping, and Formation of Branching Networks of Filaments

    NASA Astrophysics Data System (ADS)

    Dyche Mullins, R.; Heuser, John A.; Pollard, Thomas D.

    1998-05-01

    The Arp2/3 complex is a stable assembly of seven protein subunits including two actin-related proteins (Arp2 and Arp3) and five novel proteins. Previous work showed that this complex binds to the sides of actin filaments and is concentrated at the leading edges of motile cells. Here, we show that Arp2/3 complex purified from Acanthamoeba caps the pointed ends of actin filaments with high affinity. Arp2/3 complex inhibits both monomer addition and dissociation at the pointed ends of actin filaments with apparent nanomolar affinity and increases the critical concentration for polymerization at the pointed end from 0.6 to 1.0 μ M. The high affinity of Arp2/3 complex for pointed ends and its abundance in amoebae suggest that in vivo all actin filament pointed ends are capped by Arp2/3 complex. Arp2/3 complex also nucleates formation of actin filaments that elongate only from their barbed ends. From kinetic analysis, the nucleation mechanism appears to involve stabilization of polymerization intermediates (probably actin dimers). In electron micrographs of quick-frozen, deep-etched samples, we see Arp2/3 bound to sides and pointed ends of actin filaments and examples of Arp2/3 complex attaching pointed ends of filaments to sides of other filaments. In these cases, the angle of attachment is a remarkably constant 70 ± 7 degrees. From these in vitro biochemical properties, we propose a model for how Arp2/3 complex controls the assembly of a branching network of actin filaments at the leading edge of motile cells.

  7. Atomistic mechanisms of copper filament formation and composition in Al2O3-based conductive bridge random access memory

    NASA Astrophysics Data System (ADS)

    Nail, C.; Blaise, P.; Molas, G.; Bernard, M.; Roule, A.; Toffoli, A.; Perniola, L.; Vallée, C.

    2017-07-01

    Conductive filament formation and composition in Oxide-based Conductive Bridge Random Access Memory (CBRAM) are investigated. To this end, Al2O3/Cu-based CBRAM is electrically characterized and studied. Current-voltage characteristics exhibit different forming behaviors depending on device polarization exposing the charged species involved during the forming process. In order to get more insights at the microscopic level, ion diffusion is investigated in depth by first-principles calculations. We study different point defects in Al2O3 which can come either from the post-process of the material itself or after top electrode deposition or during device operation. Since the role of Oxygen Vacancies (VO) and Copper (Cu) ions is core to the switching mechanism, ab initio calculations focus on their displacements. For different charge states in Al2O3, we extract the thermodynamic and activation energies of Cu, Te, Al, and O related point defects. The results reveal that Cu is not the only ion diffusing in the Al2O3-based CBRAM switching mechanism while Te ions appear unfavorable. A Cu/VO based hybrid filament model is proposed, and the impact of Aluminum Vacancies (VAl) on the forming process is demonstrated.

  8. Self-stabilized discharge filament in plane-parallel barrier discharge configuration: formation, breakdown mechanism, and memory effects

    NASA Astrophysics Data System (ADS)

    Tschiersch, R.; Nemschokmichal, S.; Bogaczyk, M.; Meichsner, J.

    2017-10-01

    Single self-stabilized discharge filaments were investigated in the plane-parallel electrode configuration. The barrier discharge was operated inside a gap of 3 mm shielded by glass plates to both electrodes, using helium-nitrogen mixtures and a square-wave feeding voltage at a frequency of 2 kHz. The combined application of electrical measurements, ICCD camera imaging, optical emission spectroscopy and surface charge diagnostics via the electro-optic Pockels effect allowed the correlation of the discharge development in the volume and on the dielectric surfaces. The formation criteria and existence regimes were found by systematic variation of the nitrogen admixture to helium, the total pressure and the feeding voltage amplitude. Single self-stabilized discharge filaments can be operated over a wide parameter range, foremost, by significant reduction of the voltage amplitude after the operation in the microdischarge regime. Here, the outstanding importance of the surface charge memory effect on the long-term stability was pointed out by the recalculated spatio-temporally resolved gap voltage. The optical emission revealed discharge characteristics that are partially reminiscent of both the glow-like barrier discharge and the microdischarge regime, such as a Townsend pre-phase, a fast cathode-directed ionization front during the breakdown and radially propagating surface discharges during the afterglow.

  9. Actin filaments regulate the adhesion between the plasma membrane and the cell wall of tobacco guard cells.

    PubMed

    Yu, Qin; Ren, Jing-Jing; Kong, Lan-Jing; Wang, Xiu-Ling

    2017-08-13

    During the opening and closing of stomata, guard cells undergo rapid and reversible changes in their volume and shape, which affects the adhesion of the plasma membrane (PM) to the cell wall (CW). The dynamics of actin filaments in guard cells are involved in stomatal movement by regulating structural changes and intracellular signaling. However, it is unclear whether actin dynamics regulate the adhesion of the PM to the CW. In this study, we investigated the relationship between actin dynamics and PM-CW adhesion by the hyperosmotic-induced plasmolysis of tobacco guard cells. We found that actin filaments in guard cells were depolymerized during mannitol-induced plasmolysis. The inhibition of actin dynamics by treatment with latrunculin B or jasplakinolide and the disruption of the adhesion between the PM and the CW by treatment with RGDS peptide (Arg-Gly-Asp-Ser) enhanced guard cell plasmolysis. However, treatment with latrunculin B alleviated the RGDS peptide-induced plasmolysis and endocytosis. Our results reveal that the actin depolymerization is involved in the regulation of the PW-CW adhesion during hyperosmotic-induced plasmolysis in tobacco guard cells.

  10. [Mechanisms and regulation of enzymatic hydrolysis of cellulose in filamentous fungi: classical cases and new models].

    PubMed

    Gutiérrez-Rojas, Ivonne; Moreno-Sarmiento, Nubia; Montoya, Dolly

    2015-01-01

    Cellulose is the most abundant renewable carbon source on earth. However, this polymer structure comprises a physical and chemical barrier for carbon access, which has limited its exploitation. In nature, only a few percentage of microorganisms may degrade this polymer by cellulase expression. Filamentous fungi are one of the most active and efficient groups among these microorganisms. This review describes similarities and differences between cellulase activity mechanisms and regulatory mechanisms controlling gene expression for 3 of the most studied cellulolytic filamentous fungi models: Trichoderma reesei, Aspergillus niger and Aspergillus nidulans, and the recently described model Neurospora crassa. Unlike gene expression mechanisms, it was found that enzymatic activity mechanisms are similar for all the studied models. Understanding the distinctive elements of each system is essential for the development of strategies for the improvement of cellulase production, either by providing the optimum environment (fermentation conditions) or increasing gene expression in these microorganisms by genetic engineering.

  11. The Arabidopsis microtubule-associated protein MAP65-3 supports infection by filamentous biotrophic pathogens by down-regulating salicylic acid-dependent defenses.

    PubMed

    Quentin, Michaël; Baurès, Isabelle; Hoefle, Caroline; Caillaud, Marie-Cécile; Allasia, Valérie; Panabières, Franck; Abad, Pierre; Hückelhoven, Ralph; Keller, Harald; Favery, Bruno

    2016-03-01

    The oomycete Hyaloperonospora arabidopsidis and the ascomycete Erysiphe cruciferarum are obligate biotrophic pathogens causing downy mildew and powdery mildew, respectively, on Arabidopsis. Upon infection, the filamentous pathogens induce the formation of intracellular bulbous structures called haustoria, which are required for the biotrophic lifestyle. We previously showed that the microtubule-associated protein AtMAP65-3 plays a critical role in organizing cytoskeleton microtubule arrays during mitosis and cytokinesis. This renders the protein essential for the development of giant cells, which are the feeding sites induced by root knot nematodes. Here, we show that AtMAP65-3 expression is also induced in leaves upon infection by the downy mildew oomycete and the powdery mildew fungus. Loss of AtMAP65-3 function in the map65-3 mutant dramatically reduced infection by both pathogens, predominantly at the stages of leaf penetration. Whole-transcriptome analysis showed an over-represented, constitutive activation of genes involved in salicylic acid (SA) biosynthesis, signaling, and defense execution in map65-3, whereas jasmonic acid (JA)-mediated signaling was down-regulated. Preventing SA synthesis and accumulation in map65-3 rescued plant susceptibility to pathogens, but not the developmental phenotype caused by cytoskeleton defaults. AtMAP65-3 thus has a dual role. It positively regulates cytokinesis, thus plant growth and development, and negatively interferes with plant defense against filamentous biotrophs. Our data suggest that downy mildew and powdery mildew stimulate AtMAP65-3 expression to down-regulate SA signaling for infection.

  12. Cooperation of Cdc42 small G protein-activating and actin filament-binding activities of frabin in microspike formation.

    PubMed

    Ikeda, W; Nakanishi, H; Tanaka, Y; Tachibana, K; Takai, Y

    2001-06-14

    Frabin is a GDP/GTP exchange protein for Cdc42 with actin filament (F-actin)-binding activity. Cdc42 is a small GTP-binding protein that forms filopodia-like microspikes in a variety of cells. Expression of frabin indeed forms microspikes through at least activation of Cdc42 in MDCK cells and fibroblasts such as COS7, L, and NIH3T3 cells. However, the role of the F-actin-binding activity of frabin in the microspike formation remains unknown. We have examined here this role of frabin by expressing various frabin mutants, which have lost Cdc42-activating or F-actin-binding activity, with or without a dominant active mutant of Cdc42 in MDCK and COS7 cells. We show here that for the microspike formation, either of the Cdc42-activating and F- actin-binding activities of frabin alone is not sufficient and both the activities are necessary and that both the activities play a cooperative role in the microspike formation. The present results, together with the earlier finding that Cdc42 reorganizes the actin cytoskeleton at least through the N-WASP-Arp2/3 complex, suggest that frabin directly and indirectly reorganizes the actin cytoskeleton through its F-actin-binding and Cdc42-activating activities, respectively, in a cooperative manner, eventually leading to microspike formation.

  13. The Respiratory Syncytial Virus Phosphoprotein, Matrix Protein, and Fusion Protein Carboxy-Terminal Domain Drive Efficient Filamentous Virus-Like Particle Formation.

    PubMed

    Meshram, Chetan D; Baviskar, Pradyumna S; Ognibene, Cherie M; Oomens, Antonius G P

    2016-12-01

    Virus-like particles (VLPs) are attractive as a vaccine concept. For human respiratory syncytial virus (hRSV), VLP assembly is poorly understood and appears inefficient. Hence, hRSV antigens are often incorporated into foreign VLP systems to generate anti-RSV vaccine candidates. To better understand the assembly, and ultimately to enable efficient production, of authentic hRSV VLPs, we examined the associated requirements and mechanisms. In a previous analysis in HEp-2 cells, the nucleoprotein (N), phosphoprotein (P), matrix protein (M), and fusion protein (F) were required for formation of filamentous VLPs, which, similar to those of wild-type virus, were associated with the cell surface. Using fluorescence and electron microscopy combined with immunogold labeling, we examined the surfaces of transfected HEp-2 cells and further dissected the process of filamentous VLP formation. Our results show that N is not required. Coexpression of P plus M plus F, but not P plus M, M plus F, or P plus F, induced both viral protein coalescence and formation of filamentous VLPs that resembled wild-type virions. Despite suboptimal coalescence in the absence of P, the M and F proteins, when coexpressed, formed cell surface-associated filaments with abnormal morphology, appearing longer and thinner than wild-type virions. For F, only the carboxy terminus (Fstem) was required, and addition of foreign protein sequences to Fstem allowed incorporation into VLPs. Together, the data show that P, M, and the F carboxy terminus are sufficient for robust viral protein coalescence and filamentous VLP formation and suggest that M-F interaction drives viral filament formation, with P acting as a type of cofactor facilitating the process and exerting control over particle morphology.

  14. Microtubule-dependent transport of vimentin filament precursors is regulated by actin and by the concerted action of Rho- and p21-activated kinases.

    PubMed

    Robert, Amélie; Herrmann, Harald; Davidson, Michael W; Gelfand, Vladimir I

    2014-07-01

    Intermediate filaments (IFs) form a dense and dynamic network that is functionally associated with microtubules and actin filaments. We used the GFP-tagged vimentin mutant Y117L to study vimentin-cytoskeletal interactions and transport of vimentin filament precursors. This mutant preserves vimentin interaction with other components of the cytoskeleton, but its assembly is blocked at the unit-length filament (ULF) stage. ULFs are easy to track, and they allow a reliable and quantifiable analysis of movement. Our results show that in cultured human vimentin-negative SW13 cells, 2% of vimentin-ULFs move along microtubules bidirectionally, while the majority are stationary and tightly associated with actin filaments. Rapid motor-dependent transport of ULFs along microtubules is enhanced ≥ 5-fold by depolymerization of actin cytoskeleton with latrunculin B. The microtubule-dependent transport of vimentin ULFs is further regulated by Rho-kinase (ROCK) and p21-activated kinase (PAK): ROCK inhibits ULF transport, while PAK stimulates it. Both kinases act on microtubule transport independently of their effects on actin cytoskeleton. Our study demonstrates the importance of the actin cytoskeleton to restrict IF transport and reveals a new role for PAK and ROCK in the regulation of IF precursor transport.-Robert, A., Herrmann, H., Davidson, M. W., and Gelfand, V. I. Microtubule-dependent transport of vimentin filament precursors is regulated by actin and by the concerted action of Rho- and p21-activated kinases.

  15. Global Simulations of Differentially Rotating Magnetized Disks: Formation of Low-beta Filaments and Structured Coronae.

    PubMed

    Machida; Hayashi; Matsumoto

    2000-03-20

    We present the results of three-dimensional global magnetohydrodynamic simulations of the Parker-shearing instability in a differentially rotating torus initially threaded by toroidal magnetic fields. An equilibrium model of a magnetized torus is adopted as an initial condition. When beta0=Pgas&solm0;Pmag approximately 1 at the initial state, magnetic flux buoyantly escapes from the disk and creates looplike structures similar to those in the solar corona. Inside the torus, the growth of nonaxisymmetric magnetorotational (or Balbus & Hawley) instability generates magnetic turbulence. Magnetic field lines are tangled on a small scale, but on a large scale they show low azimuthal wavenumber spiral structure. After several rotation periods, the system oscillates around a state with beta approximately 5. We found that magnetic pressure-dominated (beta<1) filaments are created in the torus. The volume filling factor of the region in which beta

  16. The role of cyclase-associated protein in regulating actin filament dynamics - more than a monomer-sequestration factor.

    PubMed

    Ono, Shoichiro

    2013-08-01

    Dynamic reorganization of the actin cytoskeleton is fundamental to a number of cell biological events. A variety of actin-regulatory proteins modulate polymerization and depolymerization of actin and contribute to actin cytoskeletal reorganization. Cyclase-associated protein (CAP) is a conserved actin-monomer-binding protein that has been studied for over 20 years. Early studies have shown that CAP sequesters actin monomers; recent studies, however, have revealed more active roles of CAP in actin filament dynamics. CAP enhances the recharging of actin monomers with ATP antagonistically to ADF/cofilin, and also promotes the severing of actin filaments in cooperation with ADF/cofilin. Self-oligomerization and binding to other proteins regulate activities and localization of CAP. CAP has crucial roles in cell signaling, development, vesicle trafficking, cell migration and muscle sarcomere assembly. This Commentary discusses the recent advances in our understanding of the functions of CAP and its implications as an important regulator of actin cytoskeletal dynamics, which are involved in various cellular activities.

  17. Force maintenance and myosin filament assembly regulated by Rho-kinase in airway smooth muscle.

    PubMed

    Lan, Bo; Deng, Linhong; Donovan, Graham M; Chin, Leslie Y M; Syyong, Harley T; Wang, Lu; Zhang, Jenny; Pascoe, Christopher D; Norris, Brandon A; Liu, Jeffrey C-Y; Swyngedouw, Nicholas E; Banaem, Saleha M; Paré, Peter D; Seow, Chun Y

    2015-01-01

    Smooth muscle contraction can be divided into two phases: the initial contraction determines the amount of developed force and the second phase determines how well the force is maintained. The initial phase is primarily due to activation of actomyosin interaction and is relatively well understood, whereas the second phase remains poorly understood. Force maintenance in the sustained phase can be disrupted by strains applied to the muscle; the strain causes actomyosin cross-bridges to detach and also the cytoskeletal structure to disassemble in a process known as fluidization, for which the underlying mechanism is largely unknown. In the present study we investigated the ability of airway smooth muscle to maintain force after the initial phase of contraction. Specifically, we examined the roles of Rho-kinase and protein kinase C (PKC) in force maintenance. We found that for the same degree of initial force inhibition, Rho-kinase substantially reduced the muscle's ability to sustain force under static conditions, whereas inhibition of PKC had a minimal effect on sustaining force. Under oscillatory strain, Rho-kinase inhibition caused further decline in force, but again, PKC inhibition had a minimal effect. We also found that Rho-kinase inhibition led to a decrease in the myosin filament mass in the muscle cells, suggesting that one of the functions of Rho-kinase is to stabilize myosin filaments. The results also suggest that dissolution of myosin filaments may be one of the mechanisms underlying the phenomenon of fluidization. These findings can shed light on the mechanism underlying deep inspiration induced bronchodilation.

  18. CAP1, an adenylate cyclase-associated protein gene, regulates bud-hypha transitions, filamentous growth, and cyclic AMP levels and is required for virulence of Candida albicans.

    PubMed

    Bahn, Y S; Sundstrom, P

    2001-05-01

    In response to a wide variety of environmental stimuli, the opportunistic fungal pathogen Candida albicans exits the budding cycle, producing germ tubes and hyphae concomitant with expression of virulence genes, such as that encoding hyphal wall protein 1 (HWP1). Biochemical studies implicate cyclic AMP (cAMP) increases in promoting bud-hypha transitions, but genetic evidence relating genes that control cAMP levels to bud-hypha transitions has not been reported. Adenylate cyclase-associated proteins (CAPs) of nonpathogenic fungi interact with Ras and adenylate cyclase to increase cAMP levels under specific environmental conditions. To initiate studies on the relationship between cAMP signaling and bud-hypha transitions in C. albicans, we identified, cloned, characterized, and disrupted the C. albicans CAP1 gene. C. albicans strains with inactivated CAP1 budded in conditions that led to germ tube formation in isogenic strains with CAP1. The addition of 10 mM cAMP and dibutyryl cAMP promoted bud-hypha transitions and filamentous growth in the cap1/cap1 mutant in liquid and solid media, respectively, showing clearly that cAMP promotes hypha formation in C. albicans. Increases in cytoplasmic cAMP preceding germ tube emergence in strains having CAP1 were markedly diminished in the budding cap1/cap1 mutant. C. albicans strains with deletions of both alleles of CAP1 were avirulent in a mouse model of systemic candidiasis. The avirulence of a germ tube-deficient cap1/cap1 mutant coupled with the role of Cap1 in regulating cAMP levels shows that the Cap1-mediated cAMP signaling pathway is required for bud-hypha transitions, filamentous growth, and the pathogenesis of candidiasis.

  19. CAP1, an Adenylate Cyclase-Associated Protein Gene, Regulates Bud-Hypha Transitions, Filamentous Growth, and Cyclic AMP Levels and Is Required for Virulence of Candida albicans

    PubMed Central

    Bahn, Yong-Sun; Sundstrom, Paula

    2001-01-01

    In response to a wide variety of environmental stimuli, the opportunistic fungal pathogen Candida albicans exits the budding cycle, producing germ tubes and hyphae concomitant with expression of virulence genes, such as that encoding hyphal wall protein 1 (HWP1). Biochemical studies implicate cyclic AMP (cAMP) increases in promoting bud-hypha transitions, but genetic evidence relating genes that control cAMP levels to bud-hypha transitions has not been reported. Adenylate cyclase-associated proteins (CAPs) of nonpathogenic fungi interact with Ras and adenylate cyclase to increase cAMP levels under specific environmental conditions. To initiate studies on the relationship between cAMP signaling and bud-hypha transitions in C. albicans, we identified, cloned, characterized, and disrupted the C. albicans CAP1 gene. C. albicans strains with inactivated CAP1 budded in conditions that led to germ tube formation in isogenic strains with CAP1. The addition of 10 mM cAMP and dibutyryl cAMP promoted bud-hypha transitions and filamentous growth in the cap1/cap1 mutant in liquid and solid media, respectively, showing clearly that cAMP promotes hypha formation in C. albicans. Increases in cytoplasmic cAMP preceding germ tube emergence in strains having CAP1 were markedly diminished in the budding cap1/cap1 mutant. C. albicans strains with deletions of both alleles of CAP1 were avirulent in a mouse model of systemic candidiasis. The avirulence of a germ tube-deficient cap1/cap1 mutant coupled with the role of Cap1 in regulating cAMP levels shows that the Cap1-mediated cAMP signaling pathway is required for bud-hypha transitions, filamentous growth, and the pathogenesis of candidiasis. PMID:11325951

  20. Covalent modification of DNA regulates memory formation.

    PubMed

    Miller, Courtney A; Sweatt, J David

    2007-03-15

    DNA methylation is a covalent chemical modification of DNA catalyzed by DNA methyltransferases (DNMTs). DNA methylation is associated with transcriptional silencing and has been studied extensively as a lifelong molecular information storage mechanism put in place during development. Here we report that DNMT gene expression is upregulated in the adult rat hippocampus following contextual fear conditioning and that DNMT inhibition blocks memory formation. In addition, fear conditioning is associated with rapid methylation and transcriptional silencing of the memory suppressor gene PP1 and demethylation and transcriptional activation of the synaptic plasticity gene reelin, indicating both methyltransferase and demethylase activity during consolidation. DNMT inhibition prevents the PP1 methylation increase, resulting in aberrant transcription of the gene during the memory-consolidation period. These results demonstrate that DNA methylation is dynamically regulated in the adult nervous system and that this cellular mechanism is a crucial step in memory formation.

  1. The RNAi machinery regulates growth and development in the filamentous fungus Trichoderma atroviride.

    PubMed

    Carreras-Villaseñor, Nohemi; Esquivel-Naranjo, Edgardo U; Villalobos-Escobedo, J Manuel; Abreu-Goodger, Cei; Herrera-Estrella, Alfredo

    2013-07-01

    The RNAi machinery is generally involved in genome protection in filamentous fungi; however, the physiological role of RNAi has been poorly studied in fungal models. Here, we report that in the filamentous fungus Trichoderma atroviride, the products of the dcr2 and rdr3 genes control reproductive development, because mutations in these genes affect conidiation. In addition, Dcr1 together with Dcr2 control vegetative growth since Δdcr1, Δdcr2 and Δdcr1Δdcr2 present morphological alterations. Whole-genome transcriptional analysis of WT, Δdcr1, Δdcr2 and Δdcr1Δdcr2 show that each Dicer controls different biological processes, such as development or metabolism, which could explain the lack of conidiation in the mutants. Finally, we observed sRNAs that are differentially expressed in the WT and Δdcr2. The expression of some of these sRNAs correlates with the expression of differential transcripts, suggesting that these mRNAs may contain the corresponding targets. Together these data show that in T. atroviride, the RNAi machinery plays a central role in endogenous processes such as development and fitness, beyond controlling genome protection against invasive nucleic acids as reported for other fungi.

  2. The role of disulfide bond formation in the structural transition observed in the intermediate filaments of developing hair.

    PubMed

    Bruce Fraser, R D; Parry, David A D

    2012-10-01

    Hair keratin is a composite structure in which intermediate filaments (IF) are embedded in a protein matrix. During the early stages of development in the hair follicle the redox potential is such that the cysteine residues in the IF are maintained in a reduced form. However, at a late stage of development the redox potential changes to produce an oxidizing environment and the IF undergo a structural transition involving both molecular slippage and radial compaction. In our earlier study the changes in the molecular parameters were estimated from knowledge of the sites of artificially induced crosslinks, and it was noted that the changes in these parameters realigned many of the cysteine residues to positions more favorable to disulfide bond formation. As the energy involved in the formation of disulfide bonds is much greater than that of hydrogen bonds or van der Waals interactions the structural transition is likely to be dominated by the requirement that the bonded cysteine residues occur at closely equivalent axial positions. This criterion was used in the present study to obtain more precise values for the molecular parameters in the oxidized fiber than has hitherto been possible. A comparison of the sequences of hair keratins and epidermal keratins suggests that the slippage observed in trichocyte IF during keratinization does not occur in epidermal IF. Copyright © 2012 Elsevier Inc. All rights reserved.

  3. Branching of keratin intermediate filaments.

    PubMed

    Nafeey, Soufi; Martin, Ines; Felder, Tatiana; Walther, Paul; Felder, Edward

    2016-06-01

    Keratin intermediate filaments (IFs) are crucial to maintain mechanical stability in epithelial cells. Since little is known about the network architecture that provides this stiffness and especially about branching properties of filaments, we addressed this question with different electron microscopic (EM) methods. Using EM tomography of high pressure frozen keratinocytes, we investigated the course of several filaments in a branching of a filament bundle. Moreover we found several putative bifurcations in individual filaments. To verify our observation we also visualized the keratin network in detergent extracted keratinocytes with scanning EM. Here bifurcations of individual filaments could unambiguously be identified additionally to bundle branchings. Interestingly, identical filament bifurcations were also found in purified keratin 8/18 filaments expressed in Escherichia coli which were reassembled in vitro. This excludes that an accessory protein contributes to the branch formation. Measurements of the filament cross sectional areas showed various ratios between the three bifurcation arms. This demonstrates that intermediate filament furcation is very different from actin furcation where an entire new filament is attached to an existing filament. Instead, the architecture of intermediate filament bifurcations is less predetermined and hence consistent with the general concept of IF formation. Copyright © 2016 Elsevier Inc. All rights reserved.

  4. FtsZ filament capping by MciZ, a developmental regulator of bacterial division

    PubMed Central

    Bisson-Filho, Alexandre W.; Discola, Karen F.; Castellen, Patrícia; Blasios, Valdir; Martins, Alexandre; Sforça, Maurício L.; Garcia, Wanius; Zeri, Ana Carolina M.; Erickson, Harold P.; Dessen, Andréa; Gueiros-Filho, Frederico J.

    2015-01-01

    Cytoskeletal structures are dynamically remodeled with the aid of regulatory proteins. FtsZ (filamentation temperature-sensitive Z) is the bacterial homolog of tubulin that polymerizes into rings localized to cell-division sites, and the constriction of these rings drives cytokinesis. Here we investigate the mechanism by which the Bacillus subtilis cell-division inhibitor, MciZ (mother cell inhibitor of FtsZ), blocks assembly of FtsZ. The X-ray crystal structure reveals that MciZ binds to the C-terminal polymerization interface of FtsZ, the equivalent of the minus end of tubulin. Using in vivo and in vitro assays and microscopy, we show that MciZ, at substoichiometric levels to FtsZ, causes shortening of protofilaments and blocks the assembly of higher-order FtsZ structures. The findings demonstrate an unanticipated capping-based regulatory mechanism for FtsZ. PMID:25848052

  5. FtsZ filament capping by MciZ, a developmental regulator of bacterial division.

    PubMed

    Bisson-Filho, Alexandre W; Discola, Karen F; Castellen, Patrícia; Blasios, Valdir; Martins, Alexandre; Sforça, Maurício L; Garcia, Wanius; Zeri, Ana Carolina M; Erickson, Harold P; Dessen, Andréa; Gueiros-Filho, Frederico J

    2015-04-28

    Cytoskeletal structures are dynamically remodeled with the aid of regulatory proteins. FtsZ (filamentation temperature-sensitive Z) is the bacterial homolog of tubulin that polymerizes into rings localized to cell-division sites, and the constriction of these rings drives cytokinesis. Here we investigate the mechanism by which the Bacillus subtilis cell-division inhibitor, MciZ (mother cell inhibitor of FtsZ), blocks assembly of FtsZ. The X-ray crystal structure reveals that MciZ binds to the C-terminal polymerization interface of FtsZ, the equivalent of the minus end of tubulin. Using in vivo and in vitro assays and microscopy, we show that MciZ, at substoichiometric levels to FtsZ, causes shortening of protofilaments and blocks the assembly of higher-order FtsZ structures. The findings demonstrate an unanticipated capping-based regulatory mechanism for FtsZ.

  6. Epigenetic regulation of memory formation and maintenance.

    PubMed

    Zovkic, Iva B; Guzman-Karlsson, Mikael C; Sweatt, J David

    2013-01-15

    Understanding the cellular and molecular mechanisms underlying the formation and maintenance of memories is a central goal of the neuroscience community. It is well regarded that an organism's ability to lastingly adapt its behavior in response to a transient environmental stimulus relies on the central nervous system's capability for structural and functional plasticity. This plasticity is dependent on a well-regulated program of neurotransmitter release, post-synaptic receptor activation, intracellular signaling cascades, gene transcription, and subsequent protein synthesis. In the last decade, epigenetic markers like DNA methylation and post-translational modifications of histone tails have emerged as important regulators of the memory process. Their ability to regulate gene transcription dynamically in response to neuronal activation supports the consolidation of long-term memory. Furthermore, the persistent and self-propagating nature of these mechanisms, particularly DNA methylation, suggests a molecular mechanism for memory maintenance. In this review, we will examine the evidence that supports a role of epigenetic mechanisms in learning and memory. In doing so, we hope to emphasize (1) the widespread involvement of these mechanisms across different behavioral paradigms and distinct brain regions, (2) the temporal and genetic specificity of these mechanisms in response to upstream signaling cascades, and (3) the functional outcome these mechanisms may have on structural and functional plasticity. Finally, we consider the future directions of neuroepigenetic research as it relates to neuronal storage of information.

  7. Epigenetic regulation of memory formation and maintenance

    PubMed Central

    Zovkic, Iva B.; Guzman-Karlsson, Mikael C.; Sweatt, J. David

    2013-01-01

    Understanding the cellular and molecular mechanisms underlying the formation and maintenance of memories is a central goal of the neuroscience community. It is well regarded that an organism's ability to lastingly adapt its behavior in response to a transient environmental stimulus relies on the central nervous system's capability for structural and functional plasticity. This plasticity is dependent on a well-regulated program of neurotransmitter release, post-synaptic receptor activation, intracellular signaling cascades, gene transcription, and subsequent protein synthesis. In the last decade, epigenetic markers like DNA methylation and post-translational modifications of histone tails have emerged as important regulators of the memory process. Their ability to regulate gene transcription dynamically in response to neuronal activation supports the consolidation of long-term memory. Furthermore, the persistent and self-propagating nature of these mechanisms, particularly DNA methylation, suggests a molecular mechanism for memory maintenance. In this review, we will examine the evidence that supports a role of epigenetic mechanisms in learning and memory. In doing so, we hope to emphasize (1) the widespread involvement of these mechanisms across different behavioral paradigms and distinct brain regions, (2) the temporal and genetic specificity of these mechanisms in response to upstream signaling cascades, and (3) the functional outcome these mechanisms may have on structural and functional plasticity. Finally, we consider the future directions of neuroepigenetic research as it relates to neuronal storage of information. PMID:23322554

  8. Studying Pellet Formation of a Filamentous Fungus Rhizopus oryzae to Enhance Organic Acid Production

    NASA Astrophysics Data System (ADS)

    Liao, Wei; Liu, Yan; Chen, Shulin

    Using pelletized fungal biomass can effectively improve the fermentation performance for most of fugal strains. This article studied the effects of inoculum and medium compositions such as potato dextrose broth (PDB) as carbon source, soybean peptone, calcium carbonate, and metal ions on pellet formation of Rhizopus oryzae. It has been found that metal ions had significantly negative effects on pellet formation whereas soybean peptone had positive effects. In addition PDB and calcium carbonate were beneficial to R. oryzae for growing small smooth pellets during the culture. The study also demonstrated that an inoculum size of less than 1.5×109 spores/L had no significant influence on pellet formation. Thus, a new approach to form pellets has been developed using only PDB, soybean peptone, and calcium carbonate. Meanwhile, palletized fungal fermentation significantly enhanced organic acid production. Lactic acid concentration reached 65.0 g/L in 30 h using pelletized R. oryzae NRRL 395, and fumeric acid concentration reached 31.0 g/L in 96 h using pelletized R. oryzae ATCC 20344.

  9. In vivo formation steps of the hard alpha-keratin intermediate filament along a hair follicle: evidence for structural polymorphism.

    PubMed

    Rafik, Mériem Er; Briki, Fatma; Burghammer, Manfred; Doucet, Jean

    2006-04-01

    Several aspects of the intermediate filaments' molecular architecture remain mysterious despite decades of study. The growth process and the final architecture may depend on the physical, chemical, and biochemical environment. Aiming at clarifying this issue, we have revisited the structure of the human hair follicle by means of X-ray microdiffraction. We conclude that the histology-based growth zones along the follicle are correlated to the fine architecture of the filaments deduced from X-ray microdiffraction. Our analysis reveals the existence of two major polymorph intermediate filament architectures. Just above the bulb, the filaments are characterized by a diameter of 100 Angstroms and a low-density core. The following zone upwards is characterized by the lateral aggregation of the filaments into a compact network of filaments, by a contraction of their diameter (to 75 Angstroms) and by the setting up of a long-range longitudinal ordering. In the upper zone, the small structural change associated with the tissue hardening likely concerns the terminal domains. The architecture of the intermediate filament in the upper zones could be specific to hard alpha-keratin whilst the other architecture found in the lower zone could be representative for intermediate filaments in a different environment.

  10. Formation and Initiation of Erupting Flux Rope and Embedded Filament Driven by Photospheric Converging Motion

    NASA Astrophysics Data System (ADS)

    Zhao, Xiaozhou; Xia, Chun; Keppens, Rony; Gan, Weiqun

    2017-06-01

    In this paper, we study how a flux rope (FR) is formed and evolves into the corresponding structure of a coronal mass ejection (CME) numerically driven by photospheric converging motion. A two-and-a-half-dimensional magnetohydrodynamics simulation is conducted in a chromosphere-transition-corona setup. The initial arcade-like linear force-free magnetic field is driven by an imposed slow motion converging toward the magnetic inversion line at the bottom boundary. The convergence brings opposite-polarity magnetic flux to the polarity inversion, giving rise to the formation of an FR by magnetic reconnection and eventually to the eruption of a CME. During the FR formation, an embedded prominence gets formed by the levitation of chromospheric material. We confirm that the converging flow is a potential mechanism for the formation of FRs and a possible triggering mechanism for CMEs. We investigate the thermal, dynamical, and magnetic properties of the FR and its embedded prominence by tracking their thermal evolution, analyzing their force balance, and measuring their kinematic quantities. The phase transition from the initiation phase to the acceleration phase of the kinematic evolution of the FR was observed in our simulation. The FR undergoes a series of quasi-static equilibrium states in the initiation phase; while in the acceleration phase the FR is driven by Lorentz force and the impulsive acceleration occurs. The underlying physical reason for the phase transition is the change of the reconnection mechanism from the Sweet-Parker to the unsteady bursty regime of reconnection in the evolving current sheet underneath the FR.

  11. Feedback Regulated Star Formation in Cool Core Clusters of Galaxies

    NASA Astrophysics Data System (ADS)

    Tremblay, Grant Russell

    2011-07-01

    The classical "cooling flow" model historically associated with "cool core" clusters of galaxies fails in the absence of an external, non-gravitational heating mechanism needed to offset catastrophic radiative losses of the X-ray bright intracluster medium (ICM). Numerous proposed solutions exist, including feedback from active galactic nuclei (AGN), which may elegantly calibrate fundamental relationships such as the coupled co-evolution of black holes and the stellar component of their host galaxies. AGN feedback cannot completely offset cooling at all times, however, as the brightest cluster galaxies (BCGs) in cool core clusters harbor extensive warm (˜104 K) and cold (10 < T < 104 K) gas reservoirs whose physical properties are regulated by ongoing star formation and an unknown, non-stellar heating mechanism. We present a doctoral thesis broadly related to these issues, particularly as they pertain to cooling flows, the triggering of AGN activity, and the associated energetic feedback that may play a critical role in heating the ambient environment on tens to hundreds of kiloparsec scales. We begin with a summary of the relevant background material, and in Chapter 2 we present a multiwavelength study of effervescent AGN heating in the cool core cluster Abell 2597. Previously unpublished Chandra X-ray data show the central regions of the hot intracluster medium (ICM) to be highly anisotropic on the scale of the BCG, permeated by a network of kpc-scale X-ray cavities, the largest of which is cospatial in projection with extended 330 MHz radio emission. We present spectral maps of projected, modeled gas properties fit to the X-ray data. The X-ray temperature map reveals two discrete, "hard-edged'' structures, including a ˜15 kpc "cold filament'' and an arc of hot gas which in projection borders the inner edge of the large X-ray cavity. We interpret the latter in the context of the effervescent AGN heating model, in which cavity enthalpy is thermalized as the

  12. Candida albicans White and Opaque Cells Undergo Distinct Programs of Filamentous Growth

    PubMed Central

    Si, Haoyu; Hernday, Aaron D.; Hirakawa, Matthew P.; Johnson, Alexander D.; Bennett, Richard J.

    2013-01-01

    The ability to switch between yeast and filamentous forms is central to Candida albicans biology. The yeast-hyphal transition is implicated in adherence, tissue invasion, biofilm formation, phagocyte escape, and pathogenesis. A second form of morphological plasticity in C. albicans involves epigenetic switching between white and opaque forms, and these two states exhibit marked differences in their ability to undergo filamentation. In particular, filamentous growth in white cells occurs in response to a number of environmental conditions, including serum, high temperature, neutral pH, and nutrient starvation, whereas none of these stimuli induce opaque filamentation. Significantly, however, we demonstrate that opaque cells can undergo efficient filamentation but do so in response to distinct environmental cues from those that elicit filamentous growth in white cells. Growth of opaque cells in several environments, including low phosphate medium and sorbitol medium, induced extensive filamentous growth, while white cells did not form filaments under these conditions. Furthermore, while white cell filamentation is often enhanced at elevated temperatures such as 37°C, opaque cell filamentation was optimal at 25°C and was inhibited by higher temperatures. Genetic dissection of the opaque filamentation pathway revealed overlapping regulation with the filamentous program in white cells, including key roles for the transcription factors EFG1, UME6, NRG1 and RFG1. Gene expression profiles of filamentous white and opaque cells were also compared and revealed only limited overlap between these programs, although UME6 was induced in both white and opaque cells consistent with its role as master regulator of filamentation. Taken together, these studies establish that a program of filamentation exists in opaque cells. Furthermore, this program regulates a distinct set of genes and is under different environmental controls from those operating in white cells. PMID:23505370

  13. Candida albicans white and opaque cells undergo distinct programs of filamentous growth.

    PubMed

    Si, Haoyu; Hernday, Aaron D; Hirakawa, Matthew P; Johnson, Alexander D; Bennett, Richard J

    2013-03-01

    The ability to switch between yeast and filamentous forms is central to Candida albicans biology. The yeast-hyphal transition is implicated in adherence, tissue invasion, biofilm formation, phagocyte escape, and pathogenesis. A second form of morphological plasticity in C. albicans involves epigenetic switching between white and opaque forms, and these two states exhibit marked differences in their ability to undergo filamentation. In particular, filamentous growth in white cells occurs in response to a number of environmental conditions, including serum, high temperature, neutral pH, and nutrient starvation, whereas none of these stimuli induce opaque filamentation. Significantly, however, we demonstrate that opaque cells can undergo efficient filamentation but do so in response to distinct environmental cues from those that elicit filamentous growth in white cells. Growth of opaque cells in several environments, including low phosphate medium and sorbitol medium, induced extensive filamentous growth, while white cells did not form filaments under these conditions. Furthermore, while white cell filamentation is often enhanced at elevated temperatures such as 37°C, opaque cell filamentation was optimal at 25°C and was inhibited by higher temperatures. Genetic dissection of the opaque filamentation pathway revealed overlapping regulation with the filamentous program in white cells, including key roles for the transcription factors EFG1, UME6, NRG1 and RFG1. Gene expression profiles of filamentous white and opaque cells were also compared and revealed only limited overlap between these programs, although UME6 was induced in both white and opaque cells consistent with its role as master regulator of filamentation. Taken together, these studies establish that a program of filamentation exists in opaque cells. Furthermore, this program regulates a distinct set of genes and is under different environmental controls from those operating in white cells.

  14. The role of filamentous hemagglutinin adhesin in adherence and biofilm formation in Acinetobacter baumannii ATCC19606(T).

    PubMed

    Darvish Alipour Astaneh, Shakiba; Rasooli, Iraj; Mousavi Gargari, Seyed Latif

    2014-09-01

    Filamentous hemagglutinin adhesins (FHA) are key factors for bacterial attachment and subsequent cell accumulation on substrates. Here an FHA-like Outer membrane (OM) adhesin of Acinetobacter baumannii ATCC19606(T) was displayed on Escherichia coli. The candidate autotransporter (AT) genes were identified in A. baumannii ATCC19606(T) genome. The exoprotein (FhaB1) and transporter (FhaC1) were produced independently within the same cell (FhaB1C1). The fhaC1 was mutated. In vitro adherence to epithelial cells of the recombinant FhaB1C1 and the mutant strains were compared with A. baumanni ATCC19606(T). A bivalent chimeric protein (K) composed of immunologically important portions of fhaB1 (B) and fhaC1 (C) was constructed. The mice vaccinated with chimeric protein were challenged with A. baumannii ATCC19606(T) and FhaB1C1 producing recombinant E. coli. Mutations in the fhaC1 resulted in the absence of FhaB1 in the OM. Expression of FhaB1C1 enhanced the adherence of recombinant bacteria to A546 bronchial cell line. The results revealed association of FhaB1 with bacterial adhesion and biofilm formation. Immunization with a combination of recombinant B and K proteins proved protective against A. baumanni ATCC19606(T). The findings may be applied in active and passive immunization strategies against A. baumannii.

  15. Regulation of hard α-keratin mechanics via control of intermediate filament hydration: matrix squeeze revisited.

    PubMed

    Greenberg, Daniel A; Fudge, Douglas S

    2013-01-07

    Mammalian hard α-keratins are fibre-reinforced biomaterials that consist of 10 nm intermediate filaments (IFs) embedded in an elastomeric protein matrix. Recent work suggests that the mechanical properties of IFs are highly sensitive to hydration, whereas hard α-keratins such as wool, hair and nail are relatively hydration insensitive. This raises the question of how mammalian keratins remain stiff in water. The matrix squeeze hypothesis states that the IFs in hard α-keratins are stiffened during an air-drying step during keratinization, and subsequently locked into a dehydrated state via the oxidation and cross-linking of the keratin matrix around them. The result is that even when hard α-keratins are immersed in water, their constituent IFs remain essentially 'dry' and therefore stiff. This hypothesis makes several predictions about the effects of matrix abundance and function on hard α-keratin mechanics and swelling behaviour. Specifically, it predicts that high matrix keratins in water will swell less, and have a higher tensile modulus, a higher yield stress and a lower dry-to-wet modulus ratio. It also predicts that disruption of the keratin matrix in water should lead to additional swelling, and a drop in modulus and yield stress. Our results are consistent with these predictions and suggest that the keratin matrix plays a critical role in governing the mechanical properties of mammalian keratins via control of IF hydration.

  16. Regulation of hard α-keratin mechanics via control of intermediate filament hydration: matrix squeeze revisited

    PubMed Central

    Greenberg, Daniel A.; Fudge, Douglas S.

    2013-01-01

    Mammalian hard α-keratins are fibre-reinforced biomaterials that consist of 10 nm intermediate filaments (IFs) embedded in an elastomeric protein matrix. Recent work suggests that the mechanical properties of IFs are highly sensitive to hydration, whereas hard α-keratins such as wool, hair and nail are relatively hydration insensitive. This raises the question of how mammalian keratins remain stiff in water. The matrix squeeze hypothesis states that the IFs in hard α-keratins are stiffened during an air-drying step during keratinization, and subsequently locked into a dehydrated state via the oxidation and cross-linking of the keratin matrix around them. The result is that even when hard α-keratins are immersed in water, their constituent IFs remain essentially ‘dry’ and therefore stiff. This hypothesis makes several predictions about the effects of matrix abundance and function on hard α-keratin mechanics and swelling behaviour. Specifically, it predicts that high matrix keratins in water will swell less, and have a higher tensile modulus, a higher yield stress and a lower dry-to-wet modulus ratio. It also predicts that disruption of the keratin matrix in water should lead to additional swelling, and a drop in modulus and yield stress. Our results are consistent with these predictions and suggest that the keratin matrix plays a critical role in governing the mechanical properties of mammalian keratins via control of IF hydration. PMID:23135675

  17. Controlling skeletal muscle CPT-I malonyl-CoA sensitivity: the importance of AMPK-independent regulation of intermediate filaments during exercise.

    PubMed

    Miotto, Paula M; Steinberg, Gregory R; Holloway, Graham P

    2017-02-15

    The obligatory role of carnitine palmitoyltransferase-I (CPT-I) in mediating mitochondrial lipid transport is well established, a process attenuated by malonyl-CoA (M-CoA). However, the necessity of reducing M-CoA concentrations to promote lipid oxidation has recently been challenged, suggesting external regulation on CPT-I. Since previous work in hepatocytes suggests the involvement of the intermediate filament fraction of the cytoskeleton in regulating CPT-I, we investigated in skeletal muscle if CPT-I sensitivity for M-CoA inhibition could be regulated by the intermediate filaments, and whether AMP-activated protein kinase (AMPK) could be involved in this process. Chemical disruption (3,3'-iminodipropionitrile, IDPN) of the intermediate filaments did not alter mitochondrial respiration or sensitivity for numerous substrates (palmitoyl-CoA, ADP, palmitoyl carnitine and pyruvate). In contrast, IDPN reduced CPT-I sensitivity for M-CoA inhibition in permeabilized muscle fibers, identifying M-CoA kinetics as a specific target for intermediate filament regulation. Importantly, exercise mimicked the effect of IDPN on M-CoA sensitivity, suggesting that intermediate filament disruption in vivo is physiologically important for CPT-I regulation. To ascertain a potential mechanism, since AMPK is activated during exercise, AMPK β1β2-KO mice were utilized in an attempt to ablate the observed exercise response. Unexpectedly, these mice displayed drastic attenuation in resting M-CoA sensitivity, such that exercise and IDPN could not further alter M-CoA sensitivity. These data suggest that AMPK is not required for the regulation of the intermediate filament interaction with CPT-I. Altogether, these data highlight that M-CoA sensitivity is important for regulating mitochondrial lipid transport. Moreover, M-CoA sensitivity appears to be regulated by intermediate filament interaction with CPT-I, a process that is important when metabolic homeostasis is challenged. © 2017 The

  18. Maintenance of electrostatic stabilization in altered tubulin lateral contacts may facilitate formation of helical filaments in foraminifera

    PubMed Central

    Bassen, David M.; Hou, Yubo; Bowser, Samuel S.; Banavali, Nilesh K.

    2016-01-01

    Microtubules in foraminiferan protists (forams) can convert into helical filament structures, in which longitudinal intraprotofilament interactions between tubulin heterodimers are thought to be lost, while lateral contacts across protofilaments are still maintained. The coarse geometric features of helical filaments are known through low-resolution negative stain electron microscopy (EM). In this study, geometric restraints derived from these experimental data were used to generate an average atomic-scale helical filament model, which anticipated a modest reorientation in the lateral tubulin heterodimer interface. Restrained molecular dynamics (MD) simulations of the nearest neighbor interactions combined with a Genalized Born implicit solvent model were used to assess the lateral, longitudinal, and seam contacts in 13-3 microtubules and the reoriented lateral contacts in the helical filament model. This electrostatic analysis suggests that the change in the lateral interface in the helical filament does not greatly diminish the lateral electrostatic interaction. After longitudinal dissociation, the 13-3 seam interaction is much weaker than the reoriented lateral interface in the helical filament model, providing a plausible atomic-detail explanation for seam-to-lateral contact transition that enables the transition to a helical filament structure. PMID:27539392

  19. Maintenance of electrostatic stabilization in altered tubulin lateral contacts may facilitate formation of helical filaments in foraminifera

    NASA Astrophysics Data System (ADS)

    Bassen, David M.; Hou, Yubo; Bowser, Samuel S.; Banavali, Nilesh K.

    2016-08-01

    Microtubules in foraminiferan protists (forams) can convert into helical filament structures, in which longitudinal intraprotofilament interactions between tubulin heterodimers are thought to be lost, while lateral contacts across protofilaments are still maintained. The coarse geometric features of helical filaments are known through low-resolution negative stain electron microscopy (EM). In this study, geometric restraints derived from these experimental data were used to generate an average atomic-scale helical filament model, which anticipated a modest reorientation in the lateral tubulin heterodimer interface. Restrained molecular dynamics (MD) simulations of the nearest neighbor interactions combined with a Genalized Born implicit solvent model were used to assess the lateral, longitudinal, and seam contacts in 13-3 microtubules and the reoriented lateral contacts in the helical filament model. This electrostatic analysis suggests that the change in the lateral interface in the helical filament does not greatly diminish the lateral electrostatic interaction. After longitudinal dissociation, the 13-3 seam interaction is much weaker than the reoriented lateral interface in the helical filament model, providing a plausible atomic-detail explanation for seam-to-lateral contact transition that enables the transition to a helical filament structure.

  20. Maintenance of electrostatic stabilization in altered tubulin lateral contacts may facilitate formation of helical filaments in foraminifera.

    PubMed

    Bassen, David M; Hou, Yubo; Bowser, Samuel S; Banavali, Nilesh K

    2016-08-19

    Microtubules in foraminiferan protists (forams) can convert into helical filament structures, in which longitudinal intraprotofilament interactions between tubulin heterodimers are thought to be lost, while lateral contacts across protofilaments are still maintained. The coarse geometric features of helical filaments are known through low-resolution negative stain electron microscopy (EM). In this study, geometric restraints derived from these experimental data were used to generate an average atomic-scale helical filament model, which anticipated a modest reorientation in the lateral tubulin heterodimer interface. Restrained molecular dynamics (MD) simulations of the nearest neighbor interactions combined with a Genalized Born implicit solvent model were used to assess the lateral, longitudinal, and seam contacts in 13-3 microtubules and the reoriented lateral contacts in the helical filament model. This electrostatic analysis suggests that the change in the lateral interface in the helical filament does not greatly diminish the lateral electrostatic interaction. After longitudinal dissociation, the 13-3 seam interaction is much weaker than the reoriented lateral interface in the helical filament model, providing a plausible atomic-detail explanation for seam-to-lateral contact transition that enables the transition to a helical filament structure.

  1. The 7-stranded structure of relaxed scallop muscle myosin filaments: support for a common head configuration in myosin-regulated muscles.

    PubMed

    Al-Khayat, Hind A; Morris, Edward P; Squire, John M

    2009-05-01

    Isolated relaxed myosin filaments from the myosin-regulated scallop striated adductor muscle have been reconstructed using electron microscopy and single particle analysis of negatively stained filaments. Three-dimensional reconstruction using 7-fold rotational symmetry but without imposed helical symmetry confirmed that the myosin head array is a 7-stranded, right-handed long-pitch 24/1 helix (or left-handed short-pitch 10/1 helix) with the whole structure having an axial repeat of 1440A. Reconstruction using the full helical symmetry revealed details of the myosin head density distribution within the head crowns in the relaxed scallop myosin filament. The resulting density distribution can best be explained by an arrangement in which the two heads from the same myosin molecule interact together within each crown in a compact parallel fashion along the filament axis. The configuration is consistent with the published configuration of the two heads within vertebrate smooth muscle myosin molecules observed in two-dimensional crystals of smooth muscle myosin and in the structure of tarantula myosin filaments. All these three muscle types are myosin-regulated, providing further support for a general motif of intramolecular interacting-heads structure in the relaxed state of myosin-regulated muscles as was proposed earlier by Woodhead et al. [Woodhead, J.L., Zhao, F.-Q., Craig, R., Egelman, E.H., Alamo, L., Padron, R.. 2005. Atomic model of a myosin filament in the relaxed state. Nature 436, 1195-1199]. However, the orientation of the Wendt structure is different from that found by Woodhead in that the outer head projects outwards and the inner head lies closer to the filament backbone, as in earlier work done on the insect flight muscle myosin filaments [AL-Khayat, H.A., Hudson, L., Reedy, M.K., Irving, T.C., Squire, J.M., 2003. Myosin head configuration in relaxed insect flight muscle: X-ray modelled resting crossbridges in a pre-powerstroke state are poised for

  2. Role of toxic and bioactive secondary metabolites in colonization and bloom formation by filamentous cyanobacteria Planktothrix.

    PubMed

    Kurmayer, Rainer; Deng, Li; Entfellner, Elisabeth

    2016-04-01

    Bloom-forming cyanobacteria Planktothrix agardhii and P. rubescens are regularly involved in the occurrence of cyanotoxin in lakes and reservoirs. Besides microcystins (MCs), which inhibit eukaryotic protein phosphatase 1 and 2A, several families of bioactive peptides are produced, thereby resulting in impressive secondary metabolite structural diversity. This review will focus on the current knowledge of the phylogeny, morphology, and ecophysiological adaptations of Planktothrix as well as the toxins and bioactive peptides produced. The relatively well studied ecophysiological adaptations (buoyancy, shade tolerance, nutrient storage capacity) can partly explain the invasiveness of this group of cyanobacteria that bloom within short periods (weeks to months). The more recent elucidation of the genetic basis of toxin and bioactive peptide synthesis paved the way for investigating its regulation both in the laboratory using cell cultures as well as under field conditions. The high frequency of several toxin and bioactive peptide synthesis genes observed within P. agardhii and P. rubescens, but not for other Planktothrix species (e.g. P. pseudagardhii), suggests a potential functional linkage between bioactive peptide production and the colonization potential and possible dominance in habitats. It is hypothesized that, through toxin and bioactive peptide production, Planktothrix act as a niche constructor at the ecosystem scale, possibly resulting in an even higher ability to monopolize resources, positive feedback loops, and resilience under stable environmental conditions. Thus, refocusing harmful algal bloom management by integrating ecological and phylogenetic factors acting on toxin and bioactive peptide synthesis gene distribution and concentrations could increase the predictability of the risks originating from Planktothrix blooms.

  3. A nerve growth factor-regulated messenger RNA encodes a new intermediate filament protein

    PubMed Central

    1988-01-01

    Differential screening of a cDNA library from the PC12 rat pheochromocytoma cell line previously revealed a clone, clone 73, whose corresponding mRNA is induced by nerve growth factor (NGF). Induction parallels NGF-stimulated PC12 differentiation from a chromaffinlike phenotype to a sympathetic neuronlike phenotype. We report that DNA sequence analysis reveals that clone 73 mRNA encodes an intermediate filament (IF) protein whose predicted amino acid sequence is distinct from the known sequences of other members of the IF protein family. The sequence has highest homology with desmin and vimentin and includes the highly conserved central alpha-helical rod domain with the characteristic heptad repeat of hydrophobic residues, but has lower homology in the amino-terminal head and carboxyl-terminal tail domains. The head domain contains a large number of serine residues which are potential phosphorylation sites. The expression of clone 73 in vivo in the nervous system of the adult rat was investigated by in situ hybridization of clone 73 probes to tissue sections. The mRNA is expressed at high levels in ganglia of the peripheral nervous system, including the superior cervical ganglion (sympathetic), ciliary ganglion (parasympathetic), and dorsal root ganglion (sensory). In the central nervous system, motor nuclei of cranial nerves III, IV, V, VI, VII, X, and XII as well as ventral horn motor neurons and a restricted set of other central nervous system nuclei express the clone 73 mRNA. Tissues apart from those of the nervous system did not in general express the mRNA, with only very low levels detected in adrenal gland. We discuss the implications of these results for the mechanism of NGF- induced PC12 cell differentiation, the pathways of neuronal development in vivo, and the possible function of the clone 73 IF protein and its relationship to other IF proteins. PMID:3339087

  4. Long 5' untranslated regions regulate the RNA stability of the deep-sea filamentous phage SW1.

    PubMed

    Jian, Huahua; Xiong, Lei; Xu, Guanpeng; Xiao, Xiang; Wang, Fengping

    2016-02-22

    Virus production in the deep-sea environment has been found to be high, and viruses have been suggested to play significant roles in the overall functioning of this ecosystem. Nevertheless, little is known about these viruses, including the mechanisms that control their production, which makes them one of the least understood biological entities on Earth. Previously, we isolated the filamentous phage SW1, whose virus production and gene transcription were found to be active at low temperatures, from a deep-sea bacterium, Shewanella piezotolerans WP3. In this study, the operon structure of phage SW1 is presented, which shows two operons with exceptionally long 5' and 3' untranslated regions (UTRs). In addition, the 5'UTR was confirmed to significantly influence the RNA stability of the SW1 transcripts. Our study revealed novel regulation of the operon and led us to propose a unique regulatory mechanism for Inoviruses. This type of RNA-based regulation may represent a mechanism for significant viral production in the cold deep biosphere.

  5. Long 5′ untranslated regions regulate the RNA stability of the deep-sea filamentous phage SW1

    PubMed Central

    Jian, Huahua; Xiong, Lei; Xu, Guanpeng; Xiao, Xiang; Wang, Fengping

    2016-01-01

    Virus production in the deep-sea environment has been found to be high, and viruses have been suggested to play significant roles in the overall functioning of this ecosystem. Nevertheless, little is known about these viruses, including the mechanisms that control their production, which makes them one of the least understood biological entities on Earth. Previously, we isolated the filamentous phage SW1, whose virus production and gene transcription were found to be active at low temperatures, from a deep-sea bacterium, Shewanella piezotolerans WP3. In this study, the operon structure of phage SW1 is presented, which shows two operons with exceptionally long 5′ and 3′ untranslated regions (UTRs). In addition, the 5′UTR was confirmed to significantly influence the RNA stability of the SW1 transcripts. Our study revealed novel regulation of the operon and led us to propose a unique regulatory mechanism for Inoviruses. This type of RNA-based regulation may represent a mechanism for significant viral production in the cold deep biosphere. PMID:26898180

  6. Formation of filament and plasma channel by the Bessel incident beam in Ar gas: role of the outer part of the beam.

    PubMed

    Song, Zhenming; Nakajima, Takashi

    2010-06-07

    We theoretically investigate the formation of filament and plasma channel in Ar gas by intense femtosecond pulses in the Bessel, truncated Bessel, and combination of two Gaussian modes. Through the numerical results obtained by solving the generalized nonlinear Schrödinger equation coupled with the electron density evolution equation, we find that there is a radial energy flow during the propagation, which implies that the outer part of the Bessel beam serves as an energy reservoir for the filament formed around the central peak. The results we obtain for the Bessel and truncated Bessel incident beams are consistent in that we can obtain a longer filament and plasma channel if more energy is reserved in the outer part of the Bessel incident beam. More interestingly we show that the combined use of two Gaussian beams with different beam diameters increases the energy stored in the outer part of the beam, and as a result the lengths of the filament and plasma channel become remarkably longer. This can be a practical choice to improve the propagation properties.

  7. Arabidopsis ACTIN-DEPOLYMERIZING FACTOR7 Severs Actin Filaments and Regulates Actin Cable Turnover to Promote Normal Pollen Tube Growth[W

    PubMed Central

    Zheng, Yiyan; Xie, Yurong; Jiang, Yuxiang; Qu, Xiaolu; Huang, Shanjin

    2013-01-01

    Actin filaments are often arranged into higher-order structures, such as the longitudinal actin cables that generate the reverse fountain cytoplasmic streaming pattern present in pollen tubes. While several actin binding proteins have been implicated in the generation of these cables, the mechanisms that regulate their dynamic turnover remain largely unknown. Here, we show that Arabidopsis thaliana ACTIN-DEPOLYMERIZING FACTOR7 (ADF7) is required for turnover of longitudinal actin cables. In vitro biochemical analyses revealed that ADF7 is a typical ADF that prefers ADP-G-actin over ATP-G-actin. ADF7 inhibits nucleotide exchange on actin and severs filaments, but its filament severing and depolymerizing activities are less potent than those of the vegetative ADF1. ADF7 primarily decorates longitudinal actin cables in the shanks of pollen tubes. Consistent with this localization pattern, the severing frequency and depolymerization rate of filaments significantly decreased, while their maximum lifetime significantly increased, in adf7 pollen tube shanks. Furthermore, an ADF7–enhanced green fluorescent protein fusion with defective severing activity but normal G-actin binding activity could not complement adf7, providing compelling evidence that the severing activity of ADF7 is vital for its in vivo functions. These observations suggest that ADF7 evolved to promote turnover of longitudinal actin cables by severing actin filaments in pollen tubes. PMID:24058157

  8. MamK, a bacterial actin, forms dynamic filaments in vivo that are regulated by the acidic proteins MamJ and LimJ

    PubMed Central

    Draper, Olga; Byrne, Meghan E.; Li, Zhuo; Keyhani, Sepehr; Cueto Barrozo, Joyce; Jensen, Grant; Komeili, Arash

    2011-01-01

    SUMMARY Bacterial actins, in contrast to their eukaryotic counterparts, are highly divergent proteins whose wide-ranging functions are thought to correlate with their evolutionary diversity. One clade, represented by the MamK protein of magnetotactic bacteria, is required for the subcellular organization of magnetosomes, membrane-bound organelles that aid in navigation along the earth’s magnetic field. Using a fluorescence recovery after photobleaching assay in Magnetospirillum magneticum AMB-1, we find that, like traditional actins, MamK forms dynamic filaments that require an intact NTPase motif for their turnover in vivo. We also uncover two proteins, MamJ and LimJ, which perform a redundant function to promote the dynamic behavior of MamK filaments in wildtype cells. The absence of both MamJ and LimJ leads to static filaments, a disrupted magnetosome chain, and an anomalous build-up of cytoskeletal filaments between magnetosomes. Our results suggest that MamK filaments, like eukaryotic actins, are intrinsically stable and rely on regulators for their dynamic behavior, a feature that stands in contrast to some classes of bacterial actins characterized to date. PMID:21883528

  9. 20 CFR 632.3 - Format for these regulations.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... NATIVE AMERICAN EMPLOYMENT AND TRAINING PROGRAMS Introduction § 632.3 Format for these regulations... with part 636 contains all the regulations under the Act applicable to Indian and Native American...

  10. 20 CFR 632.3 - Format for these regulations.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... NATIVE AMERICAN EMPLOYMENT AND TRAINING PROGRAMS Introduction § 632.3 Format for these regulations... with part 636 contains all the regulations under the Act applicable to Indian and Native American...

  11. 20 CFR 632.3 - Format for these regulations.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... NATIVE AMERICAN EMPLOYMENT AND TRAINING PROGRAMS Introduction § 632.3 Format for these regulations... with part 636 contains all the regulations under the Act applicable to Indian and Native American...

  12. The AMP-Related Kinase (AMPK) Induces Ca(2+)-Independent Dilation of Resistance Arteries by Interfering With Actin Filament Formation.

    PubMed

    Schubert, Kai Michael; Qiu, Jiehua; Blodow, Stephanie; Wiedenmann, Margarethe; Lubomirov, Lubomir T; Pfitzer, Gabriele; Pohl, Ulrich; Schneider, Holger

    2017-07-07

    Decreasing Ca(2+) sensitivity of vascular smooth muscle (VSM) allows for vasodilation without lowering of cytosolic Ca(2+). This may be particularly important in states requiring maintained dilation, such as hypoxia. AMP-related kinase (AMPK) is an important cellular energy sensor in VSM. Regulation of Ca(2+) sensitivity usually is attributed to myosin light chain phosphatase activity, but findings in non-VSM identified changes in the actin cytoskeleton. The potential role of AMPK in this setting is widely unknown. To assess the influence of AMPK on the actin cytoskeleton in VSM of resistance arteries with regard to potential Ca(2+) desensitization of VSM contractile apparatus. AMPK induced a slowly developing dilation at unchanged cytosolic Ca(2+) levels in potassium chloride-constricted intact arteries isolated from mouse mesenteric tissue. This dilation was not associated with changes in phosphorylation of myosin light chain or of myosin light chain phosphatase regulatory subunit. Using ultracentrifugation and confocal microscopy, we found that AMPK induced depolymerization of F-actin (filamentous actin). Imaging of arteries from LifeAct mice showed F-actin rarefaction in the midcellular portion of VSM. Immunoblotting revealed that this was associated with activation of the actin severing factor cofilin. Coimmunoprecipitation experiments indicated that AMPK leads to the liberation of cofilin from 14-3-3 protein. AMPK induces actin depolymerization, which reduces vascular tone and the response to vasoconstrictors. Our findings demonstrate a new role of AMPK in the control of actin cytoskeletal dynamics, potentially allowing for long-term dilation of microvessels without substantial changes in cytosolic Ca(2+). © 2017 American Heart Association, Inc.

  13. Arabidopsis RIC1 Severs Actin Filaments at the Apex to Regulate Pollen Tube Growth

    PubMed Central

    Zhou, Zhenzhen; Shi, Haifan; Chen, Binqing; Zhang, Ruihui; Huang, Shanjin; Fu, Ying

    2015-01-01

    Pollen tubes deliver sperms to the ovule for fertilization via tip growth. The rapid turnover of F-actin in pollen tube tips plays an important role in this process. In this study, we demonstrate that Arabidopsis thaliana RIC1, a member of the ROP-interactive CRIB motif-containing protein family, regulates pollen tube growth via its F-actin severing activity. Knockout of RIC1 enhanced pollen tube elongation, while overexpression of RIC1 dramatically reduced tube growth. Pharmacological analysis indicated that RIC1 affected F-actin dynamics in pollen tubes. In vitro biochemical assays revealed that RIC1 directly bound and severed F-actin in the presence of Ca2+ in addition to interfering with F-actin turnover by capping F-actin at the barbed ends. In vivo, RIC1 localized primarily to the apical plasma membrane (PM) of pollen tubes. The level of RIC1 at the apical PM oscillated during pollen tube growth. The frequency of F-actin severing at the apex was notably decreased in ric1-1 pollen tubes but was increased in pollen tubes overexpressing RIC1. We propose that RIC1 regulates F-actin dynamics at the apical PM as well as the cytosol by severing F-actin and capping the barbed ends in the cytoplasm, establishing a novel mechanism that underlies the regulation of pollen tube growth. PMID:25804540

  14. The respiratory syncytial virus fusion protein targets to the perimeter of inclusion bodies and facilitates filament formation by a cytoplasmic tail-dependent mechanism.

    PubMed

    Baviskar, Pradyumna S; Hotard, Anne L; Moore, Martin L; Oomens, Antonius G P

    2013-10-01

    The human respiratory syncytial virus (HRSV) fusion (F) protein cytoplasmic tail (CT) and matrix (M) protein are key mediators of viral assembly, but the underlying mechanisms are poorly understood. A complementation assay was developed to systematically examine the role of the F protein CT in infectious virus production. The ability of F mutants with alanine substitutions in the CT to complement an F-null virus in generating infectious progeny was quantitated by flow cytometry. Two CT regions with impact on infectious progeny production were identified: residues 557 to 566 (CT-R1) and 569 to 572 (CT-R2). Substitutions in CT-R1 decreased infectivity by 40 to 85% and increased the level of F-induced cell-cell fusion but had little impact on assembly of viral surface filaments, which are believed to be virions. Substitutions in CT-R2, as well as deletion of the entire CT, abrogated infectious progeny production and impaired viral filament formation. However, CT-R2 mutations did not block but rather delayed the formation of viral filaments, which continued to form at a low rate and contained the viral M protein and nucleoprotein (N). Microscopy analysis revealed that substitutions in CT-R2 but not CT-R1 led to accumulation of M and F proteins within and at the perimeter of viral inclusion bodies (IBs), respectively. The accumulation of M and F at IBs and coincident strong decrease in filament formation and infectivity upon CT-R2 mutations suggest that F interaction with IBs is an important step in the virion assembly process and that CT residues 569 to 572 act to facilitate release of M-ribonucleoprotein complexes from IBs.

  15. Regulation of myosin IIA and filamentous actin during insulin-stimulated glucose uptake in 3T3-L1 adipocytes

    SciTech Connect

    Stall, Richard; Ramos, Joseph; Kent Fulcher, F.; Patel, Yashomati M.

    2014-03-10

    Insulin stimulated glucose uptake requires the colocalization of myosin IIA (MyoIIA) and the insulin-responsive glucose transporter 4 (GLUT4) at the plasma membrane for proper GLUT4 fusion. MyoIIA facilitates filamentous actin (F-actin) reorganization in various cell types. In adipocytes F-actin reorganization is required for insulin-stimulated glucose uptake. What is not known is whether MyoIIA interacts with F-actin to regulate insulin-induced GLUT4 fusion at the plasma membrane. To elucidate the relationship between MyoIIA and F-actin, we examined the colocalization of MyoIIA and F-actin at the plasma membrane upon insulin stimulation as well as the regulation of this interaction. Our findings demonstrated that MyoIIA and F-actin colocalized at the site of GLUT4 fusion with the plasma membrane upon insulin stimulation. Furthermore, inhibition of MyoII with blebbistatin impaired F-actin localization at the plasma membrane. Next we examined the regulatory role of calcium in MyoIIA-F-actin colocalization. Reduced calcium or calmodulin levels decreased colocalization of MyoIIA and F-actin at the plasma membrane. While calcium alone can translocate MyoIIA it did not stimulate F-actin accumulation at the plasma membrane. Taken together, we established that while MyoIIA activity is required for F-actin localization at the plasma membrane, it alone is insufficient to localize F-actin to the plasma membrane. - Highlights: • Insulin induces colocalization of MyoIIA and F-actin at the cortex in adipocytes. • MyoIIA is necessary but not sufficient to localize F-actin at the cell cortex. • MyoIIA-F-actin colocalization is regulated by calcium and calmodulin.

  16. TetR Family Regulator brpT Modulates Biofilm Formation in Streptococcus sanguinis

    PubMed Central

    Ge, Xiuchun; Tang, Madison; Elrami, Fadi

    2017-01-01

    Biofilms are a key component in bacterial communities providing protection and contributing to infectious diseases. However, mechanisms involved in S. sanguinis biofilm formation have not been clearly elucidated. Here, we report the identification of a novel S. sanguinis TetR repressor, brpT (Biofilm Regulatory Protein TetR), involved in biofilm formation. Deletion of brpT resulted in a significant increase in biofilm formation. Interestingly, the mutant accumulated more water soluble and water insoluble glucans in its biofilm compared to the wild-type and the complemented mutant. The brpT mutation led to an altered biofilm morphology and structure exhibiting a rougher appearance, uneven distribution with more filaments bound to the chains. RNA-sequencing revealed that gtfP, the only glucosyltransferase present in S. sanguinis, was significantly up-regulated. In agreement with these findings, we independently observed that deletion of gtfP in S. sanguinis led to reduced biofilm and low levels of water soluble and insoluble glucans. These results suggest that brpT is involved in the regulation of the gtfP-mediated exopolysaccharide synthesis and controls S. sanguinis biofilm formation. The deletion of brpT may have a potential therapeutic application in regulating S. sanguinis colonization in the oral cavity and the prevention of dental caries. PMID:28046010

  17. Activation of thin-filament-regulated muscle by calcium ion: considerations based on nearest-neighbor lattice statistics.

    PubMed Central

    Shiner, J S; Solaro, R J

    1982-01-01

    We discuss the activation of thin-filament-regulated muscles by calcium ion in terms of a qualitative model based on nearest-neighbor lattice statistics. For the most part, the model takes into account only the essential features of the phenomenon--that there must be an interaction between calcium adsorption to troponin and crossbridge reaction with actin for calcium ion to activate contraction and that the relevant stationary states are nonequilibrium ones. Even so, the model predicts the following features which are seen experimentally but have generally not been considered in previous models: (i) the relative activations of stationary-state isometric force and ATPase are not equal; (ii) in general, neither activation of force nor that of ATPase is proportional to calcium adsorption to the activating sites; and (iii) the slopes of the relations between the activations and the logarithm of the calcium ion concentration generally depend on the necessary interaction between calcium ion adsorption and crossbridge reaction with actin. Thus, these relations show cooperative effects even if these is no interaction between calcium adsorption sites. PMID:6956882

  18. Intracellular fibril formation, calcification, and enrichment of chaperones, cytoskeletal, and intermediate filament proteins in the adult hippocampus CA1 following neonatal exposure to the nonprotein amino acid BMAA.

    PubMed

    Karlsson, Oskar; Berg, Anna-Lena; Hanrieder, Jörg; Arnerup, Gunnel; Lindström, Anna-Karin; Brittebo, Eva B

    2015-03-01

    The environmental neurotoxin β-N-methylamino-L-alanine (BMAA) has been implicated in the etiology of neurodegenerative disease, and recent studies indicate that BMAA can be misincorporated into proteins. BMAA is a developmental neurotoxicant that can induce long-term learning and memory deficits, as well as regionally restricted neuronal degeneration and mineralization in the hippocampal CA1. The aim of the study was to characterize long-term changes (2 weeks to 6 months) further in the brain of adult rats treated neonatally (postnatal days 9-10) with BMAA (460 mg/kg) using immunohistochemistry (IHC), transmission electron microscopy, and laser capture microdissection followed by LC-MS/MS for proteomic analysis. The histological examination demonstrated progressive neurodegenerative changes, astrogliosis, microglial activation, and calcification in the hippocampal CA1 3-6 months after exposure. The IHC showed an increased staining for α-synuclein and ubiquitin in the area. The ultrastructural examination revealed intracellular deposition of abundant bundles of closely packed parallel fibrils in neurons, axons, and astrocytes of the CA1. Proteomic analysis of the affected site demonstrated an enrichment of chaperones (e.g., clusterin, GRP-78), cytoskeletal and intermediate filament proteins, and proteins involved in the antioxidant defense system. Several of the most enriched proteins (plectin, glial fibrillar acidic protein, vimentin, Hsp 27, and ubiquitin) are known to form complex astrocytic inclusions, so-called Rosenthal fibers, in the neurodegenerative disorder Alexander disease. In addition, TDP-43 and the negative regulator of autophagy, GLIPR-2, were exclusively detected. The present study demonstrates that neonatal exposure to BMAA may offer a novel model for the study of hippocampal fibril formation in vivo.

  19. Light-Induced Movements of Chloroplasts and Nuclei Are Regulated in Both Cp-Actin-Filament-Dependent and -Independent Manners in Arabidopsis thaliana

    PubMed Central

    2016-01-01

    Light-induced chloroplast movement and attachment to the plasma membrane are dependent on actin filaments. In Arabidopsis thaliana, the short actin filaments on the chloroplast envelope, cp-actin filaments, are essential for chloroplast movement and positioning. Furthermore, cp-actin-filament-mediated chloroplast movement is necessary for the strong-light-induced nuclear avoidance response. The proteins CHLOROPLAST UNUSUAL POSITIONING 1 (CHUP1), KINESIN-LIKE PROTEIN FOR ACTIN-BASED CHLOROPLAST MOVEMENT 1 (KAC1) and KAC2 are required for the generation and/or maintenance of cp-actin filaments in Arabidopsis. In land plants, CHUP1 and KAC family proteins play pivotal roles in the proper movement of chloroplasts and their attachment to the plasma membrane. Here, we report similar but distinct phenotypes in chloroplast and nuclear photorelocation movements between chup1 and kac1kac2 mutants. Measurement of chloroplast photorelocation movement indicated that kac1kac2, but not chup1, exhibited a clear strong-light-induced increase in leaf transmittance changes. The chloroplast movement in kac1kac2 depended on phototropin 2, CHUP1 and two other regulators for cp-actin filaments, PLASTID MOVEMENT IMPAIRED 1 and THRUMIN 1. Furthermore, kac1kac2 retained a weak but significant nuclear avoidance response although chup1 displayed a severe defect in the nuclear avoidance response. The kac1kac2chup1 triple mutant was completely defective in both chloroplast and nuclear avoidance responses. These results indicate that CHUP1 and the KACs function somewhat independently, but interdependently mediate both chloroplast and nuclear photorelocation movements. PMID:27310016

  20. Light-Induced Movements of Chloroplasts and Nuclei Are Regulated in Both Cp-Actin-Filament-Dependent and -Independent Manners in Arabidopsis thaliana.

    PubMed

    Suetsugu, Noriyuki; Higa, Takeshi; Gotoh, Eiji; Wada, Masamitsu

    2016-01-01

    Light-induced chloroplast movement and attachment to the plasma membrane are dependent on actin filaments. In Arabidopsis thaliana, the short actin filaments on the chloroplast envelope, cp-actin filaments, are essential for chloroplast movement and positioning. Furthermore, cp-actin-filament-mediated chloroplast movement is necessary for the strong-light-induced nuclear avoidance response. The proteins CHLOROPLAST UNUSUAL POSITIONING 1 (CHUP1), KINESIN-LIKE PROTEIN FOR ACTIN-BASED CHLOROPLAST MOVEMENT 1 (KAC1) and KAC2 are required for the generation and/or maintenance of cp-actin filaments in Arabidopsis. In land plants, CHUP1 and KAC family proteins play pivotal roles in the proper movement of chloroplasts and their attachment to the plasma membrane. Here, we report similar but distinct phenotypes in chloroplast and nuclear photorelocation movements between chup1 and kac1kac2 mutants. Measurement of chloroplast photorelocation movement indicated that kac1kac2, but not chup1, exhibited a clear strong-light-induced increase in leaf transmittance changes. The chloroplast movement in kac1kac2 depended on phototropin 2, CHUP1 and two other regulators for cp-actin filaments, PLASTID MOVEMENT IMPAIRED 1 and THRUMIN 1. Furthermore, kac1kac2 retained a weak but significant nuclear avoidance response although chup1 displayed a severe defect in the nuclear avoidance response. The kac1kac2chup1 triple mutant was completely defective in both chloroplast and nuclear avoidance responses. These results indicate that CHUP1 and the KACs function somewhat independently, but interdependently mediate both chloroplast and nuclear photorelocation movements.

  1. αE-catenin actin-binding domain alters actin filament conformation and regulates binding of nucleation and disassembly factors

    PubMed Central

    Hansen, Scott D.; Kwiatkowski, Adam V.; Ouyang, Chung-Yueh; Liu, HongJun; Pokutta, Sabine; Watkins, Simon C.; Volkmann, Niels; Hanein, Dorit; Weis, William I.; Mullins, R. Dyche; Nelson, W. James

    2013-01-01

    The actin-binding protein αE-catenin may contribute to transitions between cell migration and cell–cell adhesion that depend on remodeling the actin cytoskeleton, but the underlying mechanisms are unknown. We show that the αE-catenin actin-binding domain (ABD) binds cooperatively to individual actin filaments and that binding is accompanied by a conformational change in the actin protomer that affects filament structure. αE-catenin ABD binding limits barbed-end growth, especially in actin filament bundles. αE-catenin ABD inhibits actin filament branching by the Arp2/3 complex and severing by cofilin, both of which contact regions of the actin protomer that are structurally altered by αE-catenin ABD binding. In epithelial cells, there is little correlation between the distribution of αE-catenin and the Arp2/3 complex at developing cell–cell contacts. Our results indicate that αE-catenin binding to filamentous actin favors assembly of unbranched filament bundles that are protected from severing over more dynamic, branched filament arrays. PMID:24068324

  2. Contributions of EspA Filaments and Curli Fimbriae in Cellular Adherence and Biofilm Formation of Enterohemorrhagic Escherichia coli O157:H7

    PubMed Central

    Sharma, Vijay K.; Kudva, Indira T.; Bearson, Bradley L.; Stasko, Judith A.

    2016-01-01

    In Escherichia coli O157:H7 (O157), the filamentous structure of the type III secretion system is produced from the polymerization of the EspA protein. EspA filaments are essential for O157 adherence to epithelial cells. In previous studies, we demonstrated that O157 hha deletion mutants showed increased adherence to HEp-2 cells and produced abundant biofilms. Transcriptional analysis revealed increased expression of espA as well as the csgA gene, which encodes curli fimbriae that are essential for biofilm formation. In the present study, we constructed hha espA, hha csgA, and hha csgA espA deletion mutants to determine the relative importance of EspA and CsgA in O157 adherence to HEp-2 cells and biofilm formation. In vitro adherence assays, conducted at 37°C in a tissue culture medium containing 0.1% glucose, showed that HEp-2 cell adherence required EspA because hha espA and hha csgA espA mutants adhered to HEp-2 cells at higher levels only when complemented with an espA-expressing plasmid. Biofilm assays performed at 28°C in a medium lacking glucose showed dependency of biofilm formation on CsgA; however EspA was not produced under these conditions. Despite production of detectable levels of EspA at 37°C in media supplemented with 0.1% glucose, the biofilm formation occurred independent of EspA. These results indicate dependency of O157 adherence to epithelial cells on EspA filaments, while CsgA promoted biofilm formation under conditions mimicking those found in the environment (low temperature with nutrient limitations) and in the digestive tract of an host animal (higher temperature and low levels of glucose). PMID:26900701

  3. Cooperative regulation of myosin-S1 binding to actin filaments by a continuous flexible Tm-Tn chain.

    PubMed

    Mijailovich, Srboljub M; Kayser-Herold, Oliver; Li, Xiaochuan; Griffiths, Hugh; Geeves, Michael A

    2012-12-01

    The regulation of striated muscle contraction involves cooperative interactions between actin filaments, myosin-S1 (S1), tropomyosin (Tm), troponin (Tn), and calcium. These interactions are modeled by treating overlapping tropomyosins as a continuous flexible chain (CFC), weakly confined by electrostatic interactions with actin. The CFC is displaced locally in opposite directions on the actin surface by the binding of either S1 or Troponin I (TnI) to actin. The apparent rate constants for myosin and TnI binding to and detachment from actin are then intrinsically coupled via the CFC model to the presence of neighboring bound S1s and TnIs. Monte Carlo simulations at prescribed values of the CFC stiffness, the CFC's degree of azimuthal confinement, and the angular displacements caused by the bound proteins were able to predict the stopped-flow transients of S1 binding to regulated F-actin. The transients collected over a large range of calcium concentrations could be well described by adjusting a single calcium-dependent parameter, the rate constant of TnI detachment from actin, k(-I). The resulting equilibrium constant K(B) ≡ 1/K(I) varied sigmoidally with the free calcium, increasing from 0.12 at low calcium (pCa >7) to 12 at high calcium (pCa <5.5) with a Hill coefficient of ~2.15. The similarity of the curves for excess-actin and excess-myosin data confirms their allosteric relationship. The spatially explicit calculations confirmed variable sizes for the cooperative units and clustering of bound myosins at low calcium concentrations. Moreover, inclusion of negative cooperativity between myosin units predicted the observed slowing of myosin binding at excess-myosin concentrations.

  4. Suppression subtractive hybridization and comparative expression analysis to identify developmentally regulated genes in filamentous fungi.

    PubMed

    Gesing, Stefan; Schindler, Daniel; Nowrousian, Minou

    2013-09-01

    Ascomycetes differentiate four major morphological types of fruiting bodies (apothecia, perithecia, pseudothecia and cleistothecia) that are derived from an ancestral fruiting body. Thus, fruiting body differentiation is most likely controlled by a set of common core genes. One way to identify such genes is to search for genes with evolutionary conserved expression patterns. Using suppression subtractive hybridization (SSH), we selected differentially expressed transcripts in Pyronema confluens (Pezizales) by comparing two cDNA libraries specific for sexual and for vegetative development, respectively. The expression patterns of selected genes from both libraries were verified by quantitative real time PCR. Expression of several corresponding homologous genes was found to be conserved in two members of the Sordariales (Sordaria macrospora and Neurospora crassa), a derived group of ascomycetes that is only distantly related to the Pezizales. Knockout studies with N. crassa orthologues of differentially regulated genes revealed a functional role during fruiting body development for the gene NCU05079, encoding a putative MFS peptide transporter. These data indicate conserved gene expression patterns and a functional role of the corresponding genes during fruiting body development; such genes are candidates of choice for further functional analysis.

  5. Laser-filament-induced snow formation in a subsaturated zone in a cloud chamber: experimental and theoretical study.

    PubMed

    Ju, Jingjing; Sun, Haiyi; Sridharan, Aravindan; Wang, Tie-Jun; Wang, Cheng; Liu, Jiansheng; Li, Ruxin; Xu, Zhizhan; Chin, See Leang

    2013-12-01

    1 kHz, 2 mJ, 45 fs, 800 nm laser pulses were fired into a laboratory diffusion cloud chamber through a subsaturated zone (relative humidity ∼73%, T ∼ 4.3 °C). After 60 min of laser irradiation, an oval-shaped snow pile was observed right below the filament center and weighed ∼12.0 mg. The air current velocity at the edge of the vortices was estimated to be ∼16.5 cm/s. Scattering scenes recorded from the side show that filament-induced turbulence were formed inside the cloud chamber with two vortices below the filament. Two-dimensional simulations of the air flow motion in two cross sections of the cloud chamber confirm that the turbulent vortices exist below the filament. Based upon this simulation, we deduce that the vortices indeed have a three-dimensional elliptical shape. Hence, we propose that inside vortices where the humidity was supersaturated or saturated the condensation nuclei, namely, HNO(3), N(2)(+), O(2)(+) and other aerosols and impurities, were activated and grew in size. Large-sized particles would eventually be spun out along the fast moving direction towards the cold plate and formed an oval-shaped snow pile at the end.

  6. Filamentation and spatiotemporal distribution of extracellular polymeric substances: role on X.fastidiosa single cell adhesion and biofilm formation (Conference Presentation)

    NASA Astrophysics Data System (ADS)

    Janissen, Richard; Murillo, Duber M.; Niza, Barbara; Sahoo, Prasana K.; Monteiro, Moniellen P.; César, Carlos L.; Carvalho, Hernandes F.; de Souza, Alessandra A.; Cotta, Monica A.

    2016-04-01

    Biofilms can be defined as a community of microorganisms attached to a surface, living embedded in a self- produced matrix of hydrated extracellular polymeric substances (EPS) which comprises most of the biofilm mass. We have recently used an extensive pool of microscopy techniques (confocal fluorescence, electron and scanning probe microscopies) at the micro and nanoscales in order to create a detailed temporal observation of Xylella fastidiosa biofilm formation, using both wild type strain and Green Fluorescent Protein (GFP)-modified cells of this citrus phytopathogen. We have identified three different EPS compositions, as well as their spatial and temporal distribution from single cell to mature biofilm formation stages. In the initial adhesion stage, soluble-EPS (S-EPS) accumulates at cell polar regions and forms a surface layer which facilitates irreversible cell attachment and cell cluster formation. These small clusters are subsequently connected by filamentous cells; further S-EPS surface coverage facilitates cell attachment and form filaments, leading to a floating framework of mature biofilms. The important role of EPS in X.fastidiosa biology was further investigated by imunolabelling experiments to detect the distribution of XadA1 adhesin, which is expressed in early stages of biofilm formation and released in outer membrane vesicles. This protein is located mainly in S-EPS covered areas, as well as on the filaments, indicating a molecular pathway to the enhanced cell attachment previously observed. These results suggest that S-EPS may thus represent an important target for disease control, slow plant colonization by the bacteria, keeping the plant more productive in the field.

  7. Outflow Feedback Regulated Massive Star Formation in Parsec-Scale Cluster Forming Clumps

    SciTech Connect

    Wang, Peng; Li, Zhi-Yun; Abel, Tom; Nakamura, Fumitaka; /Niigata U.

    2010-02-15

    We investigate massive star formation in turbulent, magnetized, parsec-scale clumps of molecular clouds including protostellar outflow feedback using three dimensional numerical simulations of effective resolution 2048{sup 3}. The calculations are carried out using a block structured adaptive mesh refinement code that solves the ideal MHD equations including self-gravity and implements accreting sink particles. We find that, in the absence of regulation by magnetic fields and outflow feedback, massive stars form readily in a turbulent, moderately condensed clump of {approx} 1,600 M{sub {circle_dot}} (containing {approx} 10{sup 2} initial Jeans masses), along with a cluster of hundreds of lower mass stars. The massive stars are fed at high rates by (1) transient dense filaments produced by large-scale turbulent compression at early times, and (2) by the clump-wide global collapse resulting from turbulence decay at late times. In both cases, the bulk of the massive star's mass is supplied from outside a 0.1 pc-sized 'core' that surrounds the star. In our simulation, the massive star is clump-fed rather than core-fed. The need for large-scale feeding makes the massive star formation prone to regulation by outflow feedback, which directly opposes the feeding processes. The outflows reduce the mass accretion rates onto the massive stars by breaking up the dense filaments that feed the massive star formation at early times, and by collectively slowing down the global collapse that fuel the massive star formation at late times. The latter is aided by a moderate magnetic field of strength in the observed range (corresponding to a dimensionless clump mass-to-flux ratio {lambda} {approx} a few); the field allows the outflow momenta to be deposited more efficiently inside the clump. We conclude that the massive star formation in our simulated turbulent, magnetized, parsec-scale clump is outflow-regulated and clump-fed (ORCF for short). An important implication is that the

  8. THE EFFECT OF ENVIRONMENT ON THE FORMATION OF H{alpha} FILAMENTS AND COOL CORES IN GALAXY GROUPS AND CLUSTERS

    SciTech Connect

    McDonald, Michael; Veilleux, Sylvain; Mushotzky, Richard E-mail: veilleux@astro.umd.edu

    2011-04-10

    We present the results of a combined X-ray and H{alpha} study of 10 galaxy groups and 17 galaxy clusters using the Chandra X-ray Observatory and the Maryland Magellan Tunable Filter. We find no difference in the morphology or detection frequency of H{alpha} filaments in groups versus clusters over the mass range 10{sup 13} < M{sub 500} < 10{sup 15} M{sub sun}. The detection frequency of H{alpha} emission is shown to be only weakly dependent on the total mass of the system at the 52% confidence level. In contrast, we find that the presence of H{alpha} filaments is strongly correlated with both the global (89% confidence level) and core (84%) intracluster medium (ICM) entropy, as well as the X-ray cooling rate (72%). The H{alpha} filaments are therefore an excellent proxy for the cooling ICM. The H{alpha} filaments are more strongly correlated with the cooling properties of the ICM than with the radio properties of the brightest cluster galaxy; this further supports the scenario where these filaments are directly associated with a thermally unstable, rapidly cooling ICM, rather than radio bubbles. The ICM cooling efficiency, defined as the X-ray cooling rate per unit gas mass, is shown to correlate with the total system mass, indicating that groups are more efficient at cooling than clusters. This result implies that, in systems with cool cores, active galactic nucleus feedback scales with the total mass of the system, in agreement with earlier suggestions.

  9. Signaling molecules regulating phenotypic conversions of astrocytes and glial scar formation in damaged nerve tissues.

    PubMed

    Koyama, Yutaka

    2014-12-01

    Phenotypic conversion of astrocytes from resting to reactive (i.e., astrocytic activation) occurs in numerous brain disorders. Astrocytic activation in severely damaged brain regions often leads to glial scar formation. Because astrocytic activation and glial scar largely affect the vulnerability and tissue repair of damaged brain, numerous studies have been made to clarify mechanisms regulating the astrocytic phenotype. The phenotypic conversion is accompanied by the increased expression of intermediate filament proteins and the induction of hypertrophy in reactive astrocytes. Severe brain damage results in proliferation and migration of reactive astrocytes, which lead to glial scar formations at the injured areas. Gliogenesis from neural progenitors in the adult brain is also involved in astrocytic activation and glial scar formation. Recent studies have shown that increased expression of connexin 43, aquaporin 4, matrix metalloproteinase 9, and integrins alter the function of astrocytes. The transcription factors: STAT3, OLIG2, SMAD, NF-κB, and Sp1 have been suggested to play regulatory roles in astrocytic activation and glial scar formation. In this review, I discuss the roles of these key molecules regulating the pathophysiological functions of reactive astrocytes. Copyright © 2014 Elsevier Ltd. All rights reserved.

  10. Metabolomics protocols for filamentous fungi.

    PubMed

    Gummer, Joel P A; Krill, Christian; Du Fall, Lauren; Waters, Ormonde D C; Trengove, Robert D; Oliver, Richard P; Solomon, Peter S

    2012-01-01

    Proteomics and transcriptomics are established functional genomics tools commonly used to study filamentous fungi. Metabolomics has recently emerged as another option to complement existing techniques and provide detailed information on metabolic regulation and secondary metabolism. Here, we describe broad generic protocols that can be used to undertake metabolomics studies in filamentous fungi.

  11. Epigenetic Regulation of Memory Formation and Maintenance

    ERIC Educational Resources Information Center

    Zovkic, Iva B.; Guzman-Karlsson, Mikael C.; Sweatt, J. David

    2013-01-01

    Understanding the cellular and molecular mechanisms underlying the formation and maintenance of memories is a central goal of the neuroscience community. It is well regarded that an organism's ability to lastingly adapt its behavior in response to a transient environmental stimulus relies on the central nervous system's capability for structural…

  12. Formative Justice: The Regulative Principle of Education

    ERIC Educational Resources Information Center

    McClintock, Robert

    2016-01-01

    Background/Context: Concepts of justice relevant to making personal and public decisions about education. Purpose: To clarify a concept of formative justice that persons and the public often ignore in making decisions about educational effort. Setting: "The windmills of your mind" Research Design: Reflective essay.…

  13. Epigenetic Regulation of Memory Formation and Maintenance

    ERIC Educational Resources Information Center

    Zovkic, Iva B.; Guzman-Karlsson, Mikael C.; Sweatt, J. David

    2013-01-01

    Understanding the cellular and molecular mechanisms underlying the formation and maintenance of memories is a central goal of the neuroscience community. It is well regarded that an organism's ability to lastingly adapt its behavior in response to a transient environmental stimulus relies on the central nervous system's capability for structural…

  14. Formative Justice: The Regulative Principle of Education

    ERIC Educational Resources Information Center

    McClintock, Robert

    2016-01-01

    Background/Context: Concepts of justice relevant to making personal and public decisions about education. Purpose: To clarify a concept of formative justice that persons and the public often ignore in making decisions about educational effort. Setting: "The windmills of your mind" Research Design: Reflective essay.…

  15. Role of Intermediate Filaments in Vesicular Traffic

    PubMed Central

    Margiotta, Azzurra; Bucci, Cecilia

    2016-01-01

    Intermediate filaments are an important component of the cellular cytoskeleton. The first established role attributed to intermediate filaments was the mechanical support to cells. However, it is now clear that intermediate filaments have many different roles affecting a variety of other biological functions, such as the organization of microtubules and microfilaments, the regulation of nuclear structure and activity, the control of cell cycle and the regulation of signal transduction pathways. Furthermore, a number of intermediate filament proteins have been involved in the acquisition of tumorigenic properties. Over the last years, a strong involvement of intermediate filament proteins in the regulation of several aspects of intracellular trafficking has strongly emerged. Here, we review the functions of intermediate filaments proteins focusing mainly on the recent knowledge gained from the discovery that intermediate filaments associate with key proteins of the vesicular membrane transport machinery. In particular, we analyze the current understanding of the contribution of intermediate filaments to the endocytic pathway. PMID:27120621

  16. The MAP kinase-activated protein kinase Rck2p regulates cellular responses to cell wall stresses, filamentation and virulence in the human fungal pathogen Candida albicans.

    PubMed

    Li, Xichuan; Du, Wei; Zhao, Jingwen; Zhang, Lilin; Zhu, Zhiyan; Jiang, Linghuo

    2010-06-01

    Rck2p is the Hog1p-MAP kinase-activated protein kinase required for the attenuation of protein synthesis in response to an osmotic challenge in Saccharomyces cerevisiae. Rck2p also regulates rapamycin sensitivity in both S. cerevisiae and Candida albicans. In this study, we demonstrate that the deletion of CaRCK2 renders C. albicans cells sensitive to, and CaRck2p translocates from the cytosol to the nucleus in response to, cell wall stresses caused by Congo red, Calcoflor White, elevated heat and zymolyase. However, the kinase activity of CaRck2p is not required for the cellular response to these cell wall stresses. Furthermore, transcripts of cell wall protein-encoding genes CaBGL2, CaHWP1 and CaXOG1 are reduced in C. albicans cells lacking CaRCK2. The deletion of CaRCK2 also reduces the in vitro filamentation of C. albicans and its virulence in a mouse model of systemic candidasis. The kinase activity of CaRck2p is required for the virulence, but not for the in vitro filamentation, in C. albicans. Therefore, Rck2p regulates cellular responses to cell wall stresses, filamentation and virulence in the human fungal pathogen C. albicans.

  17. CAPRICE positively regulates stomatal formation in the Arabidopsis hypocotyl.

    PubMed

    Serna, Laura

    2008-12-01

    In the Arabidopsis hypocotyl, stomata develop only from a set of epidermal cell files. Previous studies have identified several negative regulators of stomata formation. Such regulators also trigger non-hair cell fate in the root. Here, it is shown that TOO MANY MOUTHS (TMM) positively regulates CAPRICE (CPC) expression in differentiating stomaless-forming cell files, and that the CPC protein might move to the nucleus of neighbouring stoma-forming cells, where it promotes stomata formation in a redundant manner with TRIPTYCHON (TRY). Unexpectedly, the CPC protein was also localized in the nucleus and peripheral cytoplasm of hypocotyl fully differentiated epidermal cells, suggesting that CPC plays an additional role to those related to stomata formation. These results identify CPC and TRY as positive regulators of stomata formation in the embryonic stem, which increases the similarity between the genetic control of root hair and stoma cell fate determination.

  18. Magnetic Fields in Massive Filaments

    NASA Astrophysics Data System (ADS)

    Pillai, Thushara

    Magnetic fields pervade galaxies, shaping them from the largest scales to the smallest star forming scales. A firm understanding of their role is crucial to our understanding of the physics of ISM. A dominant phase of the ISM that has received considerable attention is that of filaments which are ubiquitous and dominate the mass reservoir in molecular clouds. Enormous progress has been made recently towards understanding filament properties. The next major step should be to understand the role of magnetic fields in filaments. We propose to take advantage of HAWC+ dust emission polarimeter now available on SOFIA to launch a pilot polarization study towards three major classes of filaments: (i) Pristine (ii) Hub-Filament systems and (iii) Perturbed. HAWC+ will trace the connection between the star forming cores and the filaments enveloping them. By covering a vast range in parameter space from quiescent to active filaments, we will be constraining the initial conditions prior to star formation, during star formation and after star formation (feedback from newly formed stars on their parent clouds.) The interpretation of observations will be supported by extensive custom-made numerical simulations of magnetized clouds and subsequent dust radiative transfer with various grain alignment mechanisms, as provided by collaborators. Combined, these observations will provide the first panoramic view of the magnetized nature of massive filaments in the ISM.

  19. Magnetic Fields in Massive Filaments

    NASA Astrophysics Data System (ADS)

    Pillai, G. S. Thushara

    2015-10-01

    Magnetic fields pervade galaxies, shaping them from the largest scales to the smallest star forming scales. A firm understanding of their role is crucial to our understanding of the physics of ISM. A dominant phase of the ISM that has received considerable attention is that of filaments which are ubiquitous and dominate the mass reservoir in molecular clouds. Enormous progress has been made recently towards understanding filament properties. The next major step should be to understand the role of magnetic fields in filaments. We propose to take advantage of HAWC+ dust emission polarimeter now available on SOFIA to launch a pilot polarization study towards three major classes of filaments: (i) Pristine (ii) Hub-Filament systems and (iii) Perturbed. HAWC+ will trace the connection between the star forming cores and the filaments enveloping them. By covering a vast range in parameter space from quiescent to active filaments, we will be constraining the initial conditions prior to star formation, during star formation and after star formation (feedback from newly formed stars on their parent clouds.) The interpretation of observations will be supported by extensive custom--made numerical simulations of magnetized clouds and subsequent dust radiative transfer with various grain alignment mechanisms, as provided by collaborators. Combined, these observations will provide the first panoramic view of the magnetized nature of massive filaments in the ISM.

  20. Filaments in Lupus I

    NASA Astrophysics Data System (ADS)

    Takahashi, Satoko; Rodon, J.; De Gregorio-Monsalvo, I.; Plunkett, A.

    2017-06-01

    The mechanisms behind the formation of sub-stellar mass sources are key to determine the populations at the low-mass end of the stellar distribution. Here, we present mapping observations toward the Lupus I cloud in C18O(2-1) and 13CO(2-1) obtained with APEX. We have identified a few velocity-coherent filaments. Each contains several substellar mass sources that are also identified in the 1.1mm continuum data (see also SOLA catalogue presentation). We will discuss the velocity structure, fragmentation properties of the identified filaments, and the nature of the detected sources.

  1. Formative Assessment: Assessment Is for Self-Regulated Learning

    ERIC Educational Resources Information Center

    Clark, Ian

    2012-01-01

    The article draws from 199 sources on assessment, learning, and motivation to present a detailed decomposition of the values, theories, and goals of formative assessment. This article will discuss the extent to which formative feedback actualizes and reinforces self-regulated learning (SRL) strategies among students. Theoreticians agree that SRL…

  2. Formative Assessment: Assessment Is for Self-Regulated Learning

    ERIC Educational Resources Information Center

    Clark, Ian

    2012-01-01

    The article draws from 199 sources on assessment, learning, and motivation to present a detailed decomposition of the values, theories, and goals of formative assessment. This article will discuss the extent to which formative feedback actualizes and reinforces self-regulated learning (SRL) strategies among students. Theoreticians agree that SRL…

  3. 20 CFR 633.103 - Format for these regulations.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 20 Employees' Benefits 3 2010-04-01 2010-04-01 false Format for these regulations. 633.103 Section 633.103 Employees' Benefits EMPLOYMENT AND TRAINING ADMINISTRATION, DEPARTMENT OF LABOR MIGRANT AND... applicable to migrant and other seasonally employed farmworker programs. (b) Should the regulations at...

  4. Cigarette Smoke-Related Hydroquinone Induces Filamentous Actin Reorganization and Heat Shock Protein 27 Phosphorylation through p38 and Extracellular Signal-Regulated Kinase 1/2 in Retinal Pigment Epithelium

    PubMed Central

    Pons, Marianne; Cousins, Scott W.; Csaky, Karl G.; Striker, Gary; Marin-Castaño, Maria E.

    2010-01-01

    Retinal pigment epithelium (RPE)-derived membranous debris named blebs, may accumulate and contribute to sub-RPE deposit formation, which is the earliest sign of age-related macular degeneration (AMD). Oxidative injury to the RPE might play a significant role in AMD. However, the underlying mechanisms are unknown. We previously reported that hydroquinone (HQ), a major pro-oxidant in cigarette smoke, foodstuff, and atmospheric pollutants, induces actin rearrangement and membrane blebbing in RPE cells as well as sub-RPE deposits in mice. Here, we show for the first time that phosphorylated Heat shock protein 27 (Hsp27), a key regulator of actin filaments dynamics, is up-regulated in RPE from patients with AMD. Also, HQ-induced nonlethal oxidative injury led to Hsp27mRNA up-regulation, dimer formation, and Hsp27 phosphorylation in ARPE-19 cells. Furthermore, we found that a cross talk between p38 and extracellular signal-regulated kinase (ERK) mediates HQ-induced Hsp27 phosphorylation and actin aggregate formation, revealing ERK as a novel upstream mediator of Hsp27 phosphorylation. Finally, we demonstrated that Hsp25, p38, and ERK phosphorylation are increased in aging C57BL/6 mice chronically exposed to HQ, whereas Hsp25 expression is decreased. Our data suggest that phosphorylated Hsp27 might be a key mediator in AMD and HQ-induced oxidative injury to the RPE, which may provide helpful insights into the early cellular events associated with actin reorganization and bleb formation involved in sub-RPE deposits formation relevant to the pathogenesis of AMD. PMID:20651235

  5. Exploring the Role of Filamentous Cyanobacteria in the Formation of Living Silica Stromatolites in Yellowstone Hot Springs

    NASA Astrophysics Data System (ADS)

    Stueeken, E. E.; Loyd, S.; Li, X.; Callow, R.; de La Torre, J.; Berelson, W.; Corsetti, F.; Hanselmann, K.; Stevenson, B. S.; Spear, J.; Pepe-Ranney, C.; Beer, L.; Dawson, S.; Johnson, H. A.; 2008, G.

    2008-12-01

    Ancient stromatolites of various morphologies occur in the geologic record. Their interpretation as being a cyanobacterial biosignature is controversial due to the phylogenetic diversity of microbes associated with modern stromatolites. In this study, we show that microstructures in siliceous living stromatolites are controlled by the preferential silica mineralization of cyanobacterial filaments. SEM images of siliceous stromatolites collected from a hot spring in Yellowstone National Park reveal that the 5-10cm-sized structures are composed of abundant microscopic tubes. The dark-light alternating laminations of the stromatolite are due to the orientation of tubes with respect to the surface of the stromatolite. Light layers are 150-200um thick and consist of tubes perpendicular to the surface whereas dark layers are 50-75um-thick and consist of tubes lying parallel. The light layers have greater porosity than the denser, dark layers; some of this porosity consists of large (200um diameter) pores around which the tubes bend. Tube diameters have a bimodal distribution with most being either 1µm or 2.8µm thick. The tube diameters are similar to the average cross- section of living filamentous cyanobacteria of the same hot spring. This suggests that the tubes are actually silicified filaments and that the stromatolite is a record of biological activity. Additionally, 16S libraries generated from the microbial communities living on and in the stromatolite show a diverse array of microorganisms. The community diversity is in stark contrast to the stromatolite's homogeneous micro- structural composition; the stromatolite appears to consist entirely of silicified tubes. Other microbial morphologies are not observed in SEM images. It appears that the cyanobacterial sheaths are preferentially silicified compared with other microbial surfaces. Although the microbial role in carbonate accretions may be quite different than in siliceous settings, these results corroborate

  6. Filamentous Influenza Viruses

    PubMed Central

    Badham, Matthew D.; Rossman, Jeremy S.

    2016-01-01

    Influenza A virus is a pathogen of global medical importance causing significant health and socio-economic costs every year. Influenza virus is an unusual pathogen in that it is pleomorphic, capable of forming virions ranging in shape from spherical to filamentous. Despite decades of research on the influenza virus, much remains unknown about the formation of filamentous influenza viruses and their role in the viral replication cycle. Here, we discuss what is known about influenza virus assembly and budding, focusing on the viral and host factors that are involved in the determination of viral morphology. Whilst the biological function of the filamentous morphology remains unknown, recent results suggest a role in facilitating viral spread in vivo. We discuss these results and speculate on the consequences of viral morphology during influenza virus infection of the human respiratory tract. PMID:28042529

  7. Filament Breakaway

    NASA Image and Video Library

    2017-04-18

    A dark, elongated filament rose up and broke to the lower left and out from the sun seen by NASA Solar Dynamics Observatory, Apr.9-10, 2017. Filaments are cooler clouds of plasma tethered above the sun surface by magnetic forces. They are notoriously unstable and tend not to last more than a few days before they collapse into the sun or break away into space. A video, taken in extreme ultraviolet light, covers about nine hours of activity. Videos are available at https://photojournal.jpl.nasa.gov/catalog/PIA21592

  8. Interplay between I308 and Y310 residues in the third repeat of microtubule-binding domain is essential for tau filament formation.

    PubMed

    Naruto, Keiko; Minoura, Katsuhiko; Okuda, Ryouhei; Taniguchi, Taizo; In, Yasuko; Ishida, Toshimasa; Tomoo, Koji

    2010-10-08

    Investigation of the mechanism of tau polymerization is indispensable for finding inhibitory conditions or identifying compounds preventing the formation of paired helical filament or oligomers. Tau contains a microtubule-binding domain consisting of three or four repeats in its C-terminal half. It has been considered that the key event in tau polymerization is the formation of a β-sheet structure arising from a short hexapeptide (306)VQIVYK(311) in the third repeat of tau. In this paper, we report for the first time that the C-H⋯π interaction between Ile308 and Tyr310 is the elemental structural scaffold essential for forming a dry "steric zipper" structure in tau amyloid fibrils.

  9. Regulation of biofilm formation in Pseudomonas and Burkholderia species.

    PubMed

    Fazli, Mustafa; Almblad, Henrik; Rybtke, Morten Levin; Givskov, Michael; Eberl, Leo; Tolker-Nielsen, Tim

    2014-07-01

    In the present review, we describe and compare the molecular mechanisms that are involved in the regulation of biofilm formation by Pseudomonas putida, Pseudomonas fluorescens, Pseudomonas aeruginosa and Burkholderia cenocepacia. Our current knowledge suggests that biofilm formation is regulated by cyclic diguanosine-5'-monophosphate (c-di-GMP), small RNAs (sRNA) and quorum sensing (QS) in all these bacterial species. The systems that employ c-di-GMP as a second messenger regulate the production of exopolysaccharides and surface proteins which function as extracellular matrix components in the biofilms formed by the bacteria. The systems that make use of sRNAs appear to regulate the production of exopolysaccharide biofilm matrix material in all these species. In the pseudomonads, QS regulates the production of extracellular DNA, lectins and biosurfactants which all play a role in biofilm formation. In B.cenocepacia QS regulates the expression of a large surface protein, lectins and extracellular DNA that all function as biofilm matrix components. Although the three regulatory systems all regulate the production of factors used for biofilm formation, the molecular mechanisms involved in transducing the signals into expression of the biofilm matrix components differ between the species. Under the conditions tested, exopolysaccharides appears to be the most important biofilm matrix components for P.aeruginosa, whereas large surface proteins appear to be the most important biofilm matrix components for P.putida, P.fluorescens, and B.cenocepacia.

  10. Tropomyosin-dependent filament formation by a polymerization-defective mutant yeast actin (V266G,L267G).

    PubMed

    Wen, K K; Kuang, B; Rubenstein, P A

    2000-12-22

    A major function of tropomyosin (TPM) in nonmuscle cells may be stabilization of F-actin by binding longitudinally along the actin filament axis. However, no clear evidence exists in vitro that TPM can significantly affect the critical concentration of actin. We previously made a polymerization-defective mutant actin, GG (V266G, L267G). This actin will not polymerize alone at 25 degrees C but will in the presence of phalloidin or beryllium fluoride. With beryllium fluoride, but not phalloidin, this polymerization rescue is cold-sensitive. We show here that GG-actin polymerizability was restored by cardiac tropomyosin and yeast TPM1 and TPM2 at 25 degrees C with rescue efficiency inversely proportional to TPM length (TPM2 > TPM1 > cardiac tropomyosin), indicating the importance of the ends in polymerization rescue. In the presence of TPM, the apparent critical concentration of actin is 5.5 microm, 10-15-fold higher than that of wild type actin but well below that of the GG-actin alone (>20 microm). Non N-acetylated TPMs did not rescue GG-actin polymerization. The TPMs did not prevent cold-induced depolymerization of GG F-actin. TPM-dependent GG-actin polymerization did not occur at temperatures below 20 degrees C. Polymerization rescue may depend initially on the capture of unstable GG-F-actin oligomers by the TPM, resulting in the strengthening of actin monomer-monomer contacts along the filament axis.

  11. Interactions with Actin Monomers, Actin Filaments, and Arp2/3 Complex Define the Roles of WASP Family Proteins and Cortactin in Coordinately Regulating Branched Actin Networks*

    PubMed Central

    Helgeson, Luke A.; Prendergast, Julianna G.; Wagner, Andrew R.; Rodnick-Smith, Max; Nolen, Brad J.

    2014-01-01

    Arp2/3 complex is an important actin filament nucleator that creates branched actin filament networks required for formation of lamellipodia and endocytic actin structures. Cellular assembly of branched actin networks frequently requires multiple Arp2/3 complex activators, called nucleation promoting factors (NPFs). We recently presented a mechanism by which cortactin, a weak NPF, can displace a more potent NPF, N-WASP, from nascent branch junctions to synergistically accelerate nucleation. The distinct roles of these NPFs in branching nucleation are surprising given their similarities. We biochemically dissected these two classes of NPFs to determine how their Arp2/3 complex and actin interacting segments modulate their influences on branched actin networks. We find that the Arp2/3 complex-interacting N-terminal acidic sequence (NtA) of cortactin has structural features distinct from WASP acidic regions (A) that are required for synergy between the two NPFs. Our mutational analysis shows that differences between NtA and A do not explain the weak intrinsic NPF activity of cortactin, but instead that cortactin is a weak NPF because it cannot recruit actin monomers to Arp2/3 complex. We use TIRF microscopy to show that cortactin bundles branched actin filaments using actin filament binding repeats within a single cortactin molecule, but that N-WASP antagonizes cortactin-mediated bundling. Finally, we demonstrate that multiple WASP family proteins synergistically activate Arp2/3 complex and determine the biochemical requirements in WASP proteins for synergy. Our data indicate that synergy between WASP proteins and cortactin may play a general role in assembling diverse actin-based structures, including lamellipodia, podosomes, and endocytic actin networks. PMID:25160634

  12. Mechanosensitive store-operated calcium entry regulates the formation of cell polarity.

    PubMed

    Huang, Yi-Wei; Chang, Shu-Jing; I-Chen Harn, Hans; Huang, Hui-Ting; Lin, Hsi-Hui; Shen, Meng-Ru; Tang, Ming-Jer; Chiu, Wen-Tai

    2015-09-01

    Ca(2+) -mediated formation of cell polarity is essential for directional migration which plays an important role in physiological and pathological processes in organisms. To examine the critical role of store-operated Ca(2+) entry, which is the major form of extracellular Ca(2+) influx in non-excitable cells, in the formation of cell polarity, we employed human bone osteosarcoma U2OS cells, which exhibit distinct morphological polarity during directional migration. Our analyses showed that Ca(2+) was concentrated at the rear end of cells and that extracellular Ca(2+) influx was important for cell polarization. Inhibition of store-operated Ca(2+) entry using specific inhibitors disrupted the formation of cell polarity in a dose-dependent manner. Moreover, the channelosomal components caveolin-1, TRPC1, and Orai1 were concentrated at the rear end of polarized cells. Knockdown of TRPC1 or a TRPC inhibitor, but not knockdown of Orai1, reduced cell polarization. Furthermore, disruption of lipid rafts or overexpression of caveolin-1 contributed to the downregulation of cell polarity. On the other hand, we also found that cell polarity, store-operated Ca(2+) entry activity, and cell stiffness were markedly decreased by low substrate rigidity, which may be caused by the disorganization of actin filaments and microtubules that occurs while regulating the activity of the mechanosensitive TRPC1 channel.

  13. Dynamic localization of HmpF regulates type IV pilus activity and directional motility in the filamentous cyanobacterium Nostoc punctiforme.

    PubMed

    Cho, Ye Won; Gonzales, Alfonso; Harwood, Thomas V; Huynh, Jessica; Hwang, Yeji; Park, Jun Sang; Trieu, Anthony Q; Italia, Parth; Pallipuram, Vivek K; Risser, Douglas D

    2017-10-01

    Many cyanobacteria exhibit surface motility powered by type 4 pili (T4P). In the model filamentous cyanobacterium Nostoc punctiforme, the T4P systems are arrayed in static, bipolar rings in each cell. The chemotaxis-like Hmp system is essential for motility and the coordinated polar accumulation of PilA on cells in motile filaments, while the Ptx system controls positive phototaxis. Using transposon mutagenesis, a gene, designated hmpF, was identified as involved in motility. Synteny among filamentous cyanobacteria and the similar expression patterns for hmpF and hmpD imply that HmpF is part of the Hmp system. Deletion of hmpF produced a phenotype distinct from other hmp genes, but indistinguishable from pilB or pilQ. Both an HmpF-GFPuv fusion protein, and PilA, as assessed by in situ immunofluorescence, displayed coordinated, unipolar localization at the leading pole of each cell. Reversals were modulated by changes in light intensity and preceded by the migration of HmpF-GFPuv to the lagging cell poles. These results are consistent with a model where direct interaction between HmpF and the T4P system activates pilus extension, the Hmp system facilitates coordinated polarity of HmpF to establish motility, and the Ptx system modulates HmpF localization to initiate reversals in response to changes in light intensity. © 2017 John Wiley & Sons Ltd.

  14. Actin filament organization in activated mast cells is regulated by heterotrimeric and small GTP-binding proteins

    PubMed Central

    1994-01-01

    Rat peritoneal mast cells, both intact and permeabilized, have been used widely as model secretory cells. GTP-binding proteins and calcium play a major role in controlling their secretory response. Here we have examined changes in the organization of actin filaments in intact mast cells after activation by compound 48/80, and in permeabilized cells after direct activation of GTP-binding proteins by GTP-gamma-S. In both cases, a centripetal redistribution of cellular F-actin was observed: the content of F-actin was reduced in the cortical region and increased in the cell interior. The overall F-actin content was increased. Using permeabilized cells, we show that AIF4-, an activator of heterotrimeric G proteins, induces the disassembly of F-actin at the cortex, while the appearance of actin filaments in the interior of the cell is dependent on two small GTPases, rho and rac. Rho was found to be responsible for de novo actin polymerization, presumably from a membrane-bound monomeric pool, while rac was required for an entrapment of the released cortical filaments. Thus, a heterotrimeric G-protein and the small GTPases, rho and rac, participate in affecting the changes in the actin cytoskeleton observed after activation of mast cells. PMID:8051203

  15. Regulation of protein phosphorylation of the intermediate-sized filament vimentin in the ciliary epithelium of the mammalian eye

    SciTech Connect

    Coca-Prados, M.

    1985-08-25

    The intermediate-sized filaments of vimentin-type (Mr = 57,000) have been identified biochemically and immunochemically as a major cytoskeleton component in the ciliary epithelium of the mammalian eye. When human or rabbit ciliary processes, or cultured ciliary epithelial-derived cells were incubated in serum-free medium containing (TSP)orthophosphate and any of the following agents: 1) beta-adrenergic agonists (isoproterenol or epinephrine), 2) direct activators of adenylate cyclase (cholera toxin or forskolin), 3) analogs of cyclic AMP (8-Br-cAMP), or 4) prostaglandin E1, the phosphorylation of vimentin was significantly enhanced. The maximal enhancement ranged, in vivo and in vitro, from about 3-fold in human to 5-fold in rabbit, with either 1 mM 8-Br-cAMP or 0.1 microM forskolin. Indirect immunofluorescence microscopy using a monoclonal antibody, anti-vimentin, allowed the localization of vimentin filaments in cultured ciliary epithelial cells. Treatment of these cells in culture with the catecholamine hormone, isoproterenol (1 microM), resulted in a profound reorganization of vimentin filaments. This may be correlated with the enhanced levels of phosphorylated vimentin observed upon increasing cellular cyclic AMP.

  16. Filament winding

    NASA Astrophysics Data System (ADS)

    Shibley, A. M.

    The major aspects of filament winding are discussed, emphasizing basic reinforcement and matrix materials, winding procedures, process controls, and cured composite properties. Fiberglass (E-glass and S-glass strengths are 500,000 and 665,000 psi respectively) and polyester resins are the principal reinforcement constituent materials. Graphite and aramid reinforcements are being used more frequently, primarily for the more critical pressure vessels. Matrix systems are most commonly based on epoxy as it has superior mechanical properties, fatigue behavior, and heat resistance as compard with polyesters. A fiberglass overwrap of PVC pipe is an anticipated development in on-site winding and combination winding, and the compression molding of filament wound lay-ups will be investigated. The fabrication of weight-sensitive structural components may be achieved by using such moldings.

  17. H-treatment impact on conductive-filament formation and stability in Ta{sub 2}O{sub 5}-based resistive-switching memory cells

    SciTech Connect

    Goux, L. Redolfi, A.; Jurczak, M.; Kim, J. Y.; Magyari-Kope, B.; Nishi, Y.

    2015-03-28

    In this article, we evidence the lower formation energy and improved stability of the conductive filament (CF) formed in TiN\\Ta{sub 2}O{sub 5}\\Ta resistive-switching memory cells treated in NH{sub 3} atmosphere at 400 °C. This annealing treatment results in (i) lower forming voltage, (ii) lower CF resistance, and (iii) longer retention lifetime of the oxygen-vacancy (V{sub o}) chain constituting the CF. Atomistic insights into these processes are provided by ab initio calculations performed for hydrogen (H) species incorporated in non-stoichiometric Ta{sub 2}O{sub 5} supercells: (i) V{sub o} formation energy is reduced by the presence of H, (ii) V{sub o}-chain CF conductivity is increased by V{sub o} + OH complex formation, and (iii) V{sub o}-chain retention is strengthened by the stable V{sub o} + OH complex. As a result, efficient CF formation and excellent state stability are obtained after 15 days at 250 °C.

  18. Similar is not the same: differences in the function of the (hemi-)cellulolytic regulator XlnR (Xlr1/Xyr1) in filamentous fungi.

    PubMed

    Klaubauf, Sylvia; Narang, Hari Mander; Post, Harm; Zhou, Miaomiao; Brunner, Kurt; Mach-Aigner, Astrid R; Mach, Robert L; Heck, Albert J R; Altelaar, A F Maarten; de Vries, Ronald P

    2014-11-01

    The transcriptional activator XlnR (Xlr1/Xyr1) is a major regulator in fungal xylan and cellulose degradation as well as in the utilization of d-xylose via the pentose catabolic pathway. XlnR homologs are commonly found in filamentous ascomycetes and often assumed to have the same function in different fungi. However, a comparison of the saprobe Aspergillus niger and the plant pathogen Magnaporthe oryzae showed different phenotypes for deletion strains of XlnR. In this study wild type and xlnR/xlr1/xyr1 mutants of five fungi were compared: Fusarium graminearum, M. oryzae, Trichoderma reesei, A. niger and Aspergillus nidulans. Growth profiling on relevant substrates and a detailed analysis of the secretome as well as extracellular enzyme activities demonstrated a common role of this regulator in activating genes encoding the main xylanolytic enzymes. However, large differences were found in the set of genes that is controlled by XlnR in the different species, resulting in the production of different extracellular enzyme spectra by these fungi. This comparison emphasizes the functional diversity of a fine-tuned (hemi-)cellulolytic regulatory system in filamentous fungi, which might be related to the adaptation of fungi to their specific biotopes. Data are available via ProteomeXchange with identifier PXD001190. Copyright © 2014 Elsevier Inc. All rights reserved.

  19. The control of cellular shape and motility. Mg2+ and tropomyosin regulate the formation and the dissociation of microfilament bundles.

    PubMed Central

    Grazi, E; Cuneo, P; Cataldi, A

    1992-01-01

    At pH 7.14 and 37 degrees C, in 7.2% (w/v) poly(ethylene glycol) 6000, tropomyosin-regulated actin filaments are converted into filament bundles by increasing the free Mg2+ concentration to 1.7-2.0 mM. When free Mg2+ concentration is decreased below 1.7 mM, bundles dissociate back into tropomyosin-regulated actin filaments. Pure actin filaments are insensitive to this mechanism of control and are found as filament bundles in all the range of free Mg2+ concentrations tested (1.37-2.2 mM). The mechanism of regulation described above is likely to operate in the cell, where the concentration of free Mg2+ is linked to the energy charge of the adenine nucleotide system. Images Fig. 3. PMID:1471985

  20. Transcriptional regulation of secondary growth and wood formation.

    PubMed

    Du, Juan; Groover, Andrew

    2010-01-01

    Secondary growth and wood formation are products of the vascular cambium, a lateral meristem. Although the mechanisms have only recently begun to be uncovered, transcriptional regulation appears increasingly central to the regulation of secondary growth. The importance of transcriptional regulation is illustrated by the correlation of expression of specific classes of genes with related biological processes occurring at specific stages of secondary growth, including cell division, cell expansion, and cell differentiation. At the same time, transcription factors have been characterized that affect specific aspects of secondary growth, including regulation of the cambium and differentiation of cambial daughter cells. In the present review, we summarize evidence pointing to transcription as a major mechanism for regulation of secondary growth, and outline future approaches for comprehensively describing transcriptional networks underlying secondary growth.

  1. Effects of amino acid on morphological development and nucleus formation of arachidonic acid-producing filamentous micro-organism, Mortierella alpina.

    PubMed

    Koizumi, K; Higashiyama, K; Park, E Y

    2006-04-01

    Effects of amino acid on morphological development and nucleus formation of arachidonic acid-producing filamentous micro-organism, Mortierella alpina were investigated using flow-through chamber. Mortierella alpina CBS 754.68 was cultivated in flow through chamber using nutrient-rich, minimal and specific amino acid-containing minimal media. To investigate the effect of amino acid on morphological parameters either 0.28 g l(-1) alanine, 0.53 g l(-1) sodium glutamate one hydrate or 0.42 g l(-1) valine was added to the minimal medium. In a flow-through chamber, the growth of hyphal elements and nucleus formation of arachidonic acid-producing fungus M. alpina were studied on-line, using image analysis techniques. When the Ala- and Val-containing media were used, the hyphal growth units (HGUs) were 90.2 and 86.7 microm per tip, respectively, which were 2.4-fold higher than that in the nutrient-rich medium, indicating that Ala and Val stimulate the elongation of hyphae. The specific nucleus formation rates were Glu->Val-containing media>minimal and nutrient-rich media>Ala-containing medium. The nucleus doubling times in Glu- and Val-containing media were 1.9 and 2 h, respectively, which were not significant different. Ala and Val stimulate the elongation of M. alpina hyphae, and nucleus formation rates were Glu->Val->Ala-containing media. Formation of fungal morphology and nucleus were shown using the flow-through chamber coupled with image analysis, which making possible to discuss the relationship between mycelial morphology and nucleus formation of M. alpina.

  2. Leptin regulates bone formation via the sympathetic nervous system

    NASA Technical Reports Server (NTRS)

    Takeda, Shu; Elefteriou, Florent; Levasseur, Regis; Liu, Xiuyun; Zhao, Liping; Parker, Keith L.; Armstrong, Dawna; Ducy, Patricia; Karsenty, Gerard

    2002-01-01

    We previously showed that leptin inhibits bone formation by an undefined mechanism. Here, we show that hypothalamic leptin-dependent antiosteogenic and anorexigenic networks differ, and that the peripheral mediators of leptin antiosteogenic function appear to be neuronal. Neuropeptides mediating leptin anorexigenic function do not affect bone formation. Leptin deficiency results in low sympathetic tone, and genetic or pharmacological ablation of adrenergic signaling leads to a leptin-resistant high bone mass. beta-adrenergic receptors on osteoblasts regulate their proliferation, and a beta-adrenergic agonist decreases bone mass in leptin-deficient and wild-type mice while a beta-adrenergic antagonist increases bone mass in wild-type and ovariectomized mice. None of these manipulations affects body weight. This study demonstrates a leptin-dependent neuronal regulation of bone formation with potential therapeutic implications for osteoporosis.

  3. Leptin regulates bone formation via the sympathetic nervous system

    NASA Technical Reports Server (NTRS)

    Takeda, Shu; Elefteriou, Florent; Levasseur, Regis; Liu, Xiuyun; Zhao, Liping; Parker, Keith L.; Armstrong, Dawna; Ducy, Patricia; Karsenty, Gerard

    2002-01-01

    We previously showed that leptin inhibits bone formation by an undefined mechanism. Here, we show that hypothalamic leptin-dependent antiosteogenic and anorexigenic networks differ, and that the peripheral mediators of leptin antiosteogenic function appear to be neuronal. Neuropeptides mediating leptin anorexigenic function do not affect bone formation. Leptin deficiency results in low sympathetic tone, and genetic or pharmacological ablation of adrenergic signaling leads to a leptin-resistant high bone mass. beta-adrenergic receptors on osteoblasts regulate their proliferation, and a beta-adrenergic agonist decreases bone mass in leptin-deficient and wild-type mice while a beta-adrenergic antagonist increases bone mass in wild-type and ovariectomized mice. None of these manipulations affects body weight. This study demonstrates a leptin-dependent neuronal regulation of bone formation with potential therapeutic implications for osteoporosis.

  4. A Novel Small Molecule Inhibitor of Candida albicans Biofilm Formation, Filamentation and Virulence with Low Potential for the Development of Resistance

    PubMed Central

    Pierce, Christopher G.; Chaturvedi, Ashok K.; Lazzell, Anna L.; Powell, Alexander T.; Saville, Stephen. P.; McHardy, Stanton F.; Lopez-Ribot, Jose L.

    2015-01-01

    Background/Objectives Candida albicans is the principal causative agent of candidiasis, the most common fungal infection in humans. Candidiasis represents the third-to-fourth most frequent nosocomial infection worldwide, as this normal commensal of humans causes opportunistic infections in an expanding population of immune- and medically-compromised patients. These infections are frequently associated with biofilm formation, which complicates treatment and contributes to unacceptably high mortality rates. Methods To address the pressing need for new antifungals we have performed a high content screen of 20,000 small molecules in a chemical library (NOVACore™) to identify compounds that inhibit C. albicans biofilm formation, and conducted a series of follow-up studies to examine the in vitro and in vivo activity of the identified compounds. Results The screen identified a novel series of diazaspiro-decane structural analogs which were largely represented among the bioactive compounds. Characterization of the leading compound from this series indicated that it inhibits processes associated with C. albicans virulence, most notably biofilm formation and filamentation, without having an effect on overall growth or eliciting resistance. This compound demonstrated in vivo activity in clinically-relevant murine models of both invasive and oral candidiasis and as such represents a promising lead for antifungal drug development. Furthermore, these results provide proof of concept for the implementation of anti-virulence approaches against C. albicans and other fungal infections that would be less likely to foster the emergence of resistance. PMID:26691764

  5. Cyclic diguanylate regulation of Bacillus cereus group biofilm formation.

    PubMed

    Fagerlund, Annette; Smith, Veronika; Røhr, Åsmund K; Lindbäck, Toril; Parmer, Marthe P; Andersson, K Kristoffer; Reubsaet, Leon; Økstad, Ole Andreas

    2016-08-01

    Biofilm formation can be considered a bacterial virulence mechanism. In a range of Gram-negatives, increased levels of the second messenger cyclic diguanylate (c-di-GMP) promotes biofilm formation and reduces motility. Other bacterial processes known to be regulated by c-di-GMP include cell division, differentiation and virulence. Among Gram-positive bacteria, where the function of c-di-GMP signalling is less well characterized, c-di-GMP was reported to regulate swarming motility in Bacillus subtilis while having very limited or no effect on biofilm formation. In contrast, we show that in the Bacillus cereus group c-di-GMP signalling is linked to biofilm formation, and to several other phenotypes important to the lifestyle of these bacteria. The Bacillus thuringiensis 407 genome encodes eleven predicted proteins containing domains (GGDEF/EAL) related to c-di-GMP synthesis or breakdown, ten of which are conserved through the majority of clades of the B. cereus group, including Bacillus anthracis. Several of the genes were shown to affect biofilm formation, motility, enterotoxin synthesis and/or sporulation. Among these, cdgF appeared to encode a master diguanylate cyclase essential for biofilm formation in an oxygenated environment. Only two cdg genes (cdgA, cdgJ) had orthologs in B. subtilis, highlighting differences in c-di-GMP signalling between B. subtilis and B. cereus group bacteria.

  6. DELLA proteins regulate arbuscule formation in arbuscular mycorrhizal symbiosis.

    PubMed

    Floss, Daniela S; Levy, Julien G; Lévesque-Tremblay, Véronique; Pumplin, Nathan; Harrison, Maria J

    2013-12-17

    Most flowering plants are able to form endosymbioses with arbuscular mycorrhizal fungi. In this mutualistic association, the fungus colonizes the root cortex and establishes elaborately branched hyphae, called arbuscules, within the cortical cells. Arbuscule development requires the cellular reorganization of both symbionts, and the resulting symbiotic interface functions in nutrient exchange. A plant symbiosis signaling pathway controls the development of the symbiosis. Several components of the pathway have been identified, but transcriptional regulators that control downstream pathways for arbuscule formation are still unknown. Here we show that DELLA proteins, which are repressors of gibberellic acid (GA) signaling and function at the nexus of several signaling pathways, are required for arbuscule formation. Arbuscule formation is severely impaired in a Medicago truncatula Mtdella1/Mtdella2 double mutant; GA treatment of wild-type roots phenocopies the della double mutant, and a dominant DELLA protein (della1-Δ18) enables arbuscule formation in the presence of GA. Ectopic expression of della1-Δ18 suggests that DELLA activity in the vascular tissue and endodermis is sufficient to enable arbuscule formation in the inner cortical cells. In addition, expression of della1-Δ18 restores arbuscule formation in the symbiosis signaling pathway mutant cyclops/ipd3, indicating an intersection between DELLA and symbiosis signaling for arbuscule formation. GA signaling also influences arbuscule formation in monocots, and a Green Revolution wheat variety carrying dominant DELLA alleles shows enhanced colonization but a limited growth response to arbuscular mycorrhizal symbiosis.

  7. DELLA proteins regulate arbuscule formation in arbuscular mycorrhizal symbiosis

    PubMed Central

    Floss, Daniela S.; Levy, Julien G.; Lévesque-Tremblay, Véronique; Pumplin, Nathan; Harrison, Maria J.

    2013-01-01

    Most flowering plants are able to form endosymbioses with arbuscular mycorrhizal fungi. In this mutualistic association, the fungus colonizes the root cortex and establishes elaborately branched hyphae, called arbuscules, within the cortical cells. Arbuscule development requires the cellular reorganization of both symbionts, and the resulting symbiotic interface functions in nutrient exchange. A plant symbiosis signaling pathway controls the development of the symbiosis. Several components of the pathway have been identified, but transcriptional regulators that control downstream pathways for arbuscule formation are still unknown. Here we show that DELLA proteins, which are repressors of gibberellic acid (GA) signaling and function at the nexus of several signaling pathways, are required for arbuscule formation. Arbuscule formation is severely impaired in a Medicago truncatula Mtdella1/Mtdella2 double mutant; GA treatment of wild-type roots phenocopies the della double mutant, and a dominant DELLA protein (della1-Δ18) enables arbuscule formation in the presence of GA. Ectopic expression of della1-Δ18 suggests that DELLA activity in the vascular tissue and endodermis is sufficient to enable arbuscule formation in the inner cortical cells. In addition, expression of della1-Δ18 restores arbuscule formation in the symbiosis signaling pathway mutant cyclops/ipd3, indicating an intersection between DELLA and symbiosis signaling for arbuscule formation. GA signaling also influences arbuscule formation in monocots, and a Green Revolution wheat variety carrying dominant DELLA alleles shows enhanced colonization but a limited growth response to arbuscular mycorrhizal symbiosis. PMID:24297892

  8. A keratin scaffold regulates epidermal barrier formation, mitochondrial lipid composition, and activity

    PubMed Central

    Kumar, Vinod; Bouameur, Jamal-Eddine; Bär, Janina; Rice, Robert H.; Hornig-Do, Hue-Tran; Roop, Dennis R.; Schwarz, Nicole; Brodesser, Susanne; Thiering, Sören; Leube, Rudolf E.; Wiesner, Rudolf J.; Vijayaraj, Preethi; Brazel, Christina B.; Heller, Sandra; Binder, Hans; Löffler-Wirth, Henry; Seibel, Peter

    2015-01-01

    Keratin intermediate filaments (KIFs) protect the epidermis against mechanical force, support strong adhesion, help barrier formation, and regulate growth. The mechanisms by which type I and II keratins contribute to these functions remain incompletely understood. Here, we report that mice lacking all type I or type II keratins display severe barrier defects and fragile skin, leading to perinatal mortality with full penetrance. Comparative proteomics of cornified envelopes (CEs) from prenatal KtyI−/− and KtyII−/−K8 mice demonstrates that absence of KIF causes dysregulation of many CE constituents, including downregulation of desmoglein 1. Despite persistence of loricrin expression and upregulation of many Nrf2 targets, including CE components Sprr2d and Sprr2h, extensive barrier defects persist, identifying keratins as essential CE scaffolds. Furthermore, we show that KIFs control mitochondrial lipid composition and activity in a cell-intrinsic manner. Therefore, our study explains the complexity of keratinopathies accompanied by barrier disorders by linking keratin scaffolds to mitochondria, adhesion, and CE formation. PMID:26644517

  9. Moesin regulates stable microtubule formation and limits retroviral infection in cultured cells.

    PubMed

    Naghavi, Mojgan H; Valente, Susana; Hatziioannou, Theodora; de Los Santos, Kenia; Wen, Ying; Mott, Christina; Gundersen, Gregg G; Goff, Stephen P

    2007-01-10

    In a functional screen of mammalian complementary DNA libraries, we identified moesin as a novel gene whose overexpression blocks infection by murine leukemia viruses and human immunodeficiency virus type 1 in human and rodent lines, before the initiation of reverse transcription. Knockdown of moesin by RNA interference resulted in enhanced infection, suggesting that even the endogenous basal levels of moesin in rat fibroblasts are sufficient to limit virus infection. Moesin acts as a crosslinker between plasma membrane and actin filaments, as well as a signal transducer in responses involving cytoskeletal remodeling. Moesin overexpression was found to downregulate the formation of stable microtubules, whereas knockdown of moesin increased stable microtubule formation. A virus-resistant mutant cell line also displayed decreased stable microtubule levels, and virus-sensitive revertants recovered from the mutant line showed restoration of the stable microtubules, suggesting that these cytoskeletal networks play an important role in early post-entry events in the retroviral lifecycle. Together, these results suggest that moesin negatively regulates stable microtubule networks and is a natural determinant of cellular sensitivity to retroviral infection.

  10. nArgBP2 regulates excitatory synapse formation by controlling dendritic spine morphology.

    PubMed

    Lee, Sang-Eun; Kim, Yoonju; Han, Jeong-Kyu; Park, Hoyong; Lee, Unghwi; Na, Myeongsu; Jeong, Soomin; Chung, ChiHye; Cestra, Gianluca; Chang, Sunghoe

    2016-06-14

    Neural Abelson-related gene-binding protein 2 (nArgBP2) was originally identified as a protein that directly interacts with synapse-associated protein 90/postsynaptic density protein 95-associated protein 3 (SAPAP3), a postsynaptic scaffolding protein critical for the assembly of glutamatergic synapses. Although genetic deletion of nArgBP2 in mice leads to manic/bipolar-like behaviors resembling many aspects of symptoms in patients with bipolar disorder, the actual function of nArgBP2 at the synapse is completely unknown. Here, we found that the knockdown (KD) of nArgBP2 by specific small hairpin RNAs (shRNAs) resulted in a dramatic change in dendritic spine morphology. Reintroducing shRNA-resistant nArgBP2 reversed these defects. In particular, nArgBP2 KD impaired spine-synapse formation such that excitatory synapses terminated mostly at dendritic shafts instead of spine heads in spiny neurons, although inhibitory synapse formation was not affected. nArgBP2 KD further caused a marked increase of actin cytoskeleton dynamics in spines, which was associated with increased Wiskott-Aldrich syndrome protein-family verprolin homologous protein 1 (WAVE1)/p21-activated kinase (PAK) phosphorylation and reduced activity of cofilin. These effects of nArgBP2 KD in spines were rescued by inhibiting PAK or activating cofilin combined with sequestration of WAVE. Together, our results suggest that nArgBP2 functions to regulate spine morphogenesis and subsequent spine-synapse formation at glutamatergic synapses. They also raise the possibility that the aberrant regulation of synaptic actin filaments caused by reduced nArgBP2 expression may contribute to the manifestation of the synaptic dysfunction observed in manic/bipolar disorder.

  11. The mevalonate pathway regulates primitive streak formation via protein farnesylation

    PubMed Central

    Okamoto-Uchida, Yoshimi; Yu, Ruoxing; Miyamura, Norio; Arima, Norie; Ishigami-Yuasa, Mari; Kagechika, Hiroyuki; Yoshida, Suguru; Hosoya, Takamitsu; Nawa, Makiko; Kasama, Takeshi; Asaoka, Yoichi; Alois, Reiner Wimmer; Elling, Ulrich; Penninger, Josef M.; Nishina, Sachiko; Azuma, Noriyuki; Nishina, Hiroshi

    2016-01-01

    The primitive streak in peri-implantation embryos forms the mesoderm and endoderm and controls cell differentiation. The metabolic cues regulating primitive streak formation remain largely unknown. Here we utilised a mouse embryonic stem (ES) cell differentiation system and a library of well-characterised drugs to identify these metabolic factors. We found that statins, which inhibit the mevalonate metabolic pathway, suppressed primitive streak formation in vitro and in vivo. Using metabolomics and pharmacologic approaches we identified the downstream signalling pathway of mevalonate and revealed that primitive streak formation requires protein farnesylation but not cholesterol synthesis. A tagging-via-substrate approach revealed that nuclear lamin B1 and small G proteins were farnesylated in embryoid bodies and important for primitive streak gene expression. In conclusion, protein farnesylation driven by the mevalonate pathway is a metabolic cue essential for primitive streak formation. PMID:27883036

  12. Structure of the mitochondrial creatine kinase octamer: high-resolution shadowing and image averaging of single molecules and formation of linear filaments under specific staining conditions

    PubMed Central

    1991-01-01

    The combination of high-resolution tantalum/tungsten (Ta/W) shadowing at very low specimen temperature (-250 degrees C) under ultrahigh vacuum (less than 2 x 10(-9) mbar) with circular harmonic image averaging revealed details on the surface structure of mitochondrial creatine kinase (Mi-CK) molecules with a resolution less than 2.5 nm. Mi-CK octamers exhibit a cross-like surface depression dividing the square shaped projection of 10 x 10 nm into four equally sized subdomains, which correspond to the four dimers forming the octameric Mi-CK molecule. By a combination of positive staining (with uranyl acetate) and heavy metal shadowing, internal structures as well as the surface relief of Mi-CK were visualized at the same time at high resolution. Computational image analysis revealed only a single projection class of molecules, but the ability of Mi-CK to form linear filaments, as well as geometrical considerations concerning the formation of octamers by four equal, asymmetric dimers, suggest the existence of at least two distinct faces on the molecule. By image processing of Mi-CK filaments a side view of the octamer differing from the top-bottom projections of single molecules became evident showing a funnel-like access each form the top and bottom of the octamer connected by a central channel. The general structure of the Mi-CK octamer described here is relevant to the localization of the molecule at the inner-outer mitochondrial contact sites and to the function of Mi-CK as an "energy channeling" molecule. PMID:1702444

  13. The gene for a lectin-like protein is transcriptionally activated during sexual development, but is not essential for fruiting body formation in the filamentous fungus Sordaria macrospora

    PubMed Central

    Nowrousian, Minou; Cebula, Patricia

    2005-01-01

    Background The filamentous fungus Sordaria macrospora forms complex three-dimensional fruiting bodies called perithecia that protect the developing ascospores and ensure their proper discharge. In previous microarray analyses, several genes have been identified that are downregulated in sterile mutants compared to the wild type. Among these genes was tap1 (transcript associated with perithecial development), a gene encoding a putative lectin homolog. Results Analysis of tap1 transcript levels in the wild type under conditions allowing only vegetative growth compared to conditions that lead to fruiting body development showed that tap1 is not only downregulated in developmental mutants but is also upregulated in the wild type during fruiting body development. We have cloned and sequenced a 3.2 kb fragment of genomic DNA containing the tap1 open reading frame and adjoining sequences. The genomic region comprising tap1 is syntenic to its homologous region in the closely related filamentous fungus Neurospora crassa. To determine whether tap1 is involved in fruiting body development in S. macrospora, a knockout construct was generated in which the tap1 open reading frame was replaced by the hygromycin B resistance gene hph under the control of fungal regulatory regions. Transformation of the S. macrospora wild type with this construct resulted in a tap1 deletion strain where tap1 had been replaced by the hph cassette. The knockout strain displayed no phenotypic differences under conditions of vegetative growth and sexual development when compared to the wild type. Double mutants carrying the Δtap1 allele in several developmental mutant backgrounds were phenotypically similar to the corresponding developmental mutant strains. Conclusion The tap1 transcript is strongly upregulated during sexual development in S. macrospora; however, analysis of a tap1 knockout strain shows that tap1 is not essential for fruiting body formation in S. macrospora. PMID:16266439

  14. The gene for a lectin-like protein is transcriptionally activated during sexual development, but is not essential for fruiting body formation in the filamentous fungus Sordaria macrospora.

    PubMed

    Nowrousian, Minou; Cebula, Patricia

    2005-11-03

    The filamentous fungus Sordaria macrospora forms complex three-dimensional fruiting bodies called perithecia that protect the developing ascospores and ensure their proper discharge. In previous microarray analyses, several genes have been identified that are downregulated in sterile mutants compared to the wild type. Among these genes was tap1 (transcript associated with perithecial development), a gene encoding a putative lectin homolog. Analysis of tap1 transcript levels in the wild type under conditions allowing only vegetative growth compared to conditions that lead to fruiting body development showed that tap1 is not only downregulated in developmental mutants but is also upregulated in the wild type during fruiting body development. We have cloned and sequenced a 3.2 kb fragment of genomic DNA containing the tap1 open reading frame and adjoining sequences. The genomic region comprising tap1 is syntenic to its homologous region in the closely related filamentous fungus Neurospora crassa. To determine whether tap1 is involved in fruiting body development in S. macrospora, a knockout construct was generated in which the tap1 open reading frame was replaced by the hygromycin B resistance gene hph under the control of fungal regulatory regions. Transformation of the S. macrospora wild type with this construct resulted in a tap1 deletion strain where tap1 had been replaced by the hph cassette. The knockout strain displayed no phenotypic differences under conditions of vegetative growth and sexual development when compared to the wild type. Double mutants carrying the Deltatap1 allele in several developmental mutant backgrounds were phenotypically similar to the corresponding developmental mutant strains. The tap1 transcript is strongly upregulated during sexual development in S. macrospora; however, analysis of a tap1 knockout strain shows that tap1 is not essential for fruiting body formation in S. macrospora.

  15. Filament turbulence

    NASA Astrophysics Data System (ADS)

    Davidsen, Joern

    2010-03-01

    How much information do you need to distinguish between different mechanisms for spatiotemporal chaos in three-dimensions? In this talk, I will show that the observation of the dynamics on the surface of a medium can be sufficient. Studying mechanisms for filament turbulence in the context of reaction-diffusion media, we found numerically that two major classes of instabilities leave a very different signature on what can be observed on the surface of a three-dimensional medium. These results are of direct relevance in the context of ventricular fibrillation - a turbulent electrical wave activity that destroys the coherent contraction of the ventricular muscle and its main pumping function leading to sudden cardiac death. While it has been proposed that the three-dimensional structure of the heart plays an important role in this type of filament turbulence, only the surface of the heart is currently accessible to experimental observation preventing the study of the full dynamics. Our results suggest that such observations might be sufficient.

  16. N-CADHERIN PRODOMAIN CLEAVAGE REGULATES SYNAPSE FORMATION IN VIVO

    PubMed Central

    Latefi, Nazlie S.; Pedraza, Liliana; Schohl, Anne; Li, Ziwei; Ruthazer, Edward S.

    2009-01-01

    Cadherins are initially synthesized bearing a prodomain that is thought to limit adhesion during early stages of biosynthesis. Functional cadherins lack this prodomain, raising the intriguing possibility that cells may utilize prodomain cleavage as a means to temporally or spatially regulate adhesion after delivery of cadherin to the cell surface. In support of this idea, immunostaining for the prodomain of zebrafish N-cadherin revealed enriched labeling at neuronal surfaces at the soma and along axonal processes. To determine whether post-translational cleavage of the prodomain affects synapse formation, we imaged Rohon-Beard cells in zebrafish embryos expressing GFP-tagged wild-type N-cadherin (NCAD-GFP) or a GFP-tagged N-cadherin mutant expressing an uncleavable prodomain (PRON-GFP) rendering it non-adhesive. NCAD-GFP accumulated at synaptic microdomains in a developmentally regulated manner, and its overexpression transiently accelerated synapse formation. PRON-GFP was much more diffusely distributed along the axon and its overexpression delayed synapse formation. Our results support the notion that N-cadherin serves to stabilize pre- to postsynaptic contacts early in synapse development and suggests that regulated cleavage of the N-cadherin prodomain may be a mechanism by which the kinetics of synaptogenesis are regulated. PMID:19365814

  17. N-cadherin prodomain cleavage regulates synapse formation in vivo.

    PubMed

    Latefi, Nazlie S; Pedraza, Liliana; Schohl, Anne; Li, Ziwei; Ruthazer, Edward S

    2009-07-01

    Cadherins are initially synthesized bearing a prodomain that is thought to limit adhesion during early stages of biosynthesis. Functional cadherins lack this prodomain, raising the intriguing possibility that cells may utilize prodomain cleavage as a means to temporally or spatially regulate adhesion after delivery of cadherin to the cell surface. In support of this idea, immunostaining for the prodomain of zebrafish N-cadherin revealed enriched labeling at neuronal surfaces at the soma and along axonal processes. To determine whether post-translational cleavage of the prodomain affects synapse formation, we imaged Rohon-Beard cells in zebrafish embryos expressing GFP-tagged wild-type N-cadherin (NCAD-GFP) or a GFP-tagged N-cadherin mutant expressing an uncleavable prodomain (PRON-GFP) rendering it nonadhesive. NCAD-GFP accumulated at synaptic microdomains in a developmentally regulated manner, and its overexpression transiently accelerated synapse formation. PRON-GFP was much more diffusely distributed along the axon and its overexpression delayed synapse formation. Our results support the notion that N-cadherin serves to stabilize pre- to postsynaptic contacts early in synapse development and suggests that regulated cleavage of the N-cadherin prodomain may be a mechanism by which the kinetics of synaptogenesis are regulated.

  18. DWARF GALAXY FORMATION WITH H{sub 2}-REGULATED STAR FORMATION

    SciTech Connect

    Kuhlen, Michael; Krumholz, Mark R.; Madau, Piero; Smith, Britton D.; Wise, John

    2012-04-10

    We describe cosmological galaxy formation simulations with the adaptive mesh refinement code Enzo that incorporate a star formation prescription regulated by the local abundance of molecular hydrogen. We show that this H{sub 2}-regulated prescription leads to a suppression of star formation in low-mass halos (M{sub h} {approx}< 10{sup 10} M{sub Sun }) at z > 4, alleviating some of the dwarf galaxy problems faced by theoretical galaxy formation models. H{sub 2} regulation modifies the efficiency of star formation of cold gas directly, rather than indirectly reducing the cold gas content with 'supernova feedback'. We determine the local H{sub 2} abundance in our most refined grid cells (76 proper parsec in size at z = 4) by applying the model of Krumholz, McKee, and Tumlinson, which is based on idealized one-dimensional radiative transfer calculations of H{sub 2} formation-dissociation balance in {approx}100 pc atomic-molecular complexes. Our H{sub 2}-regulated simulations are able to reproduce the empirical (albeit lower z) Kennicutt-Schmidt relation, including the low {Sigma}{sub gas} cutoff due to the transition from atomic to molecular phase and the metallicity dependence thereof, without the use of an explicit density threshold in our star formation prescription. We compare the evolution of the luminosity function, stellar mass density, and star formation rate density from our simulations to recent observational determinations of the same at z = 4-8 and find reasonable agreement between the two.

  19. Micromechanical Thermal Assays of Ca2+-Regulated Thin-Filament Function and Modulation by Hypertrophic Cardiomyopathy Mutants of Human Cardiac Troponin

    PubMed Central

    Brunet, Nicolas M.; Mihajlović, Goran; Aledealat, Khaled; Wang, Fang; Xiong, Peng; von Molnár, Stephan; Chase, P. Bryant

    2012-01-01

    Microfabricated thermoelectric controllers can be employed to investigate mechanisms underlying myosin-driven sliding of Ca2+-regulated actin and disease-associated mutations in myofilament proteins. Specifically, we examined actin filament sliding—with or without human cardiac troponin (Tn) and α-tropomyosin (Tm)—propelled by rabbit skeletal heavy meromyosin, when temperature was varied continuously over a wide range (∼20–63°C). At the upper end of this temperature range, reversible dysregulation of thin filaments occurred at pCa 9 and 5; actomyosin function was unaffected. Tn-Tm enhanced sliding speed at pCa 5 and increased a transition temperature (Tt) between a high activation energy (Ea) but low temperature regime and a low Ea but high temperature regime. This was modulated by factors that alter cross-bridge number and kinetics. Three familial hypertrophic cardiomyopathy (FHC) mutations, cTnI R145G, cTnI K206Q, and cTnT R278C, cause dysregulation at temperatures ∼5–8°C lower; the latter two increased speed at pCa 5 at all temperatures. PMID:22500102

  20. Photospheric flows around a quiescent filament

    NASA Astrophysics Data System (ADS)

    Rondi, S.; Roudier, Th.; Molodij, G.; Bommier, V.; Keil, S.; Sütterlin, P.; Malherbe, J. M.; Meunier, N.; Schmieder, B.; Maloney, P.

    2007-06-01

    Context: The horizontal photospheric flows below and around a filament are one of the components in the formation and evolution of filaments. Few studies exist because they require multiwalength time sequences at high spatial resolution. Aims: Our objective is to measure the horizontal photospheric flows associated with the evolution and eruption of a filament. Methods: We present observations obtained in 2004 during the international JOP 178 campaign which involved eleven instruments both in space and at ground based observatories. We use TRACE WL, DOT and DST observation to derive flow maps which are then coaligned with intensity images and with the vector magnetic field map obtained with THEMIS/MTR. Results: Several supergranulation cells cross the Polarity Inversion Line (PIL) and can transport magnetic flux through the PIL, in particular parasitic polarities. We present a detailed example of the formation of a secondary magnetic dip at the location of a filament footpoint. Large-scale converging flows, which could exist along the filament channel and contribute to its formation, are not observed. Before the filament's eruptive phase, we observe both parasitic and normal polarities being swept by a continuously diverging horizontal flow located in the filament gap. The disappearance of the filament initiates in this gap. Such purely horizontal motions could lead to destabilization of the filament and could trigger the sudden filament disappearance.

  1. Perturbation growth in accreting filaments

    NASA Astrophysics Data System (ADS)

    Clarke, S. D.; Whitworth, A. P.; Hubber, D. A.

    2016-05-01

    We use smoothed particle hydrodynamic simulations to investigate the growth of perturbations in infinitely long filaments as they form and grow by accretion. The growth of these perturbations leads to filament fragmentation and the formation of cores. Most previous work on this subject has been confined to the growth and fragmentation of equilibrium filaments and has found that there exists a preferential fragmentation length-scale which is roughly four times the filament's diameter. Our results show a more complicated dispersion relation with a series of peaks linking perturbation wavelength and growth rate. These are due to gravo-acoustic oscillations along the longitudinal axis during the sub-critical phase of growth. The positions of the peaks in growth rate have a strong dependence on both the mass accretion rate onto the filament and the temperature of the gas. When seeded with a multiwavelength density power spectrum, there exists a clear preferred core separation equal to the largest peak in the dispersion relation. Our results allow one to estimate a minimum age for a filament which is breaking up into regularly spaced fragments, as well as an average accretion rate. We apply the model to observations of filaments in Taurus by Tafalla & Hacar and find accretion rates consistent with those estimated by Palmeirim et al.

  2. Characterizing filaments in regions of high-mass star formation: High-resolution submilimeter imaging of the massive star-forming complex NGC 6334 with ArTéMiS

    NASA Astrophysics Data System (ADS)

    André, Ph.; Revéret, V.; Könyves, V.; Arzoumanian, D.; Tigé, J.; Gallais, P.; Roussel, H.; Le Pennec, J.; Rodriguez, L.; Doumayrou, E.; Dubreuil, D.; Lortholary, M.; Martignac, J.; Talvard, M.; Delisle, C.; Visticot, F.; Dumaye, L.; De Breuck, C.; Shimajiri, Y.; Motte, F.; Bontemps, S.; Hennemann, M.; Zavagno, A.; Russeil, D.; Schneider, N.; Palmeirim, P.; Peretto, N.; Hill, T.; Minier, V.; Roy, A.; Rygl, K. L. J.

    2016-07-01

    Context. Herschel observations of nearby molecular clouds suggest that interstellar filaments and prestellar cores represent two fundamental steps in the star formation process. The observations support a picture of low-mass star formation according to which filaments of ~0.1 pc width form first in the cold interstellar medium, probably as a result of large-scale compression of interstellar matter by supersonic turbulent flows, and then prestellar cores arise from gravitational fragmentation of the densest filaments. Whether this scenario also applies to regions of high-mass star formation is an open question, in part because the resolution of Herschel is insufficient to resolve the inner width of filaments in the nearest regions of massive star formation. Aims: In an effort to characterize the inner width of filaments in high-mass star-forming regions, we imaged the central part of the NGC 6334 complex at a resolution higher by a factor of >3 than Herschel at 350 μm. Methods: We used the large-format bolometer camera ArTéMiS on the APEX telescope and combined the high-resolution ArTéMiS data at 350 μm with Herschel/HOBYS data at 70-500 μm to ensure good sensitivity to a broad range of spatial scales. This allowed us to study the structure of the main narrow filament of the complex with a resolution of 8″ or <0.07 pc at d ~ 1.7 kpc. Results: Our study confirms that this filament is a very dense, massive linear structure with a line mass ranging from ~500 M⊙/pc to ~2000 M⊙/pc over nearly 10 pc. It also demonstrates for the first time that its inner width remains as narrow as W ~ 0.15 ± 0.05 pc all along the filament length, within a factor of <2 of the characteristic 0.1 pc value found with Herschel for lower-mass filaments in the Gould Belt. Conclusions: While it is not completely clear whether the NGC 6334 filament will form massive stars in the future, it is two to three orders of magnitude denser than the majority of filaments observed in Gould Belt

  3. Mechanism of Actin Filament Bundling by Fascin

    SciTech Connect

    Jansen, Silvia; Collins, Agnieszka; Yang, Changsong; Rebowski, Grzegorz; Svitkina, Tatyana; Dominguez, Roberto

    2013-03-07

    Fascin is the main actin filament bundling protein in filopodia. Because of the important role filopodia play in cell migration, fascin is emerging as a major target for cancer drug discovery. However, an understanding of the mechanism of bundle formation by fascin is critically lacking. Fascin consists of four {beta}-trefoil domains. Here, we show that fascin contains two major actin-binding sites, coinciding with regions of high sequence conservation in {beta}-trefoil domains 1 and 3. The site in {beta}-trefoil-1 is located near the binding site of the fascin inhibitor macroketone and comprises residue Ser-39, whose phosphorylation by protein kinase C down-regulates actin bundling and formation of filopodia. The site in {beta}-trefoil-3 is related by pseudo-2-fold symmetry to that in {beta}-trefoil-1. The two sites are {approx}5 nm apart, resulting in a distance between actin filaments in the bundle of {approx}8.1 nm. Residue mutations in both sites disrupt bundle formation in vitro as assessed by co-sedimentation with actin and electron microscopy and severely impair formation of filopodia in cells as determined by rescue experiments in fascin-depleted cells. Mutations of other areas of the fascin surface also affect actin bundling and formation of filopodia albeit to a lesser extent, suggesting that, in addition to the two major actin-binding sites, fascin makes secondary contacts with other filaments in the bundle. In a high resolution crystal structure of fascin, molecules of glycerol and polyethylene glycol are bound in pockets located within the two major actin-binding sites. These molecules could guide the rational design of new anticancer fascin inhibitors.

  4. Carotenoid cation formation and the regulation of photosynthetic light harvesting.

    PubMed

    Holt, Nancy E; Zigmantas, Donatas; Valkunas, Leonas; Li, Xiao-Ping; Niyogi, Krishna K; Fleming, Graham R

    2005-01-21

    Photosynthetic light harvesting in excess light is regulated by a process known as feedback deexcitation. Femtosecond transient absorption measurements on thylakoid membranes show selective formation of a carotenoid radical cation upon excitation of chlorophyll under conditions of maximum, steady-state feedback deexcitation. Studies on transgenic Arabidopsis thaliana plants confirmed that this carotenoid radical cation formation is correlated with feedback deexcitation and requires the presence of zeaxanthin, the specific carotenoid synthesized during high light exposure. These results indicate that energy transfer from chlorophyll molecules to a chlorophyllzeaxanthin heterodimer, which then undergoes charge separation, is the mechanism for excess energy dissipation during feedback deexcitation.

  5. FAM83H and casein kinase I regulate the organization of the keratin cytoskeleton and formation of desmosomes

    PubMed Central

    Kuga, Takahisa; Sasaki, Mitsuho; Mikami, Toshinari; Miake, Yasuo; Adachi, Jun; Shimizu, Maiko; Saito, Youhei; Koura, Minako; Takeda, Yasunori; Matsuda, Junichiro; Tomonaga, Takeshi; Nakayama, Yuji

    2016-01-01

    FAM83H is essential for the formation of dental enamel because a mutation in the FAM83H gene causes amelogenesis imperfecta (AI). We previously reported that the overexpression of FAM83H often occurs and disorganizes the keratin cytoskeleton in colorectal cancer cells. We herein show that FAM83H regulates the organization of the keratin cytoskeleton and maintains the formation of desmosomes in ameloblastoma cells. FAM83H is expressed and localized on keratin filaments in human ameloblastoma cell lines and in mouse ameloblasts and epidermal germinative cells in vivo. FAM83H shows preferential localization to keratin filaments around the nucleus that often extend to cell-cell junctions. Alterations in the function of FAM83H by its overexpression, knockdown, or an AI-causing truncated mutant prevent the proper organization of the keratin cytoskeleton in ameloblastoma cells. Furthermore, the AI-causing mutant prevents desmosomal proteins from being localized to cell-cell junctions. The effects of the AI-causing mutant depend on its binding to and possible inhibition of casein kinase I (CK-1). The suppression of CK-1 by its inhibitor, D4476, disorganizes the keratin cytoskeleton. Our results suggest that AI caused by the FAM83H mutation is mediated by the disorganization of the keratin cytoskeleton and subsequent disruption of desmosomes in ameloblasts. PMID:27222304

  6. High resolution characterization of myosin IIC protein tailpiece and its effect on filament assembly.

    PubMed

    Rosenberg, Masha M; Ronen, Daniel; Lahav, Noa; Nazirov, Elvira; Ravid, Shoshana; Friedler, Assaf

    2013-04-05

    The motor protein nonmuscle myosin II (NMII) must undergo dynamic oligomerization into filaments to perform its cellular functions. A small nonhelical region at the tail of the long coiled-coil region (tailpiece) is a common feature of all dynamically assembling myosin II proteins. This tailpiece is a key regulatory domain affecting NMII filament assembly properties and is subject to phosphorylation in vivo. We previously demonstrated that the positively charged region of the tailpiece binds to assembly-incompetent NMII-C fragments, inducing filament assembly. In the current study, we investigated the molecular mechanisms by which the tailpiece regulates NMII-C self-assembly. Using alanine scan, we found that specific positive and aromatic residues within the positively charged region of the tailpiece are important for inducing NMII-C filament assembly and for filament elongation. Combining peptide arrays with deletion studies allowed us to identify the tailpiece binding sites in the coiled-coil rod. Elucidation of the mechanism by which the tailpiece induces filament assembly permitted us further investigation into the role of tailpiece phosphorylation. Sedimentation and CD spectroscopy identified that phosphorylation of Thr(1957) or Thr(1960) inhibited the ability of the tailpiece to bind the coiled-coil rod and to induce NMII-C filament formation. This study provides molecular insight into the role of specific residues within the NMII-C tailpiece that are responsible for shifting the oligomeric equilibrium of NMII-C toward filament assembly and determining its morphology.

  7. Arabidopsis microtubule-destabilizing protein 25 functions in pollen tube growth by severing actin filaments.

    PubMed

    Qin, Tao; Liu, Xiaomin; Li, Jiejie; Sun, Jingbo; Song, Leina; Mao, Tonglin

    2014-01-01

    The formation of distinct actin filament arrays in the subapical region of pollen tubes is crucial for pollen tube growth. However, the molecular mechanisms underlying the organization and dynamics of the actin filaments in this region remain to be determined. This study shows that Arabidopsis thaliana MICROTUBULE-DESTABILIZING PROTEIN25 (MDP25) has the actin filament-severing activity of an actin binding protein. This protein negatively regulated pollen tube growth by modulating the organization and dynamics of actin filaments in the subapical region of pollen tubes. MDP25 loss of function resulted in enhanced pollen tube elongation and inefficient fertilization. MDP25 bound directly to actin filaments and severed individual actin filaments, in a manner that was dramatically enhanced by Ca(2+), in vitro. Analysis of a mutant that bears a point mutation at the Ca(2+) binding sites demonstrated that the subcellular localization of MDP25 was determined by cytosolic Ca(2+) level in the subapical region of pollen tubes, where MDP25 was disassociated from the plasma membrane and moved into the cytosol. Time-lapse analysis showed that the F-actin-severing frequency significantly decreased and a high density of actin filaments was observed in the subapical region of mdp25-1 pollen tubes. This study reveals a mechanism whereby calcium enhances the actin filament-severing activity of MDP25 in the subapical region of pollen tubes to modulate pollen tube growth.

  8. Regulation of cell proliferation by ERK and signal-dependent nuclear translocation of ERK is dependent on Tm5NM1-containing actin filaments.

    PubMed

    Schevzov, Galina; Kee, Anthony J; Wang, Bin; Sequeira, Vanessa B; Hook, Jeff; Coombes, Jason D; Lucas, Christine A; Stehn, Justine R; Musgrove, Elizabeth A; Cretu, Alexandra; Assoian, Richard; Fath, Thomas; Hanoch, Tamar; Seger, Rony; Pleines, Irina; Kile, Benjamin T; Hardeman, Edna C; Gunning, Peter W

    2015-07-01

    ERK-regulated cell proliferation requires multiple phosphorylation events catalyzed first by MEK and then by casein kinase 2 (CK2), followed by interaction with importin7 and subsequent nuclear translocation of pERK. We report that genetic manipulation of a core component of the actin filaments of cancer cells, the tropomyosin Tm5NM1, regulates the proliferation of normal cells both in vitro and in vivo. Mouse embryo fibroblasts (MEFs) lacking Tm5NM1, which have reduced proliferative capacity, are insensitive to inhibition of ERK by peptide and small-molecule inhibitors, indicating that ERK is unable to regulate proliferation of these knockout (KO) cells. Treatment of wild-type MEFs with a CK2 inhibitor to block phosphorylation of the nuclear translocation signal in pERK resulted in greatly decreased cell proliferation and a significant reduction in the nuclear translocation of pERK. In contrast, Tm5NM1 KO MEFs, which show reduced nuclear translocation of pERK, were unaffected by inhibition of CK2. This suggested that it is nuclear translocation of CK2-phosphorylated pERK that regulates cell proliferation and this capacity is absent in Tm5NM1 KO cells. Proximity ligation assays confirmed a growth factor-stimulated interaction of pERK with Tm5NM1 and that the interaction of pERK with importin7 is greatly reduced in the Tm5NM1 KO cells.

  9. Regulation of cell proliferation by ERK and signal-dependent nuclear translocation of ERK is dependent on Tm5NM1-containing actin filaments

    PubMed Central

    Schevzov, Galina; Kee, Anthony J.; Wang, Bin; Sequeira, Vanessa B.; Hook, Jeff; Coombes, Jason D.; Lucas, Christine A.; Stehn, Justine R.; Musgrove, Elizabeth A.; Cretu, Alexandra; Assoian, Richard; Fath, Thomas; Hanoch, Tamar; Seger, Rony; Pleines, Irina; Kile, Benjamin T.; Hardeman, Edna C.; Gunning, Peter W.

    2015-01-01

    ERK-regulated cell proliferation requires multiple phosphorylation events catalyzed first by MEK and then by casein kinase 2 (CK2), followed by interaction with importin7 and subsequent nuclear translocation of pERK. We report that genetic manipulation of a core component of the actin filaments of cancer cells, the tropomyosin Tm5NM1, regulates the proliferation of normal cells both in vitro and in vivo. Mouse embryo fibroblasts (MEFs) lacking Tm5NM1, which have reduced proliferative capacity, are insensitive to inhibition of ERK by peptide and small-molecule inhibitors, indicating that ERK is unable to regulate proliferation of these knockout (KO) cells. Treatment of wild-type MEFs with a CK2 inhibitor to block phosphorylation of the nuclear translocation signal in pERK resulted in greatly decreased cell proliferation and a significant reduction in the nuclear translocation of pERK. In contrast, Tm5NM1 KO MEFs, which show reduced nuclear translocation of pERK, were unaffected by inhibition of CK2. This suggested that it is nuclear translocation of CK2-phosphorylated pERK that regulates cell proliferation and this capacity is absent in Tm5NM1 KO cells. Proximity ligation assays confirmed a growth factor–stimulated interaction of pERK with Tm5NM1 and that the interaction of pERK with importin7 is greatly reduced in the Tm5NM1 KO cells. PMID:25971798

  10. The Saccharomyces cerevisiae 14-3-3 protein Bmh2 is required for regulation of the phosphorylation status of Fin1, a novel intermediate filament protein.

    PubMed Central

    Mayordomo, Isabel; Sanz, Pascual

    2002-01-01

    In order to identify proteins that interact with Bmh2, a yeast member of the 14-3-3 protein family, we performed a two-hybrid screening using LexA-Bmh2 as bait. We identified Fin1, a novel intermediate filament protein, as the protein that showed the highest degree of interaction. We also identified components of the vesicular transport machinery such as Gic2 and Msb3, proteins involved in transcriptional regulation such as Mbf1, Gcr2 and Reg2, and a variety of other different proteins (Ppt1, Lre1, Rps0A and Ylr177w). We studied the interaction between Bmh2 and Fin1 in more detail and found that Bmh2 only interacted with phosphorylated forms of Fin1. In addition, we showed that Glc7, the catalytic subunit of the protein phosphatase 1 complex, was also able to interact with Fin1. PMID:11931638

  11. TGF-β Signaling Regulates Cementum Formation through Osterix Expression

    PubMed Central

    Choi, Hwajung; Ahn, Yu-Hyun; Kim, Tak-Heun; Bae, Cheol-Hyeon; Lee, Jeong-Chae; You, Hyung-Keun; Cho, Eui-Sic

    2016-01-01

    TGF-β/BMPs have widely recognized roles in mammalian development, including in bone and tooth formation. To define the functional relevance of the autonomous requirement for TGF-β signaling in mouse tooth development, we analyzed osteocalcin-Cre mediated Tgfbr2 (OCCreTgfbr2fl/fl) conditional knockout mice, which lacks functional TGF-β receptor II (TβRII) in differentiating cementoblasts and cementocytes. Strikingly, OCCreTgfbr2fl/fl mutant mice exhibited a sharp reduction in cellular cementum mass with reduced matrix secretion and mineral apposition rates. To explore the molecular mechanisms underlying the roles of TGF-β signaling through TβRII in cementogenesis, we established a mouse cementoblast model with decreased TβRII expression using OCCM-30 cells. Interestingly, the expression of osterix (Osx), one of the major regulators of cellular cementum formation, was largely decreased in OCCM-30 cells lacking TβRII. Consequently, in those cells, functional ALP activity and the expression of genes associated with cementogenesis were reduced and the cells were partially rescued by Osx transduction. We also found that TGF-β signaling directly regulates Osx expression through a Smad-dependent pathway. These findings strongly suggest that TGF-β signaling plays a major role as one of the upstream regulators of Osx in cementoblast differentiation and cementum formation. PMID:27180803

  12. Formation and Regulation of Adaptive Response in Nematode Caenorhabditis elegans

    PubMed Central

    Zhao, Y.-L.; Wang, D.-Y.

    2012-01-01

    All organisms respond to environmental stresses (e.g., heavy metal, heat, UV irradiation, hyperoxia, food limitation, etc.) with coordinated adjustments in order to deal with the consequences and/or injuries caused by the severe stress. The nematode Caenorhabditis elegans often exerts adaptive responses if preconditioned with low concentrations of agents or stressor. In C. elegans, three types of adaptive responses can be formed: hormesis, cross-adaptation, and dietary restriction. Several factors influence the formation of adaptive responses in nematodes, and some mechanisms can explain their response formation. In particular, antioxidation system, heat-shock proteins, metallothioneins, glutathione, signaling transduction, and metabolic signals may play important roles in regulating the formation of adaptive responses. In this paper, we summarize the published evidence demonstrating that several types of adaptive responses have converged in C. elegans and discussed some possible alternative theories explaining the adaptive response control. PMID:22997543

  13. The Link between Turbulence, Magnetic Fields, Filaments, and Star Formation in the Central Molecular Zone Cloud G0.253+0.016

    NASA Astrophysics Data System (ADS)

    Federrath, C.; Rathborne, J. M.; Longmore, S. N.; Kruijssen, J. M. D.; Bally, J.; Contreras, Y.; Crocker, R. M.; Garay, G.; Jackson, J. M.; Testi, L.; Walsh, A. J.

    2016-12-01

    Star formation is primarily controlled by the interplay between gravity, turbulence, and magnetic fields. However, the turbulence and magnetic fields in molecular clouds near the Galactic center may differ substantially compared to spiral-arm clouds. Here we determine the physical parameters of the central molecular zone (CMZ) cloud G0.253+0.016, its turbulence, magnetic field, and filamentary structure. Using column density maps based on dust-continuum emission observations with ALMA+Herschel, we identify filaments and show that at least one dense core is located along them. We measure the filament width {W}{fil}=0.17+/- 0.08 {pc} and the sonic scale {λ }{sonic}=0.15+/- 0.11 {pc} of the turbulence, and find {W}{fil}≈ {λ }{sonic}. A strong velocity gradient is seen in the HNCO intensity-weighted velocity maps obtained with ALMA+Mopra. The gradient is likely caused by large-scale shearing of G0.253+0.016, producing a wide double-peaked velocity probability distribution function (PDF). After subtracting the gradient to isolate the turbulent motions, we find a nearly Gaussian velocity PDF typical for turbulence. We measure the total and turbulent velocity dispersion, 8.8+/- 0.2 {km} {{{s}}}-1 and 3.9+/- 0.1 {km} {{{s}}}-1, respectively. Using magnetohydrodynamical turbulence simulations, we find that G0.253+0.016's turbulent magnetic field {B}{turb}=130+/- 50 μ {{G}} is only ≲ 1/10 of the ordered field component. Combining these measurements, we reconstruct the dominant turbulence driving mode in G0.253+0.016 and find a driving parameter of b=0.22+/- 0.12, indicating solenoidal (divergence-free) driving. We compare this to spiral-arm clouds, which typically have a significant compressive (curl-free) driving component (b\\gt 0.4). Motivated by previous reports of strong shearing motions in the CMZ, we speculate that shear causes the solenoidal driving in G0.253+0.016 and show that this reduces the star-formation rate by a factor of 6.9 compared to typical nearby

  14. Confronting the outflow-regulated cluster formation model with observations

    SciTech Connect

    Nakamura, Fumitaka; Li, Zhi-Yun E-mail: zl4h@virginia.edu

    2014-03-10

    Protostellar outflows have been shown theoretically to be capable of maintaining supersonic turbulence in cluster-forming clumps and keeping the star formation rate per free-fall time as low as a few percent. We aim to test two basic predictions of this outflow-regulated cluster formation model, namely, (1) the clump should be close to virial equilibrium and (2) the turbulence dissipation rate should be balanced by the outflow momentum injection rate, using recent outflow surveys toward eight nearby cluster-forming clumps (B59, L1551, L1641N, Serpens Main Cloud, Serpens South, ρ Oph, IC 348, and NGC 1333). We find, for almost all sources, that the clumps are close to virial equilibrium and the outflow momentum injection rate exceeds the turbulence momentum dissipation rate. In addition, the outflow kinetic energy is significantly smaller than the clump gravitational energy for intermediate and massive clumps with M {sub cl} ≳ a few × 10{sup 2} M {sub ☉}, suggesting that the outflow feedback is not enough to disperse the clump as a whole. The number of observed protostars also indicates that the star formation rate per free-fall time is as small as a few percent for all clumps. These observationally based results strengthen the case for outflow-regulated cluster formation.

  15. The Many Fates of Retracting Newtonian Filaments

    NASA Astrophysics Data System (ADS)

    Anthony, Christopher; Thete, Sumeet; Harris, Michael; Basaran, Osman

    2016-11-01

    The retraction of Newtonian filaments plays a central role in applications as diverse as inkjet printing and atomization where formation of satellite droplets is undesirable. In order to avoid satellite drop production, filaments formed after drop, jet, or sheet breakup should contract to spheres without undergoing further pinch-off. Therefore, it is important to understand all of the dynamical responses that can arise during filament recoil. To accomplish this goal, we use high accuracy simulations to analyze the retraction of Newtonian filaments in a passive ambient fluid. Previously, Notz and Basaran described the fate of low-viscosity filaments. More recent works by Hoepffner and Pare on intermediate viscosity filaments and by Lohse et al. on high viscosity filaments have greatly enhanced our understanding of filament recoil. Unfortunately, taking all of these works in aggregate does not provide a comprehensive picture of filament dynamics. Here, we overcome the deficiencies of these earlier studies to provide a comprehensive analysis of filament recoil and arrive at a complete phase diagram of the system response. While doing so, we also uncover a new mode of filament breakup that has been missed by earlier investigators.

  16. Contributions of EspA filaments and curli fimbriae in cellular adherence and biofilm formation of enterohemorrhagic Escherichia coli O157:H7

    USDA-ARS?s Scientific Manuscript database

    In Escherichia coli O157:H7 (O157), the filamentous structure of the type III secretion system is produced from the polymerization of the EspA protein. EspA filaments are essential for O157 adherence to epithelial cells. In previous studies, we demonstrated that O157 hha deletion mutants showed incr...

  17. GMC Collisions as Triggers of Star Formation. III. Density and Magnetically Regulated Star Formation

    NASA Astrophysics Data System (ADS)

    Wu, Benjamin; Tan, Jonathan C.; Christie, Duncan; Nakamura, Fumitaka; Van Loo, Sven; Collins, David

    2017-06-01

    We study giant molecular cloud (GMC) collisions and their ability to trigger star cluster formation. We further develop our three-dimensional magnetized, turbulent, colliding GMC simulations by implementing star formation subgrid models. Two such models are explored: (1) “Density-Regulated,” i.e., fixed efficiency per free-fall time above a set density threshold and (2) “Magnetically Regulated,” i.e., fixed efficiency per free-fall time in regions that are magnetically supercritical. Variations of parameters associated with these models are also explored. In the non-colliding simulations, the overall level of star formation is sensitive to model parameter choices that relate to effective density thresholds. In the GMC collision simulations, the final star formation rates and efficiencies are relatively independent of these parameters. Between the non-colliding and colliding cases, we compare the morphologies of the resulting star clusters, properties of star-forming gas, time evolution of the star formation rate (SFR), spatial clustering of the stars, and resulting kinematics of the stars in comparison to the natal gas. We find that typical collisions, by creating larger amounts of dense gas, trigger earlier and enhanced star formation, resulting in 10 times higher SFRs and efficiencies. The star clusters formed from GMC collisions show greater spatial substructure and more disturbed kinematics.

  18. Pseudomonas fluorescens Filamentous Hemagglutinin, an Iron-Regulated Protein, Is an Important Virulence Factor that Modulates Bacterial Pathogenicity

    PubMed Central

    Sun, Yuan-Yuan; Chi, Heng; Sun, Li

    2016-01-01

    Pseudomonas fluorescens is a common bacterial pathogen to a wide range of aquaculture animals including various species of fish. In this study, we employed proteomic analysis and identified filamentous hemagglutinin (FHA) as an iron-responsive protein secreted by TSS, a pathogenic P. fluorescens isolate. In vitro study showed that compared to the wild type, the fha mutant TSSfha (i) exhibited a largely similar vegetative growth profile but significantly retarded in the ability of biofilm growth and producing extracellular matrix, (ii) displayed no apparent flagella and motility, (iii) was defective in the attachment to host cells and unable to form self-aggregation, (iv) displayed markedly reduced capacity of hemagglutination and surviving in host serum. In vivo infection analysis revealed that TSSfha was significantly attenuated in the ability of dissemination in fish tissues and inducing host mortality, and that antibody blocking of the natural FHA produced by the wild type TSS impaired the infectivity of the pathogen. Furthermore, when introduced into turbot as a subunit vaccine, recombinant FHA elicited a significant protection against lethal TSS challenge. Taken together, these results indicate for the first time that P. fluorescens FHA is a key virulence factor essential to multiple biological processes associated with pathogenicity. PMID:27602029

  19. Role of filamentous phage SW1 in regulating the lateral flagella of Shewanella piezotolerans strain WP3 at low temperatures.

    PubMed

    Jian, Huahua; Xiao, Xiang; Wang, Fengping

    2013-11-01

    Low-temperature ecosystems represent the largest biosphere on Earth, and yet our understanding of the roles of bacteriophages in these systems is limited. Here, the influence of the cold-active filamentous phage SW1 on the phenotype and gene transcription of its host, Shewanella piezotolerans WP3 (WP3), was investigated by construction of a phage-free strain (WP3ΔSW1), which was compared with the wild-type strain. The expression of 49 genes, including 16 lateral flagellar genes, was found to be significantly influenced by SW1 at 4°C, as demonstrated by comparative whole-genome microarray analysis. WP3ΔSW1 was shown to have a higher production of lateral flagella than WP3 and enhanced swarming motility when cultivated on solid agar plates. Besides, SW1 has a remarkable impact on the expression of a variety of host genes in liquid culture, particularly the genes related to the membrane and to the production of lateral flagella. These results suggest that the deep-sea bacterium WP3 might balance the high-energy demands of phage maintenance and swarming motility at low temperatures. The phage SW1 is shown to have a significant influence on the swarming ability of the host and thus may play an important role in adjusting the fitness of the cells in the deep-sea environment.

  20. Role of Filamentous Phage SW1 in Regulating the Lateral Flagella of Shewanella piezotolerans Strain WP3 at Low Temperatures

    PubMed Central

    Jian, Huahua; Xiao, Xiang

    2013-01-01

    Low-temperature ecosystems represent the largest biosphere on Earth, and yet our understanding of the roles of bacteriophages in these systems is limited. Here, the influence of the cold-active filamentous phage SW1 on the phenotype and gene transcription of its host, Shewanella piezotolerans WP3 (WP3), was investigated by construction of a phage-free strain (WP3ΔSW1), which was compared with the wild-type strain. The expression of 49 genes, including 16 lateral flagellar genes, was found to be significantly influenced by SW1 at 4°C, as demonstrated by comparative whole-genome microarray analysis. WP3ΔSW1 was shown to have a higher production of lateral flagella than WP3 and enhanced swarming motility when cultivated on solid agar plates. Besides, SW1 has a remarkable impact on the expression of a variety of host genes in liquid culture, particularly the genes related to the membrane and to the production of lateral flagella. These results suggest that the deep-sea bacterium WP3 might balance the high-energy demands of phage maintenance and swarming motility at low temperatures. The phage SW1 is shown to have a significant influence on the swarming ability of the host and thus may play an important role in adjusting the fitness of the cells in the deep-sea environment. PMID:24038680

  1. Pseudomonas fluorescens Filamentous Hemagglutinin, an Iron-Regulated Protein, Is an Important Virulence Factor that Modulates Bacterial Pathogenicity.

    PubMed

    Sun, Yuan-Yuan; Chi, Heng; Sun, Li

    2016-01-01

    Pseudomonas fluorescens is a common bacterial pathogen to a wide range of aquaculture animals including various species of fish. In this study, we employed proteomic analysis and identified filamentous hemagglutinin (FHA) as an iron-responsive protein secreted by TSS, a pathogenic P. fluorescens isolate. In vitro study showed that compared to the wild type, the fha mutant TSSfha (i) exhibited a largely similar vegetative growth profile but significantly retarded in the ability of biofilm growth and producing extracellular matrix, (ii) displayed no apparent flagella and motility, (iii) was defective in the attachment to host cells and unable to form self-aggregation, (iv) displayed markedly reduced capacity of hemagglutination and surviving in host serum. In vivo infection analysis revealed that TSSfha was significantly attenuated in the ability of dissemination in fish tissues and inducing host mortality, and that antibody blocking of the natural FHA produced by the wild type TSS impaired the infectivity of the pathogen. Furthermore, when introduced into turbot as a subunit vaccine, recombinant FHA elicited a significant protection against lethal TSS challenge. Taken together, these results indicate for the first time that P. fluorescens FHA is a key virulence factor essential to multiple biological processes associated with pathogenicity.

  2. How stellar feedback simultaneously regulates star formation and drives outflows

    NASA Astrophysics Data System (ADS)

    Hayward, Christopher C.; Hopkins, Philip F.

    2017-02-01

    We present an analytic model for how momentum deposition from stellar feedback simultaneously regulates star formation and drives outflows in a turbulent interstellar medium (ISM). Because the ISM is turbulent, a given patch of ISM exhibits sub-patches with a range of surface densities. The high-density patches are 'pushed' by feedback, thereby driving turbulence and self-regulating local star formation. Sufficiently low-density patches, however, are accelerated to above the escape velocity before the region can self-adjust and are thus vented as outflows. When the gas fraction is ≳ 0.3, the ratio of the turbulent velocity dispersion to the circular velocity is sufficiently high that at any given time, of the order of half of the ISM has surface density less than the critical value and thus can be blown out on a dynamical time. The resulting outflows have a mass-loading factor (η ≡ dot{M}_{out}/M_{star }) that is inversely proportional to the gas fraction times the circular velocity. At low gas fractions, the star formation rate needed for local self-regulation, and corresponding turbulent Mach number, declines rapidly; the ISM is 'smoother', and it is actually more difficult to drive winds with large mass-loading factors. Crucially, our model predicts that stellar-feedback-driven outflows should be suppressed at z ≲ 1 in M⋆ ≳ 1010 M⊙ galaxies. This mechanism allows massive galaxies to exhibit violent outflows at high redshifts and then 'shut down' those outflows at late times, thereby enabling the formation of a smooth, extended thin stellar disc. We provide simple fitting functions for η that should be useful for sub-resolution and semi-analytic models.

  3. Specific posttranslational modification regulates early events in mammary carcinoma formation.

    PubMed

    Guo, Hua-Bei; Johnson, Heather; Randolph, Matthew; Nagy, Tamas; Blalock, Ryan; Pierce, Michael

    2010-12-07

    The expression of an enzyme, GnT-V, that catalyzes a specific posttranslational modification of a family of glycoproteins, namely a branched N-glycan, is transcriptionally up-regulated during breast carcinoma oncogenesis. To determine the molecular basis of how early events in breast carcinoma formation are regulated by GnT-V, we studied both the early stages of mammary tumor formation by using 3D cell culture and a her-2 transgenic mouse mammary tumor model. Overexpression of GnT-V in MCF-10A mammary epithelial cells in 3D culture disrupted acinar morphogenesis with impaired hollow lumen formation, an early characteristic of mammary neoplastic transformation. The disrupted acinar morphogenesis of mammary tumor cells in 3D culture caused by her-2 expression was reversed in tumors that lacked GnT-V expression. Moreover, her-2-induced mammary tumor onset was significantly delayed in the GnT-V null tumors, evidence that the lack of the posttranslational modification catalyzed by GnT-V attenuated tumor formation. Inhibited activation of both PKB and ERK signaling pathways was observed in GnT-V null tumor cells. The proportion of tumor-initiating cells (TICs) in the mammary tumors from GnT-V null mice was significantly reduced compared with controls, and GnT-V null TICs displayed a reduced ability to form secondary tumors in NOD/SCID mice. These results demonstrate that GnT-V expression and its branched glycan products effectively modulate her-2-mediated signaling pathways that, in turn, regulate the relative proportion of tumor initiating cells and the latency of her-2-driven tumor onset.

  4. Bordetella pertussis infection of human respiratory epithelial cells up-regulates intercellular adhesion molecule-1 expression: role of filamentous hemagglutinin and pertussis toxin.

    PubMed

    Ishibashi, Yoshio; Nishikawa, Akemi

    2002-09-01

    Adhesion molecules on respiratory epithelial cells play a critical role in inflammatory cell recruitment and accumulation at sites of inflammation. Bordetella pertussis colonizes the human respiratory tract by infecting epithelial cells, leading to an inflammatory response. In this study, the role of bacterial factors in the expression of intercellular adhesion molecule-1 (ICAM-1) on human respiratory epithelial cells was investigated in response to B. pertussis. Flow cytometry and real time RT-PCR analysis showed that BEAS-2B human bronchial epithelial cells expressed increased levels of ICAM-1 mRNA and surface protein in response to B. pertussis infection. Filamentous hemagglutinin (FHA) played a role in this response because of the impaired capability of a FHA-deficient isogenic strain. A mutant strain in which an Arg-Gly-Asp (RGD) site of FHA had been changed to Arg-Ala-Asp had diminished ability to up-regulate ICAM-1 expression. RGD sequence-associated up-regulation of ICAM-1 expression was also observed in primary normal human bronchial epithelial cells. Pretreatment of cells with integrin antagonists such as RGD-containing peptide and antibody against very late antigen-5 (VLA-5) inhibited the up-regulation of ICAM-1 expression, suggesting the participation of VLA-5 integrin in this response. Pertussis toxin (PT) prevented the up-regulation of ICAM-1 expression because a PT-deficient mutant strain induced higher levels of ICAM-1 mRNA and surface protein than the parental strain. Consistent with this, purified PT suppressed the up-regulation of epithelial ICAM-1 expression. These findings demonstrate that B. pertussis FHA up-regulates ICAM-1 expression on respiratory epithelial cells through interaction of its RGD site with host cell VLA-5 integrin, and that PT impairs this response.

  5. Radial Infall onto a Massive Molecular Filament

    NASA Astrophysics Data System (ADS)

    Battersby, Cara

    2015-08-01

    The newly discovered massive molecular filament G32.02+0.05 (~70 pc long, 10^5 Msun) has been shaped and compressed by older generations of massive stars. The similarity of this filament in physical structure (density profile, temperature) to much smaller star-forming filaments, suggests that the mechanism to form such filaments is a universal process. The densest portion of the filament, apparent as an Infrared Dark Cloud (IRDC) appears to have been compressed on both sides by a pair of bubbles and now shows a range of massive star formation signatures throughout. We investigate the kinematics in this massive star-forming filament and find velocity coherence along the filament and global blue asymmetric line profiles. These line asymmetries are interpreted as a signature of large-scale radial collapse. Using line asymmetries observed with optically thick HCO+ (1-0) and optically thin H13CO+ (1-0) across a range of massive star forming regions in the filament, we estimate the global radial infall rate of the filament to range from a few 100 to a few 1000 Msun/Myr/pc^2. At its current infall rate the densest portions of the cloud will more than double their current mass within a Myr. The discovery of such a reservoir of infalling gas has important ramifications on the course of star formation in the cloud and on our overall understanding of star cluster formation.

  6. Autophagy-Associated Protein SmATG12 Is Required for Fruiting-Body Formation in the Filamentous Ascomycete Sordaria macrospora

    PubMed Central

    Werner, Antonia; Herzog, Britta; Frey, Stefan; Pöggeler, Stefanie

    2016-01-01

    In filamentous fungi, autophagy functions as a catabolic mechanism to overcome starvation and to control diverse developmental processes under normal nutritional conditions. Autophagy involves the formation of double-membrane vesicles, termed autophagosomes that engulf cellular components and bring about their degradation via fusion with vacuoles. Two ubiquitin-like (UBL) conjugation systems are essential for the expansion of the autophagosomal membrane: the UBL protein ATG8 is conjugated to the lipid phosphatidylethanolamine and the UBL protein ATG12 is coupled to ATG5. We recently showed that in the homothallic ascomycete Sordaria macrospora autophagy-related genes encoding components of the conjugation systems are required for fruiting-body development and/or are essential for viability. In the present work, we cloned and characterized the S. macrospora (Sm)atg12 gene. Two-hybrid analysis revealed that SmATG12 can interact with SmATG7 and SmATG3. To examine its role in S. macrospora, we replaced the open reading frame of Smatg12 with a hygromycin resistance cassette and generated a homokaryotic ΔSmatg12 knockout strain, which displayed slower vegetative growth under nutrient starvation conditions and was unable to form fruiting bodies. In the hyphae of S. macrospora EGFP-labeled SmATG12 was detected in the cytoplasm and as punctate structures presumed to be phagophores or phagophore assembly sites. Delivery of EGFP-labelled SmATG8 to the vacuole was entirely dependent on SmATG12. PMID:27309377

  7. Autophagy-Associated Protein SmATG12 Is Required for Fruiting-Body Formation in the Filamentous Ascomycete Sordaria macrospora.

    PubMed

    Werner, Antonia; Herzog, Britta; Frey, Stefan; Pöggeler, Stefanie

    2016-01-01

    In filamentous fungi, autophagy functions as a catabolic mechanism to overcome starvation and to control diverse developmental processes under normal nutritional conditions. Autophagy involves the formation of double-membrane vesicles, termed autophagosomes that engulf cellular components and bring about their degradation via fusion with vacuoles. Two ubiquitin-like (UBL) conjugation systems are essential for the expansion of the autophagosomal membrane: the UBL protein ATG8 is conjugated to the lipid phosphatidylethanolamine and the UBL protein ATG12 is coupled to ATG5. We recently showed that in the homothallic ascomycete Sordaria macrospora autophagy-related genes encoding components of the conjugation systems are required for fruiting-body development and/or are essential for viability. In the present work, we cloned and characterized the S. macrospora (Sm)atg12 gene. Two-hybrid analysis revealed that SmATG12 can interact with SmATG7 and SmATG3. To examine its role in S. macrospora, we replaced the open reading frame of Smatg12 with a hygromycin resistance cassette and generated a homokaryotic ΔSmatg12 knockout strain, which displayed slower vegetative growth under nutrient starvation conditions and was unable to form fruiting bodies. In the hyphae of S. macrospora EGFP-labeled SmATG12 was detected in the cytoplasm and as punctate structures presumed to be phagophores or phagophore assembly sites. Delivery of EGFP-labelled SmATG8 to the vacuole was entirely dependent on SmATG12.

  8. Vesicle Size Regulates Nanotube Formation in the Cell.

    PubMed

    Su, Qian Peter; Du, Wanqing; Ji, Qinghua; Xue, Boxin; Jiang, Dong; Zhu, Yueyao; Lou, Jizhong; Yu, Li; Sun, Yujie

    2016-04-07

    Intracellular membrane nanotube formation and its dynamics play important roles for cargo transportation and organelle biogenesis. Regarding the regulation mechanisms, while much attention has been paid on the lipid composition and its associated protein molecules, effects of the vesicle size has not been studied in the cell. Giant unilamellar vesicles (GUVs) are often used for in vitro membrane deformation studies, but they are much larger than most intracellular vesicles and the in vitro studies also lack physiological relevance. Here, we use lysosomes and autolysosomes, whose sizes range between 100 nm and 1 μm, as model systems to study the size effects on nanotube formation both in vivo and in vitro. Single molecule observations indicate that driven by kinesin motors, small vesicles (100-200 nm) are mainly transported along the tracks while a remarkable portion of large vesicles (500-1000 nm) form nanotubes. This size effect is further confirmed by in vitro reconstitution assays on liposomes and purified lysosomes and autolysosomes. We also apply Atomic Force Microscopy (AFM) to measure the initiation force for nanotube formation. These results suggest that the size-dependence may be one of the mechanisms for cells to regulate cellular processes involving membrane-deformation, such as the timing of tubulation-mediated vesicle recycling.

  9. Filamentous Fungi.

    PubMed

    Powers-Fletcher, Margaret V; Kendall, Brian A; Griffin, Allen T; Hanson, Kimberly E

    2016-06-01

    Filamentous mycoses are often associated with significant morbidity and mortality. Prompt diagnosis and aggressive treatment are essential for good clinical outcomes in immunocompromised patients. The host immune response plays an essential role in determining the course of exposure to potential fungal pathogens. Depending on the effectiveness of immune response and the burden of organism exposure, fungi can either be cleared or infection can occur and progress to a potentially fatal invasive disease. Nonspecific cellular immunity (i.e., neutrophils, natural killer [NK] cells, and macrophages) combined with T-cell responses are the main immunologic mechanisms of protection. The most common potential mold pathogens include certain hyaline hyphomycetes, endemic fungi, the Mucorales, and some dematiaceous fungi. Laboratory diagnostics aimed at detecting and differentiating these organisms are crucial to helping clinicians make informed decisions about treatment. The purpose of this chapter is to provide an overview of the medically important fungal pathogens, as well as to discuss the patient characteristics, antifungal-therapy considerations, and laboratory tests used in current clinical practice for the immunocompromised host.

  10. Nonlinear evolution of three-dimensional instabilities of thin and thick electron scale current sheets: Plasmoid formation and current filamentation

    SciTech Connect

    Jain, Neeraj; Büchner, Jörg

    2014-07-15

    Nonlinear evolution of three dimensional electron shear flow instabilities of an electron current sheet (ECS) is studied using electron-magnetohydrodynamic simulations. The dependence of the evolution on current sheet thickness is examined. For thin current sheets (half thickness =d{sub e}=c/ω{sub pe}), tearing mode instability dominates. In its nonlinear evolution, it leads to the formation of oblique current channels. Magnetic field lines form 3-D magnetic spirals. Even in the absence of initial guide field, the out-of-reconnection-plane magnetic field generated by the tearing instability itself may play the role of guide field in the growth of secondary finite-guide-field instabilities. For thicker current sheets (half thickness ∼5 d{sub e}), both tearing and non-tearing modes grow. Due to the non-tearing mode, current sheet becomes corrugated in the beginning of the evolution. In this case, tearing mode lets the magnetic field reconnect in the corrugated ECS. Later thick ECS develops filamentary structures and turbulence in which reconnection occurs. This evolution of thick ECS provides an example of reconnection in self-generated turbulence. The power spectra for both the thin and thick current sheets are anisotropic with respect to the electron flow direction. The cascade towards shorter scales occurs preferentially in the direction perpendicular to the electron flow.

  11. Homeostatic regulation of meiotic DSB formation by ATM/ATR

    SciTech Connect

    Cooper, Tim J.; Wardell, Kayleigh; Garcia, Valerie; Neale, Matthew J.

    2014-11-15

    Ataxia–telangiectasia mutated (ATM) and RAD3-related (ATR) are widely known as being central players in the mitotic DNA damage response (DDR), mounting responses to DNA double-strand breaks (DSBs) and single-stranded DNA (ssDNA) respectively. The DDR signalling cascade couples cell cycle control to damage-sensing and repair processes in order to prevent untimely cell cycle progression while damage still persists [1]. Both ATM/ATR are, however, also emerging as essential factors in the process of meiosis; a specialised cell cycle programme responsible for the formation of haploid gametes via two sequential nuclear divisions. Central to achieving accurate meiotic chromosome segregation is the introduction of numerous DSBs spread across the genome by the evolutionarily conserved enzyme, Spo11. This review seeks to explore and address how cells utilise ATM/ATR pathways to regulate Spo11-DSB formation, establish DSB homeostasis and ensure meiosis is completed unperturbed.

  12. How Stellar Feedback Simultaneously Regulates Star Formation and Drives Outflows

    NASA Astrophysics Data System (ADS)

    Hayward, Christopher C.

    2017-07-01

    Stellar-feedback-driven outflows are thought to be a crucial process in galaxy formation, but our physical understanding of them is still rudimentary. I will present an analytical model for how stellar feedback simultaneously regulates star formation and drives outflows. One interesting prediction of the model is that although stellar feedback can efficiently generate outflows in all galaxies at high redshift, it is unable to drive outflows in massive galaxies below z 1. This prediction agrees with the results of state-of-the-art cosmological zoom simulations that include explicit stellar feedback. This model, combined with the aforementioned simulations, suggests that high-redshift galaxies are highly turbulent, disordered and characterized by strong starbursts and subsequent violent outflows, and the suppression of outflows in massive galaxies at z 1 is crucial for the emergence of well-ordered, steadily star-forming disk galaxies.

  13. Homeostatic regulation of meiotic DSB formation by ATM/ATR.

    PubMed

    Cooper, Tim J; Wardell, Kayleigh; Garcia, Valerie; Neale, Matthew J

    2014-11-15

    Ataxia-telangiectasia mutated (ATM) and RAD3-related (ATR) are widely known as being central players in the mitotic DNA damage response (DDR), mounting responses to DNA double-strand breaks (DSBs) and single-stranded DNA (ssDNA) respectively. The DDR signalling cascade couples cell cycle control to damage-sensing and repair processes in order to prevent untimely cell cycle progression while damage still persists [1]. Both ATM/ATR are, however, also emerging as essential factors in the process of meiosis; a specialised cell cycle programme responsible for the formation of haploid gametes via two sequential nuclear divisions. Central to achieving accurate meiotic chromosome segregation is the introduction of numerous DSBs spread across the genome by the evolutionarily conserved enzyme, Spo11. This review seeks to explore and address how cells utilise ATM/ATR pathways to regulate Spo11-DSB formation, establish DSB homeostasis and ensure meiosis is completed unperturbed.

  14. HDAC2 negatively regulates memory formation and synaptic plasticity

    PubMed Central

    Guan, Ji-Song; Haggarty, Stephen J.; Giacometti, Emanuela; Dannenberg, Jan-Hermen; Joseph, Nadine; Gao, Jun; Nieland, Thomas J.F.; Zhou, Ying; Wang, Xinyu; Mazitschek, Ralph; Bradner, James E.; DePinho, Ronald A.; Jaenisch, Rudolf; Tsai, Li-Huei

    2012-01-01

    Chromatin modifications, especially histone-tail acetylation, have been implicated in memory formation. Increased histone-tail acetylation induced by inhibitors of histone deacetylases (HDACis) facilitates learning and memory in wildtype mice as well as in mouse models of neurodegeneration. Harnessing the therapeutic potential of HDACi requires knowledge of the specific HDAC family member(s) linked to cognitive enhancement. Here we show that neuron-specific overexpression of HDAC2, but not HDAC1, reduced dendritic spine density, synapse number, synaptic plasticity, and memory formation. Conversely, HDAC2 deficiency resulted in increased synapse number and memory facilitation, similar to chronic HDACi treatment in mice. Notably, reduced synapse number and learning impairment of HDAC2-overexpressing mice were ameliorated by chronic HDACi treatment. Correspondingly, HDACi treatment failed to further facilitate memory formation in HDAC2-deficient mice. Furthermore, analysis of promoter occupancy revealed association of HDAC2 with the promoters of genes implicated in synaptic plasticity and memory formation. Together, our results suggest that HDAC2 plays a role in modulating synaptic plasticity and long-lasting changes of neural circuits, which in turn negatively regulates learning and memory. These observations encourage the development and testing of HDAC2-selective inhibitors for human diseases associated with memory impairment. PMID:19424149

  15. Actin filament curvature biases branching direction

    NASA Astrophysics Data System (ADS)

    Wang, Evan; Risca, Viviana; Chaudhuri, Ovijit; Chia, Jia-Jun; Geissler, Phillip; Fletcher, Daniel

    2012-02-01

    Actin filaments are key components of the cellular machinery, vital for a wide range of processes ranging from cell motility to endocytosis. Actin filaments can branch, and essential in this process is a protein complex known as the Arp2/3 complex, which nucleate new ``daughter'' filaments from pre-existing ``mother'' filaments by attaching itself to the mother filament. Though much progress has been made in understanding the Arp2/3-actin junction, some very interesting questions remain. In particular, F-actin is a dynamic polymer that undergoes a wide range of fluctuations. Prior studies of the Arp2/3-actin junction provides a very static notion of Arp2/3 binding. The question we ask is how differently does the Arp2/3 complex interact with a straight filament compared to a bent filament? In this study, we used Monte Carlo simulations of a surface-tethered worm-like chain to explore possible mechanisms underlying the experimental observation that there exists preferential branch formation by the Arp2/3 complex on the convex face of a curved filament. We show that a fluctuation gating model in which Arp2/3 binding to the actin filament is dependent upon a rare high-local-curvature shape fluctuation of the filament is consistent with the experimental data.

  16. Actin filament curvature biases branching direction

    PubMed Central

    Risca, Viviana I.; Wang, Evan B.; Chaudhuri, Ovijit; Chia, Jia Jun; Geissler, Phillip L.; Fletcher, Daniel A.

    2012-01-01

    Mechanical cues affect many important biological processes in metazoan cells, such as migration, proliferation, and differentiation. Such cues are thought to be detected by specialized mechanosensing molecules linked to the cytoskeleton, an intracellular network of protein filaments that provide mechanical rigidity to the cell and drive cellular shape change. The most abundant such filament, actin, forms branched networks nucleated by the actin-related protein (Arp) 2/3 complex that support or induce membrane protrusions and display adaptive behavior in response to compressive forces. Here we show that filamentous actin serves in a mechanosensitive capacity itself, by biasing the location of actin branch nucleation in response to filament bending. Using an in vitro assay to measure branching from curved sections of immobilized actin filaments, we observed preferential branch formation by the Arp2/3 complex on the convex face of the curved filament. To explain this behavior, we propose a fluctuation gating model in which filament binding or branch nucleation by Arp2/3 occur only when a sufficiently large, transient, local curvature fluctuation causes a favorable conformational change in the filament, and we show with Monte Carlo simulations that this model can quantitatively account for our experimental data. We also show how the branching bias can reinforce actin networks in response to compressive forces. These results demonstrate how filament curvature can alter the interaction of cytoskeletal filaments with regulatory proteins, suggesting that direct mechanotransduction by actin may serve as a general mechanism for organizing the cytoskeleton in response to force. PMID:22308368

  17. Femtosecond Laser Filamentation for Atmospheric Sensing

    PubMed Central

    Xu, Huai Liang; Chin, See Leang

    2011-01-01

    Powerful femtosecond laser pulses propagating in transparent materials result in the formation of self-guided structures called filaments. Such filamentation in air can be controlled to occur at a distance as far as a few kilometers, making it ideally suited for remote sensing of pollutants in the atmosphere. On the one hand, the high intensity inside the filaments can induce the fragmentation of all matters in the path of filaments, resulting in the emission of characteristic fluorescence spectra (fingerprints) from the excited fragments, which can be used for the identification of various substances including chemical and biological species. On the other hand, along with the femtosecond laser filamentation, white-light supercontinuum emission in the infrared to UV range is generated, which can be used as an ideal light source for absorption Lidar. In this paper, we present an overview of recent progress concerning remote sensing of the atmosphere using femtosecond laser filamentation. PMID:22346566

  18. Zyxin, a Regulator of Actin Filament Assembly, Targets the Mitotic Apparatus by Interacting with H-Warts/Lats1 Tumor Suppressor

    PubMed Central

    Hirota, Toru; Morisaki, Tetsuro; Nishiyama, Yasuyuki; Marumoto, Tomotoshi; Tada, Kenji; Hara, Toshihiro; Masuko, Norio; Inagaki, Masaki; Hatakeyama, Katsuyoshi; Saya, Hideyuki

    2000-01-01

    The mitotic apparatus plays a pivotal role in dividing cells to ensure each daughter cell receives a full set of chromosomes and complement of cytoplasm during mitosis. A human homologue of the Drosophila warts tumor suppressor, h-warts/LATS1, is an evolutionarily conserved serine/threonine kinase and a dynamic component of the mitotic apparatus. We have identified an interaction of h-warts/LATS1 with zyxin, a regulator of actin filament assembly. Zyxin is a component of focal adhesion, however, during mitosis a fraction of cytoplasmic-dispersed zyxin becomes associated with h-warts/LATS1 on the mitotic apparatus. We found that zyxin is phosphorylated specifically during mitosis, most likely by Cdc2 kinase, and that the phosphorylation regulates association with h-warts/LATS1. Furthermore, microinjection of truncated h-warts/LATS1 protein, including the zyxin-binding portion, interfered with localization of zyxin to mitotic apparatus, and the duration of mitosis of these injected cells was significantly longer than that of control cells. These findings suggest that h-warts/LATS1 and zyxin play a crucial role in controlling mitosis progression by forming a regulatory complex on mitotic apparatus. PMID:10831611

  19. Rictor/mTORC2 regulates blood-testis barrier dynamics via its effects on gap junction communications and actin filament network.

    PubMed

    Mok, Ka-Wai; Mruk, Dolores D; Lee, Will M; Cheng, C Yan

    2013-03-01

    In the mammalian testis, coexisting tight junctions (TJs), basal ectoplasmic specializations, and gap junctions (GJs), together with desmosomes near the basement membrane, constitute the blood-testis barrier (BTB). The most notable feature of the BTB, however, is the extensive network of actin filament bundles, which makes it one of the tightest blood-tissue barriers. The BTB undergoes restructuring to facilitate the transit of preleptotene spermatocytes at stage VIII-IX of the epithelial cycle. Thus, the F-actin network at the BTB undergoes cyclic reorganization via a yet-to-be explored mechanism. Rictor, the key component of mTORC2 that is known to regulate actin cytoskeleton, was shown to express stage-specifically at the BTB in the seminiferous epithelium. Its expression was down-regulated at the BTB in stage VIII-IX tubules, coinciding with BTB restructuring at these stages. Using an in vivo model, a down-regulation of rictor at the BTB was also detected during adjudin-induced BTB disruption, illustrating rictor expression is positively correlated with the status of the BTB integrity. Indeed, the knockdown of rictor by RNAi was found to perturb the Sertoli cell TJ-barrier function in vitro and the BTB integrity in vivo. This loss of barrier function was accompanied by changes in F-actin organization at the Sertoli cell BTB in vitro and in vivo, associated with a loss of interaction between actin and α-catenin or ZO-1. Rictor knockdown by RNAi was also found to impede Sertoli cell-cell GJ communication, disrupting protein distribution (e.g., occludin, ZO-1) at the BTB, illustrating that rictor is a crucial BTB regulator.

  20. Muscle myosin filaments: cores, crowns and couplings.

    PubMed

    Squire, John M

    2009-09-01

    Myosin filaments in muscle, carrying the ATPase myosin heads that interact with actin filaments to produce force and movement, come in multiple varieties depending on species and functional need, but most are based on a common structural theme. The now successful journeys to solve the ultrastructures of many of these myosin filaments, at least at modest resolution, have not been without their false starts and erroneous sidetracks, but the picture now emerging is of both diversity in the rotational symmetries of different filaments and a degree of commonality in the way the myosin heads are organised in resting muscle. Some of the remaining differences may be associated with how the muscle is regulated. Several proteins in cardiac muscle myosin filaments can carry mutations associated with heart disease, so the elucidation of myosin filament structure to understand the effects of these mutations has a clear and topical clinical relevance.

  1. Regulation of Memory Formation by the Transcription Factor XBP1.

    PubMed

    Martínez, Gabriela; Vidal, René L; Mardones, Pablo; Serrano, Felipe G; Ardiles, Alvaro O; Wirth, Craig; Valdés, Pamela; Thielen, Peter; Schneider, Bernard L; Kerr, Bredford; Valdés, Jose L; Palacios, Adrian G; Inestrosa, Nibaldo C; Glimcher, Laurie H; Hetz, Claudio

    2016-02-16

    Contextual memory formation relies on the induction of new genes in the hippocampus. A polymorphism in the promoter of the transcription factor XBP1 was identified as a risk factor for Alzheimer's disease and bipolar disorders. XBP1 is a major regulator of the unfolded protein response (UPR), mediating adaptation to endoplasmic reticulum (ER) stress. Using a phenotypic screen, we uncovered an unexpected function of XBP1 in cognition and behavior. Mice lacking XBP1 in the nervous system showed specific impairment of contextual memory formation and long-term potentiation (LTP), whereas neuronal XBP1s overexpression improved performance in memory tasks. Gene expression analysis revealed that XBP1 regulates a group of memory-related genes, highlighting brain-derived neurotrophic factor (BDNF), a key component in memory consolidation. Overexpression of BDNF in the hippocampus reversed the XBP1-deficient phenotype. Our study revealed an unanticipated function of XBP1 in cognitive processes that is apparently unrelated to its role in ER stress. Copyright © 2016 The Authors. Published by Elsevier Inc. All rights reserved.

  2. Nitric oxide regulates neutrophil migration through microparticle formation.

    PubMed

    Nolan, Sarah; Dixon, Rachel; Norman, Keith; Hellewell, Paul; Ridger, Victoria

    2008-01-01

    The role of nitric oxide (NO) in regulating neutrophil migration has been investigated. Human neutrophil migration to interleukin (IL)-8 (1 nmol/L) was measured after a 1-hour incubation using a 96-well chemotaxis plate assay. The NO synthase inhibitor N(G)-nitro-l-arginine methyl ester (L-NAME) significantly (P < 0.001) enhanced IL-8-induced migration by up to 45%. Anti-CD18 significantly (P < 0.001) inhibited both IL-8-induced and L-NAME enhanced migration. Antibodies to L-selectin or PSGL-1 had no effect on IL-8-induced migration but prevented the increased migration to IL-8 induced by L-NAME. L-NAME induced generation of neutrophil-derived microparticles that was significantly (P < 0.01) greater than untreated neutrophils or D-NAME. This microparticle formation was dependent on calpain activity and superoxide production. Only microparticles from L-NAME and not untreated or D-NAME-treated neutrophils induced a significant (P < 0.01) increase in IL-8-induced migration and transendothelial migration. Pretreatment of microparticles with antibodies to L-selectin (DREG-200) or PSGL-1 (PL-1) significantly (P < 0.001) inhibited this effect. The ability of L-NAME-induced microparticles to enhance migration was found to be dependent on the number of microparticles produced and not an increase in microparticle surface L-selectin or PSGL-1 expression. These data show that NO can modulate neutrophil migration by regulating microparticle formation.

  3. Solubilization and fractionation of paired helical filaments.

    PubMed

    González, P J; Correas, I; Avila, J

    1992-09-01

    Paired helical filaments isolated from brains of two different patients with Alzheimer's disease were extensively treated with the ionic detergent, sodium dodecyl sulphate. Filaments were solubilized at different extents, depending on the brain examined, thus suggesting the existence of two types of paired helical filaments: sodium dodecyl sulphate-soluble and insoluble filaments. In the first case, the number of structures resembling paired helical filaments greatly decreased after the detergent treatment, as observed by electron microscopy. Simultaneously, a decrease in the amount of sedimentable protein was also observed upon centrifugation of the sodium dodecyl sulfate-treated paired helical filaments. A sodium dodecyl sulphate-soluble fraction was isolated as a supernatant after low-speed centrifugation of the sodium dodecyl sulphate-treated paired helical filaments. The addition of the non-ionic detergent Nonidet-P40 to this fraction resulted in the formation of paired helical filament-like structures. When the sodium dodecyl sulphate-soluble fraction was further fractionated by high-speed centrifugation, three subfractions were observed: a supernatant, a pellet and a thin layer between these two subfractions. No paired helical filaments were observed in any of these subfractions, even after addition of Nonidet P-40. However, when they were mixed back together, the treatment with Nonidet P-40 resulted in the visualization of paired helical filament-like structures. These results suggest that at least two different components are needed for the reconstitution of paired helical filaments as determined by electron microscopy. The method described here may allow the study of the components involved in the formation of paired helical filaments and the identification of possible factors capable of blocking this process.

  4. CHIP Regulates Osteoclast Formation through Promoting TRAF6 Protein Degradation

    PubMed Central

    Li, Shan; Shu, Bing; Zhang, Yanquan; Li, Jia; Guo, Junwei; Wang, Yinyin; Ren, Fangli; Xiao, Guozhi; Chang, Zhijie; Chen, Di

    2014-01-01

    Objective Carboxyl terminus of Hsp70-interacting protein (CHIP or STUB1) is an E3 ligase and regulates the stability of several proteins which are involved in tumor growth and metastasis. However, the role of CHIP in bone growth and bone remodeling in vivo has not been reported. The objective of this study is to investigate the role and mechanism of CHIP in regulation of bone mass and bone remodeling. Methods The bone phenotype of Chip−/− mice was examined by histology, histomorphometry and micro-CT analyses. The regulatory mechanism of CHIP on the degradation of TRAF6 and the inhibition of NF-κB signaling was examined by immunoprecipitation (IP), western blotting and luciferase reporter assays. Results In this study, we found that deletion of the Chip gene leads to osteopenic phenotype and increased osteoclast formation. We further found that TRAF6, as a novel substrate of CHIP, is up-regulated in Chip−/− osteoclasts. TRAF6 is critical for RANKL-induced osteoclastogenesis. TRAF6 is an adaptor protein which functions as an E3 ligase to regulate the activation of TAK1 and the I-κB kinase (IKK) and is a key regulator of NF-κB signaling. CHIP interacts with TRAF6 to promote TRAF6 ubiquitination and proteasome degradation. CHIP inhibits p65 nuclear translocation, leading to the repression of the TRAF6-mediated NF-κB transcription. Conclusion CHIP inhibits NF-κB signaling via promoting TRAF6 degradation and plays an important role in osteoclastogenesis and bone remodeling, suggesting that it may be a novel therapeutic target for the treatment of bone loss associated diseases. PMID:24578159

  5. The effects of transverse plasma flow on laser beam deflection and of ultra-intense laser beam filamentation on channel formation

    NASA Astrophysics Data System (ADS)

    Hinkel, D. E.

    1997-11-01

    Recent experiments conducted at Lawrence Livermore National Laboratory (LLNL) with the Nova and Janus lasers demonstrate deflection of the laser beam in plasma with flow transverse to the beam. In gas-filled hohlraum experiments(S. G. Glendinning et al.), the laser spot on the hohlraum wall is ~ 100 μm closer to the laser entrance hole (LEH) than in empty hohlraum experiments, which degrades drive symmetry. In a series of exploding foil experiments(J. D. Moody et al.), Phys. Rev. Lett. 77, 1294 (1996)., intensity dependent deflection of the transmitted beam is observed, and interferometric measurements of laser-produced channels in preformed plasma(P. E. Young et al.), to be submitted to Phys. Rev. Lett., 1997. show beam deflection in the presence of near-sonic transverse flow. Theoretical analysis(D. E. Hinkel et al.), Phys. Rev. Lett. 77, 1298 (1996). yields simple scaling laws for the formation of ponderomotively (or thermally) created density depressions downstream from the laser beam's high intensity regions, into which the light is refracted. An integrated approach that utilizes plasma parameters from the hydrocode Lasnex, detailed knowledge of the beam structure, and plasma physics analysis and modelling with F3D(R. L. Berger et al.), Phys. Fluids B 5, 2243 (1993)., has been used to develop a predictive capability that successfully quantifies the amount of beam deflection occurring in experiments. Related physics of beam self-focussing and filamentation is of relevance to the Fast Ignitor(M. Tabak et al.), Phys. Plasmas 1, 1626 (1994).. In channeling experiments performed on the 100 TW laser at LLNL, the f/3 laser beam, which has a 15 μm waist at best focus, has intensities in excess of IL = 1 × 10^17 W/cm^2. Modelling of these high intensity experiments indicates that channel formation occurs over a wide range of cone angles for an idealized (Gaussian) beam. However, when beam structure is taken into consideration, channel formation in the underdense

  6. Supramolecular Assembly in Cytoskeletal Filaments and their Associated Biomolecules

    NASA Astrophysics Data System (ADS)

    Safinya, Cyrus R.

    2002-03-01

    With the completion of the Human Genome Project and the emerging proteomics era, the biosciences community is beginning the daunting task of understanding the functions of a large number of interacting proteins. Cellular activity, which is usually tightly regulated, results from protein-protein and protein-nucleic acid interactions, which often lead to the formation of very large assemblies of biomolecules for distinct functions. Examples include DNA condensation states during the cell cycle, and bundle and network formation of filamentous proteins in cell attachment, motility, and cytokinesis. We present recent synchrotron x-ray diffraction and optical imaging data, in cell-free systems of cytoskeletal filaments and their associated biomolecules, which reveal novel supramolecular assemblies, spanning lengths from the nanometer to the micrometer scale. Supported by NSF DMR-9972246 and NIH GM59288.

  7. Engineering transcriptional regulation to control Pdu microcompartment formation.

    PubMed

    Kim, Edward Y; Jakobson, Christopher M; Tullman-Ercek, Danielle

    2014-01-01

    Bacterial microcompartments (MCPs) show great promise for the organization of engineered metabolic pathways within the bacterial cytoplasm. This subcellular organelle is composed of a protein shell of 100-200 nm diameter that natively encapsulates multi-enzyme pathways. The high energy cost of synthesizing the thousands of protein subunits required for each MCP demands precise regulation of MCP formation for both native and engineered systems. Here, we study the regulation of the propanediol utilization (Pdu) MCP, for which growth on 1,2-propanediol induces expression of the Pdu operon for the catabolism of 1,2-propanediol. We construct a fluorescence-based transcriptional reporter to investigate the activation of the Ppdu promoter, which drives the transcription of 21 pdu genes. Guided by this reporter, we find that MCPs can be expressed in strains grown in rich media, provided that glucose is not present. We also characterize the response of the Ppdu promoter to a transcriptional activator of the pdu operon, PocR, and find PocR to be a necessary component of Pdu MCP formation. Furthermore, we find that MCPs form normally upon the heterologous expression of PocR even in the absence of the natural inducer 1,2-propanediol and in the presence of glucose, and that Pdu MCPs formed in response to heterologous PocR expression can metabolize 1,2-propanediol in vivo. We anticipate that this technique of overexpressing a key transcription factor may be used to study and engineer the formation, size, and/or number of MCPs for the Pdu and related MCP systems.

  8. Engineering Transcriptional Regulation to Control Pdu Microcompartment Formation

    PubMed Central

    Tullman-Ercek, Danielle

    2014-01-01

    Bacterial microcompartments (MCPs) show great promise for the organization of engineered metabolic pathways within the bacterial cytoplasm. This subcellular organelle is composed of a protein shell of 100–200 nm diameter that natively encapsulates multi-enzyme pathways. The high energy cost of synthesizing the thousands of protein subunits required for each MCP demands precise regulation of MCP formation for both native and engineered systems. Here, we study the regulation of the propanediol utilization (Pdu) MCP, for which growth on 1,2-propanediol induces expression of the Pdu operon for the catabolism of 1,2-propanediol. We construct a fluorescence-based transcriptional reporter to investigate the activation of the Ppdu promoter, which drives the transcription of 21 pdu genes. Guided by this reporter, we find that MCPs can be expressed in strains grown in rich media, provided that glucose is not present. We also characterize the response of the Ppdu promoter to a transcriptional activator of the pdu operon, PocR, and find PocR to be a necessary component of Pdu MCP formation. Furthermore, we find that MCPs form normally upon the heterologous expression of PocR even in the absence of the natural inducer 1,2-propanediol and in the presence of glucose, and that Pdu MCPs formed in response to heterologous PocR expression can metabolize 1,2-propanediol in vivo. We anticipate that this technique of overexpressing a key transcription factor may be used to study and engineer the formation, size, and/or number of MCPs for the Pdu and related MCP systems. PMID:25427074

  9. Transcriptional regulation of mouse hypoglossal motor neuron somatotopic map formation.

    PubMed

    Chen, Xin; Wang, Jae Woong; Salin-Cantegrel, Adele; Dali, Rola; Stifani, Stefano

    2016-11-01

    Somatic motor neurons in the hypoglossal nucleus innervate tongue muscles controlling vital functions such as chewing, swallowing and respiration. Formation of functional hypoglossal nerve circuits depends on the establishment of precise hypoglossal motor neuron maps correlating with specific tongue muscle innervations. Little is known about the molecular mechanisms controlling mammalian hypoglossal motor neuron topographic map formation. Here we show that combinatorial expression of transcription factors Runx1, SCIP and FoxP1 defines separate mouse hypoglossal motor neuron groups with different topological, neurotransmitter and calcium-buffering phenotypes. Runx1 and SCIP are coexpressed in ventromedial hypoglossal motor neurons involved in control of tongue protrusion whereas FoxP1 is expressed in dorsomedial motor neurons associated with tongue retraction. Establishment of separate hypoglossal motor neuron maps depends in part on Runx1-mediated suppression of ventrolateral and dorsomedial motor neuron phenotypes and regulation of FoxP1 expression pattern. These findings suggest that combinatorial actions of Runx1, SCIP and FoxP1 are important for mouse hypoglossal nucleus somatotopic map formation.

  10. Structural characterization of human vimentin rod 1 and the sequencing of assembly steps in intermediate filament formation in vitro using site-directed spin labeling and electron paramagnetic resonance.

    PubMed

    Hess, John F; Budamagunta, Madhu S; Voss, John C; FitzGerald, Paul G

    2004-10-22

    We have previously established the utility of site-directed spin labeling and electron paramagnetic resonance to determine structural relationships among proteins in intact intermediate filaments. Using this same approach we have introduced spin labels at 21 residues between amino acids 169 and 193 in rod domain 1 of human vimentin. The electron paramagnetic resonance spectra provide direct evidence for the coiled coil nature of the vimentin dimer in this region. This finding is consistent with predictions but has never been demonstrated previously. In a previous study we identified residue 348 in the rod domain 2 as one point of overlap between adjacent dimers in intact filaments. In the present study we defined residue 191 in the rod domain 1 as a second point of overlap and established that the dimers are arranged in an anti-parallel and staggered orientation at this site. Finally, by isolating spin-labeled samples at successive stages during the dialysis that lead to filament assembly in vitro, we have been able to establish a sequence of interactions that occurs during in vitro assembly, starting with the alpha helix and loose coiled coil dimer formation, then the formation of tetrameric species centered on residue 191, followed by interactions centered on residue 348 suggestive of octamer or higher order multimer formation. A continuation of this strategy revealed that both 191-191 and 348-348 interactions are present in low ionic strength Tris buffers when vimentin is maintained at the "protofilament" stage of assembly.

  11. Regulation of Bone Formation During Disuse by miRNA

    NASA Technical Reports Server (NTRS)

    Thomas, Nicholas; Choi, Catherine Y.; Alwood, Joshua S.

    2016-01-01

    Astronauts lose bone structure during long-duration spaceflight. These changes are due, in part, to insufficient bone formation by the osteoblast cells. Little is known about the role that small (approximately 22 nucleotide), non-coding micro-RNAs (miRNAs) play in the osteoblast response to microgravity. We hypothesize that osteoblast-lineage cells alter their miRNA status during microgravity exposure, contributing to impaired bone formation during weightlessness. To simulate weightlessness, female mice (C57BL/6, Charles River, 10 weeks of age, n = 6) were hindlimb unloaded for 12 days. Age-matched and normally ambulating mice served as controls (n=6). To assess the expression of miRNAs in skeletal tissue, the right and left tibia of the mice were collected ex vivo and cleaned of soft-tissue and marrow. Total RNA was collected from tibial bone and relative abundance was measured for miRNAs of interest using quantitative real time PCR array looking at 372 unique and well-characterized mature miRNAs using the delta-delta Ct method. Transcripts of interest were normalized to an average of 6 reference RNAs. Preliminary results show that hindlimb unloading decreased the expression of 14 miRNAs to less than 1.4-2.9X control levels and increased the expression of 5 miRNAs relative to the control mice greater than 1-2-1.5X (p less than 0.05, respectively). Using the miRSystem we assessed overlapping target genes predicted to be regulated by multiple members of the 19 differentially expressed miRNAs as well as in silico predicted targets of our individual miRNAs. Our miRSystem results indicated that a number of our differentially expressed miRNAs were regulators of genes related to the Wnt-Beta Catenin pathway-a known regulator of bone health-and, interestingly, the estrogen-mediated cell-cycle regulation pathway, which may indicate that simulated weightlessness induced systemic hormonal changes that contributed to bone loss. We plan to follow up these findings by measuring

  12. Regulation of contractile vacuole formation and activity in Dictyostelium

    PubMed Central

    Du, Fei; Edwards, Kimberly; Shen, Zhouxin; Sun, Binggang; De Lozanne, Arturo; Briggs, Steven; Firtel, Richard A

    2008-01-01

    The contractile vacuole (CV) system is the osmoregulatory organelle required for survival for many free-living cells under hypotonic conditions. We identified a new CV regulator, Disgorgin, a TBC-domain-containing protein, which translocates to the CV membrane at the late stage of CV charging and regulates CV–plasma membrane fusion and discharging. disgorgin− cells produce large CVs due to impaired CV–plasma membrane fusion. Disgorgin is a specific GAP for Rab8A-GTP, which also localizes to the CV and whose hydrolysis is required for discharging. We demonstrate that Drainin, a previously identified TBC-domain-containing protein, lies upstream from Disgorgin in this pathway. Unlike Disgorgin, Drainin lacks GAP activity but functions as a Rab11A effector. The BEACH family proteins LvsA and LvsD were identified in a suppressor/enhancer screen of the disgorgin− large CV phenotype and demonstrated to have distinct functions in regulating CV formation. Our studies help define the pathways controlling CV function. PMID:18636095

  13. Repo-Man/PP1 regulates heterochromatin formation in interphase

    PubMed Central

    de Castro, Inês J.; Budzak, James; Di Giacinto, Maria L.; Ligammari, Lorena; Gokhan, Ezgi; Spanos, Christos; Moralli, Daniela; Richardson, Christine; de las Heras, Jose I.; Salatino, Silvia; Schirmer, Eric C.; Ullman, Katharine S.; Bickmore, Wendy A.; Green, Catherine; Rappsilber, Juri; Lamble, Sarah; Goldberg, Martin W.; Vinciotti, Veronica; Vagnarelli, Paola

    2017-01-01

    Repo-Man is a protein phosphatase 1 (PP1) targeting subunit that regulates mitotic progression and chromatin remodelling. After mitosis, Repo-Man/PP1 remains associated with chromatin but its function in interphase is not known. Here we show that Repo-Man, via Nup153, is enriched on condensed chromatin at the nuclear periphery and at the edge of the nucleopore basket. Repo-Man/PP1 regulates the formation of heterochromatin, dephosphorylates H3S28 and it is necessary and sufficient for heterochromatin protein 1 binding and H3K27me3 recruitment. Using a novel proteogenomic approach, we show that Repo-Man is enriched at subtelomeric regions together with H2AZ and H3.3 and that depletion of Repo-Man alters the peripheral localization of a subset of these regions and alleviates repression of some polycomb telomeric genes. This study shows a role for a mitotic phosphatase in the regulation of the epigenetic landscape and gene expression in interphase. PMID:28091603

  14. Cyclic Expression of Lhx2 Regulates Hair Formation

    PubMed Central

    Hägglund, Anna-Carin; Carlsson, Leif

    2010-01-01

    Hair is important for thermoregulation, physical protection, sensory activity, seasonal camouflage, and social interactions. Hair is generated in hair follicles (HFs) and, following morphogenesis, HFs undergo cyclic phases of active growth (anagen), regression (catagen), and inactivity (telogen) throughout life. The transcriptional regulation of this process is not well understood. We show that the transcription factor Lhx2 is expressed in cells of the outer root sheath and a subpopulation of matrix cells during both morphogenesis and anagen. As the HFs enter telogen, expression becomes undetectable and reappears prior to initiation of anagen in the secondary hair germ. In contrast to previously published results, we find that Lhx2 is primarily expressed by precursor cells outside of the bulge region where the HF stem cells are located. This developmental, stage- and cell-specific expression suggests that Lhx2 regulates the generation and regeneration of hair. In support of this hypothesis, we show that Lhx2 is required for anagen progression and HF morphogenesis. Moreover, transgenic expression of Lhx2 in postnatal HFs is sufficient to induce anagen. Thus, our results reveal an alternative interpretation of Lhx2 function in HFs compared to previously published results, since Lhx2 is periodically expressed, primarily in precursor cells distinct from those in the bulge region, and is an essential positive regulator of hair formation. PMID:20386748

  15. Environment and self-regulation in galaxy formation

    NASA Astrophysics Data System (ADS)

    Thomas, Daniel; Maraston, Claudia; Schawinski, Kevin; Sarzi, Marc; Silk, Joseph

    2010-06-01

    The environment is known to affect the formation and evolution of galaxies considerably best visible through the well-known morphology-density relationship. It is less clear, though, whether the environment is equally important at a given galaxy morphology. In this paper, we study the effect of environment on the evolution of early-type galaxies as imprinted in the fossil record by analysing the stellar population properties of 3360 galaxies morphologically selected by visual inspection from the Sloan Digital Sky Survey in a narrow redshift range (0.05 <= z <= 0.06). The morphological selection algorithm is critical, as it does not bias against recent star formation. We find that the distribution of ages is bimodal with a strong peak at old ages and a secondary peak at young ages around ~2.5Gyr containing about 10 per cent of the objects. This is an analogue to `red sequence' and `blue cloud' identified in galaxy populations usually containing both early- and late-type galaxies. The fraction of the young, rejuvenated galaxies increases with both decreasing galaxy mass and decreasing environmental density up to about 45 per cent, which implies that the impact of environment increases with decreasing galaxy mass. The rejuvenated galaxies have lower α/Fe ratios than the average and most of them show signs of ongoing star formation through their emission line spectra. All objects that host active galactic nuclei in their centres without star formation are part of the red sequence population. We confirm and statistically strengthen earlier results that luminosity weighted ages, metallicities and α/Fe element ratios of the red sequence population correlate well with velocity dispersion and galaxy mass. Most interestingly, however, these scaling relations are not sensitive to environmental densities and are only driven by galaxy mass. We infer that early-type galaxy formation has undergone a phase transition a few billion years ago around z ~ 0.2. A self-regulated

  16. Effect of Actin Filament on Deformation-Induced Ca2+ Response in Osteoblast-Like Cells

    NASA Astrophysics Data System (ADS)

    Adachi, Taiji; Murai, Takayuki; Hoshiai, Sodai; Tomita, Yoshihiro

    Under the influence of mechanical environment, bone structure is formed and maintained by adaptive remodeling that involves osteoclastic resorption and osteoblastic formation. In the mechanotransduction system in osteoblasts, it is believed that intracellular calcium plays a fundamental role and cytoskeletal actin filament is a crucial component for the signal transduction process. To clarify the role of actin filament in deformation-induced Ca2+ signaling, osteoblast-like cells (MC3T3-E1) with different actin filament densities controlled by cytochalasin D were subjected to tensile strain in vitro. The change in intracellular Ca2+ concentration labeled by fluo-3 was observed using a confocal laser-scanning microscope. As a result, the disruption of the actin filament was found to significantly suppress the deformation-induced Ca2+ response that was regulated according to the degree of actin filament organization. This result indicates that the actin filament is indispensable for the quantitative regulation of Ca2+ signaling in response to a mechanical stimulus in osteoblasts.

  17. Characterization and regulation of an additional actin-filament-binding site in large isoforms of the stereocilia actin-bundling protein espin.

    PubMed

    Zheng, Lili; Beeler, Dina M; Bartles, James R

    2014-03-15

    The espin actin-bundling proteins, which are produced as isoforms of different sizes from a single gene, are required for the growth of hair cell stereocilia. We have characterized an additional actin-filament-binding site present in the extended amino-termini of large espin isoforms. Constitutively active in espin 2, the site increased the size of actin bundles formed in vitro and inhibited actin fluorescence recovery in microvilli. In espin 1, which has an N-terminal ankyrin repeat domain, the site was autoinhibited by binding between the ankyrin repeat domain and a peptide near the actin-binding site. Deletion of this peptide from espin 1 activated its actin-binding site. The peptide resembled tail homology domain I of myosin III, a ligand of the ankyrin repeat domain localized with espin 1 at the tip of stereocilia. A myosin III tail homology domain I peptide, but not scrambled control peptides, inhibited internal binding of the ankyrin repeat domain and released the espin 1 actin-binding site from autoinhibition. Thus, this regulation could result in local activation of the additional actin-binding site of espin 1 by myosin III in stereocilia.

  18. Regulation of platelet plug formation by phosphoinositide metabolism

    PubMed Central

    Min, Sang H.

    2013-01-01

    Phosphatidylinositol and its phosphorylated derivatives, phosphoinositides, are minor constituents of phospholipids at the cellular membrane level. Nevertheless, phosphatidylinositol and phosphoinositides represent essential components of intracellular signaling that regulate diverse cellular processes, including platelet plug formation. Accumulating evidence indicates that the metabolism of phosphoinositides is temporally and spatially modulated by the opposing effects of specific phosphoinositide-metabolizing enzymes, including lipid kinases, lipid phosphatases, and phospholipases. Each of these enzymes generates a selective phosphoinositide or second messenger within precise cellular compartments. Intriguingly, phosphoinositide-metabolizing enzymes exist in different isoforms, which all produce the same phosphoinositide products. Recent studies using isoform-specific mouse models and chemical inhibitors have elucidated that the different isoforms of phosphoinositide-metabolizing enzymes have nonredundant functions and provide an additional layer of complexity to the temporo-spatial organization of intracellular signaling events. In this review, we will discuss recent advances in our understanding of phosphoinositide organization during platelet activation. PMID:23757731

  19. Survivin isoform Delta Ex3 regulates tumor spheroid formation.

    PubMed

    Espinosa, Magali; Ceballos-Cancino, Gisela; Callaghan, Richard; Maldonado, Vilma; Patiño, Nelly; Ruíz, Víctor; Meléndez-Zajgla, Jorge

    2012-05-01

    Survivin is an important member of the Inhibitor of Apoptosis Proteins (IAPs) family and has essential roles in apoptosis and cell cycle progression. This gene is commonly upregulated in human cancer and provides an exciting diagnostic and therapeutic target. Survivin is expressed as several isoforms that are generated by alternative splicing, and some of these present antagonistic activities. Currently, information regarding the regulation of these isoforms is lacking. In this study, we sought to analyze survivin Delta Ex3 expression in a three-dimensional model of avascular tumors and its overexpression effects in processes such as proliferation, clonogenicity and apoptosis. We found a positive correlation between spheroid growth and survivin Delta Ex3 expression during the exponential phase. We demonstrated that this isoform not only decreased apoptosis but also inhibited tumor spheroid formation by decreasing proliferation and clonogenic survival. These results point toward a dual and antagonistic effect of this spliced survivin isoform in cancer development.

  20. Role of Active Contraction and Tropomodulins in Regulating Actin Filament Length and Sarcomere Structure in Developing Zebrafish Skeletal Muscle

    PubMed Central

    Mazelet, Lise; Parker, Matthew O.; Li, Mei; Arner, Anders; Ashworth, Rachel

    2016-01-01

    Whilst it is recognized that contraction plays an important part in maintaining the structure and function of mature skeletal muscle, its role during development remains undefined. In this study the role of movement in skeletal muscle maturation was investigated in intact zebrafish embryos using a combination of genetic and pharmacological approaches. An immotile mutant line (cacnb1ts25) which lacks functional voltage-gated calcium channels (dihydropyridine receptors) in the muscle and pharmacological immobilization of embryos with a reversible anesthetic (Tricaine), allowed the study of paralysis (in mutants and anesthetized fish) and recovery of movement (reversal of anesthetic treatment). The effect of paralysis in early embryos (aged between 17 and 24 hours post-fertilization, hpf) on skeletal muscle structure at both myofibrillar and myofilament level was determined using both immunostaining with confocal microscopy and small angle X-ray diffraction. The consequences of paralysis and subsequent recovery on the localization of the actin capping proteins Tropomodulin 1 & 4 (Tmod) in fish aged from 17 hpf until 42 hpf was also assessed. The functional consequences of early paralysis were investigated by examining the mechanical properties of the larval muscle. The length-force relationship, active and passive tension, was measured in immotile, recovered and control skeletal muscle at 5 and 7 day post-fertilization (dpf). Recovery of muscle function was also assessed by examining swimming patterns in recovered and control fish. Inhibition of the initial embryonic movements (up to 24 hpf) resulted in an increase in myofibril length and a decrease in width followed by almost complete recovery in both moving and paralyzed fish by 42 hpf. In conclusion, myofibril organization is regulated by a dual mechanism involving movement-dependent and movement-independent processes. The initial contractile event itself drives the localization of Tmod1 to its sarcomeric position

  1. Thyrostimulin Regulates Osteoblastic Bone Formation During Early Skeletal Development

    PubMed Central

    van der Spek, Anne; Logan, John G.; Gogakos, Apostolos; Bagchi-Chakraborty, Jayashree; Murphy, Elaine; van Zeijl, Clementine; Down, Jenny; Croucher, Peter I.; Boyde, Alan; Boelen, Anita

    2015-01-01

    The ancestral glycoprotein hormone thyrostimulin is a heterodimer of unique glycoprotein hormone subunit alpha (GPA)2 and glycoprotein hormone subunit beta (GPB)5 subunits with high affinity for the TSH receptor. Transgenic overexpression of GPB5 in mice results in cranial abnormalities, but the role of thyrostimulin in bone remains unknown. We hypothesized that thyrostimulin exerts paracrine actions in bone and determined: 1) GPA2 and GPB5 expression in osteoblasts and osteoclasts, 2) the skeletal consequences of thyrostimulin deficiency in GPB5 knockout (KO) mice, and 3) osteoblast and osteoclast responses to thyrostimulin treatment. Gpa2 and Gpb5 expression was identified in the newborn skeleton but declined rapidly thereafter. GPA2 and GPB5 mRNAs were also expressed in primary osteoblasts and osteoclasts at varying concentrations. Juvenile thyrostimulin-deficient mice had increased bone volume and mineralization as a result of increased osteoblastic bone formation. However, thyrostimulin failed to induce a canonical cAMP response or activate the noncanonical Akt, ERK, or mitogen-activated protein kinase (P38) signaling pathways in primary calvarial or bone marrow stromal cell-derived osteoblasts. Furthermore, thyrostimulin did not directly inhibit osteoblast proliferation, differentiation or mineralization in vitro. These studies identify thyrostimulin as a negative but indirect regulator of osteoblastic bone formation during skeletal development. PMID:26018249

  2. Cellular Regulation of Amyloid Formation in Aging and Disease

    PubMed Central

    Stroo, Esther; Koopman, Mandy; Nollen, Ellen A. A.; Mata-Cabana, Alejandro

    2017-01-01

    As the population is aging, the incidence of age-related neurodegenerative diseases, such as Alzheimer and Parkinson disease, is growing. The pathology of neurodegenerative diseases is characterized by the presence of protein aggregates of disease specific proteins in the brain of patients. Under certain conditions these disease proteins can undergo structural rearrangements resulting in misfolded proteins that can lead to the formation of aggregates with a fibrillar amyloid-like structure. Cells have different mechanisms to deal with this protein aggregation, where the molecular chaperone machinery constitutes the first line of defense against misfolded proteins. Proteins that cannot be refolded are subjected to degradation and compartmentalization processes. Amyloid formation has traditionally been described as responsible for the proteotoxicity associated with different neurodegenerative disorders. Several mechanisms have been suggested to explain such toxicity, including the sequestration of key proteins and the overload of the protein quality control system. Here, we review different aspects of the involvement of amyloid-forming proteins in disease, mechanisms of toxicity, structural features, and biological functions of amyloids, as well as the cellular mechanisms that modulate and regulate protein aggregation, including the presence of enhancers and suppressors of aggregation, and how aging impacts the functioning of these mechanisms, with special attention to the molecular chaperones. PMID:28261044

  3. Regulation of scar formation by vascular endothelial growth factor

    PubMed Central

    Wilgus, Traci A.; Ferreira, Ahalia M.; Oberyszyn, Tatiana M.; Bergdall, Valerie K.; DiPietro, Luisa A.

    2009-01-01

    Vascular endothelial growth factor (VEGF-A) is known for its effects on endothelial cells and as a positive mediator of angiogenesis. VEGF is thought to promote the repair of cutaneous wounds due to its pro-angiogenic properties, but its ability to regulate other aspects of wound repair, such as the generation of scar tissue has not been well studied. We examined the role of VEGF in scar tissue production utilizing models of scarless and fibrotic repair. Scarless fetal wounds had lower levels of VEGF and were less vascular than fibrotic fetal wounds, and the scarless phenotype could be converted to a scar-forming phenotype by adding exogenous VEGF. Similarly, neutralization of VEGF reduced vascularity and decreased scar formation in adult wounds. These results show that VEGF levels have a strong influence on scar tissue formation. Our data suggest that VEGF may not simply function as a mediator of wound angiogenesis, but instead may play a more diverse role in the wound repair process. PMID:18427552

  4. Estrogen regulates pulmonary alveolar formation, loss, and regeneration in mice.

    PubMed

    Massaro, Donald; Massaro, Gloria Decarlo

    2004-12-01

    Lung tissue elastic recoil and the dimension and number of pulmonary gas-exchange units (alveoli) are major determinants of gas-exchange function. Loss of gas-exchange function accelerates after menopause in the healthy aged and is progressively lost in individuals with chronic obstructive pulmonary disease (COPD). The latter, a disease of midlife and later, though more common in men than in women, is a disease to which women smokers and never smokers may be more susceptible than men; it is characterized by diminished lung tissue elastic recoil and presently irremediable alveolar loss. Ovariectomy in sexually immature rats diminishes the formation of alveoli, and estrogen prevents the diminution. In the present work, we found that estrogen receptor-alpha and estrogen receptor-beta, the only recognized mammalian estrogen receptors, are required for the formation of a full complement of alveoli in female mice. However, only the absence of estrogen receptor-beta diminishes lung elastic tissue recoil. Furthermore, ovariectomy in adult mice results, within 3 wk, in loss of alveoli and of alveolar surface area without a change of lung volume. Estrogen replacement, after alveolar loss, induces alveolar regeneration, reversing the architectural effects of ovariectomy. These studies 1) reveal estrogen receptors regulate alveolar size and number in a nonredundant manner, 2) show estrogen is required for maintenance of already formed alveoli and induces alveolar regeneration after their loss in adult ovariectomized mice, and 3) offer the possibility estrogen can slow alveolar loss and induce alveolar regeneration in women with COPD.

  5. Afferent input regulates the formation of distal dendritic branches.

    PubMed

    Mizrahi, Adi; Libersat, Frederic

    2002-10-07

    During postembryonic development, the dendritic arbors of neurons grow to accommodate new incoming synaptic inputs. Our goal was to examine which features of dendritic architecture of postsynaptic interneurons are regulated by these synaptic inputs. To address this question, we took advantage of the cockroach cercal system where the morphology of the sensory giant interneurons (GIs) is uniquely identified and, therefore, amenable to quantitative analysis. We analyzed the three-dimensional architecture of chronically deafferented vs. normally developed dendritic trees of a specific identified GI, namely GI2. GI2 shows five prominent dendrites, four of which were significantly altered after deafferentation. De-afferentation induced an average of 55% decrease in metric measures (number of branch points, total length, and total surface area) on the entire dendritic tree. Sholl and branch order analysis showed a decrease in the most distal and higher order branches. We suggest that afferent input plays a specific role in shaping the morphology of dendritic trees by regulating the formation or maintenance of high-order distal branches.

  6. Diversin regulates heart formation and gastrulation movements in development

    PubMed Central

    Moeller, Heinz; Jenny, Andreas; Schaeffer, Hans-Joerg; Schwarz-Romond, Thomas; Mlodzik, Marek; Hammerschmidt, Matthias; Birchmeier, Walter

    2006-01-01

    Canonical and noncanonical Wnt signaling regulate crucial events in the development of vertebrates and invertebrates. In this work we show that vertebrate Diversin, a potential orthologue of Drosophila Diego, controls fusion of heart precursors and gastrulation movements in zebrafish embryogenesis. These events are regulated by noncanonical Wnt signaling, which is independent of β-catenin. We found that Diversin directly interacts with Dishevelled and that this interaction is necessary and sufficient to mediate signals of the noncanonical Wnt pathway to downstream effectors like Rho family GTPases and Jun N-terminal kinase. The ankyrin repeats of Diversin are required for the interaction with Dishevelled, for the activation of noncanonical Wnt signaling, and for the biological responses. The mutation K446M in the DEP domain of vertebrate Dishevelled, which mimics a classical Drosophila loss of function mutation, prevents functional interaction with Diversin's ankyrin repeats. Diversin also affects planar cell polarity in Drosophila, which is controlled by the noncanonical Wnt signaling pathway. Our data thus demonstrate that Diversin and Dishevelled function together in a mutually dependent fashion in zebrafish gastrulation and organ formation. PMID:17032765

  7. Regulation of BMP-induced ectopic bone formation by Ahsg.

    PubMed

    Rittenberg, B; Partridge, E; Baker, G; Clokie, C; Zohar, R; Dennis, J W; Tenenbaum, H C

    2005-05-01

    alpha2-HS-glycoprotein (Ahsg), also known as fetuin is a serum and bone resident glycoprotein, which binds to TGF-beta superfamily members including bone morphogenetic proteins (BMP) and inhibits dexamethasone-induced osteogenesis in bone marrow cultures in vitro. Here we demonstrate that Ahsg reduces cytokine binding to its cognate receptor in HOS osteocyte cells and suppresses intracellular signaling, while in vivo, we test the hypothesis that Ahsg-deficient mice are hyper-responsive to BMP-induced osteogenesis. Human native BMP was implanted into the hindquarter muscles of Ahsg(+/+), Ahsg(+/-) and Ahsg(-/-) mice and 4 weeks later, ossicle formation was analyzed by radiography, bone density scanning (DEXA) and histomorphometry. Alkaline phosphatase (AP) activity was measured in ossicles as a marker for bone cell differentiation, and was significantly higher in Ahsg(-/-) versus Ahsg(+/-) and/or Ahsg(+/+) mice. Ectopic ossicle size in the Ahsg(+/-) mouse was 4-fold greater than that in the wild type (Ahsg(+/+)), and intermediate to that shown in Ahsg(-/-) mouse. Bone mineral density (BMD) was lower in the Ahsg(-/+) and Ahsg(-/-) mice compared to Ahsg(+/+) littermates. The ratio of cortical to cancellous bone was found to be >2-fold higher in Ahsg(-/-) mouse in comparison to the Ahsg(+/+) mice with no significant change in the Ahsg(-/+) mouse. Finally, a significantly higher incidence of satellite ossification; small islands of immature bone, was shown in Ahsg(-/-) mice as compared to Ahsg(+/+) mice. Although Ahsg binds to TGF-beta/BMP and blocks receptor signalling, it may also sequester cytokines in matrix, thereby acting as a reservoir of osteoinductive activity when released. This may explain the non-linear relationship between ectopic bone formation characteristics and Ahsg(+/+), Ahsg(+/-) and Ahsg(-/-) genotypes, although the increase in satellite bone formation might also explain this phenomenon. Our results suggest that Ahsg may be useful for prevention of

  8. Vitamin C: A Novel Regulator of Neutrophil Extracellular Trap Formation

    PubMed Central

    Mohammed, Bassem M.; Fisher, Bernard J.; Kraskauskas, Donatas; Farkas, Daniela; Brophy, Donald F.; Fowler, Alpha A.; Natarajan, Ramesh

    2013-01-01

    Introduction: Neutrophil extracellular trap (NET) formation was recently identified as a novel mechanism to kill pathogens. However, excessive NET formation in sepsis can injure host tissues. We have recently shown that parenteral vitamin C (VitC) is protective in sepsis. Whether VitC alters NETosis is unknown. Methods: We used Gulo−/− mice as they lack the ability to synthesize VitC. Sepsis was induced by intraperitoneal infusion of a fecal stem solution (abdominal peritonitis, FIP). Some VitC deficient Gulo−/− mice received an infusion of ascorbic acid (AscA, 200 mg/kg) 30 min after induction of FIP. NETosis was assessed histologically and by quantification for circulating free DNA (cf-DNA) in serum. Autophagy, histone citrullination, endoplasmic reticulum (ER) stress, NFκB activation and apoptosis were investigated in peritoneal PMNs. Results: Sepsis produced significant NETs in the lungs of VitC deficient Gulo−/− mice and increased circulating cf-DNA. This was attenuated in the VitC sufficient Gulo−/− mice and in VitC deficient Gulo−/− mice infused with AscA. Polymorphonuclear neutrophils (PMNs) from VitC deficient Gulo−/− mice demonstrated increased activation of ER stress, autophagy, histone citrullination, and NFκB activation, while apoptosis was inhibited. VitC also significantly attenuated PMA induced NETosis in PMNs from healthy human volunteers. Conclusions: Our in vitro and in vivo findings identify VitC as a novel regulator of NET formation in sepsis. This study complements the notion that VitC is protective in sepsis settings. PMID:23939536

  9. Biogenic Iron-Rich Filaments in the Quartz Veins in the Uppermost Ediacaran Qigebulake Formation, Aksu Area, Northwestern Tarim Basin, China: Implications for Iron Oxidizers in Subseafloor Hydrothermal Systems.

    PubMed

    Zhou, Xiqiang; Chen, Daizhao; Tang, Dongjie; Dong, Shaofeng; Guo, Chuan; Guo, Zenghui; Zhang, Yanqiu

    2015-07-01

    Fe-(oxyhydr)oxide-encrusted filamentous microstructures produced by microorganisms have been widely reported in various modern and ancient extreme environments; however, the iron-dependent microorganisms preserved in hydrothermal quartz veins have not been explored in detail because of limited materials available. In this study, abundant well-preserved filamentous microstructures were observed in the hydrothermal quartz veins of the uppermost dolostones of the terminal-Ediacaran Qigebulake Formation in the Aksu area, northwestern Tarim Basin, China. These filamentous microstructures were permineralized by goethite and hematite as revealed by Raman spectroscopy and completely entombed in chalcedony and quartz cements. Microscopically, they are characterized by biogenic filamentous morphologies (commonly 20-200 μm in length and 1-5 μm in diameter) and structures (curved, tubular sheath-like, segmented, and mat-like filaments), similar to the Fe-oxidizing bacteria (FeOB) living in modern and ancient hydrothermal vent fields. A previous study revealed that quartz-barite vein swarms were subseafloor channels of low-temperature, silica-rich, diffusive hydrothermal vents in the earliest Cambrian, which contributed silica to the deposition of the overlying bedded chert of the Yurtus Formation. In this context, this study suggests that the putative filamentous FeOB preserved in the quartz veins might have thrived in the low-temperature, silica- and Fe(II)-rich hydrothermal vent channels in subseafloor mixing zones and were rapidly fossilized by subsequent higher-temperature, silica-rich hydrothermal fluids in response to waning and waxing fluctuations of diffuse hydrothermal venting. In view of the occurrence in a relatively stable passive continental margin shelf environment in Tarim Block, the silica-rich submarine hydrothermal vent system may represent a new and important geological niche favorable for FeOB colonization, which is different from their traditional

  10. Filamentation in Laser Wakefields

    NASA Astrophysics Data System (ADS)

    Los, Eva; Trines, Raoul; Silva, Luis; Bingham, Robert

    2016-10-01

    Laser filamentation instability is observed in plasma wakefields with sub-critical densities, and in high density inertial fusion plasmas. This leads to non-uniform acceleration or compression respectively. Here, we present simulation results on laser filamentation in plasma wakefields. The 2-D simulations are carried out using the particle-in-cell code Osiris. The filament intensity was found to increase exponentially before saturating. The maximum amplitude to which the highest intensity filament grew for a specific set of parameters was also recorded, and plotted against a corresponding parameter value. Clear, positively correlated linear trends were established between plasma density, transverse wavenumber k, laser pulse amplitude and maximum filament amplitude. Plasma density and maximum filament amplitude also showed a positive correlation, which saturated after a certain plasma density. Pulse duration and interaction length did not affect either filament intensity or transverse k value in a predictable manner. There was no discernible trend between pulse amplitude and filament width.

  11. Collaborative protein filaments.

    PubMed

    Ghosal, Debnath; Löwe, Jan

    2015-09-14

    It is now well established that prokaryotic cells assemble diverse proteins into dynamic cytoskeletal filaments that perform essential cellular functions. Although most of the filaments assemble on their own to form higher order structures, growing evidence suggests that there are a number of prokaryotic proteins that polymerise only in the presence of a matrix such as DNA, lipid membrane or even another filament. Matrix-assisted filament systems are frequently nucleotide dependent and cytomotive but rarely considered as part of the bacterial cytoskeleton. Here, we categorise this family of filament-forming systems as collaborative filaments and introduce a simple nomenclature. Collaborative filaments are frequent in both eukaryotes and prokaryotes and are involved in vital cellular processes including chromosome segregation, DNA repair and maintenance, gene silencing and cytokinesis to mention a few. In this review, we highlight common principles underlying collaborative filaments and correlate these with known functions.

  12. Posttranscriptional regulation of glutamine synthetase in the filamentous Cyanobacterium Anabaena sp. PCC 7120: differential expression between vegetative cells and heterocysts.

    PubMed

    Galmozzi, Carla V; Saelices, Lorena; Florencio, Francisco J; Muro-Pastor, M Isabel

    2010-09-01

    Genes homologous to those implicated in glutamine synthetase (GS) regulation by protein-protein interaction in the cyanobacterium Synechocystis sp. strain PCC 6803 are conserved in several cyanobacterial sequenced genomes. We investigated this GS regulatory mechanism in Anabaena sp. strain PCC 7120. In this strain the system operates with only one GS inactivation factor (inactivation factor 7A [IF7A]), encoded by open reading frame (ORF) asl2329 (gifA). Following addition of ammonium, expression of gifA is derepressed, leading to the synthesis of IF7A, and consequently, GS is inactivated. Upon ammonium removal, the GS activity returns to the initial level and IF7A becomes undetectable. The global nitrogen control protein NtcA binds to the gifA promoter. Constitutive high expression levels of gifA were found in an Anabaena ntcA mutant (CSE2), indicating a repressor role for NtcA. In vitro studies demonstrate that Anabaena GS is not inactivated by Synechocystis IFs (IF7 and IF17), indicating the specificity of the system. We constructed an Anabaena strain expressing a second inactivating factor, containing the amino-terminal part of IF17 from Synechocystis fused to IF7A. GS inactivation in this strain is more effective than that in the wild type (WT) and resembles that observed in Synechocystis. Finally we found differential expression of the IF system between heterocysts and vegetative cells of Anabaena.

  13. Filament Eruption Onset

    NASA Technical Reports Server (NTRS)

    Sterling, Alphonse C.; Moore, Ronald L.

    2011-01-01

    We have been investigating filament eruptions in recent years. Use filament eruptions as markers of the coronal field evolution. Data from SoHO, Yohkoh, TRACE, Hinode, and other sources. We and others have observed: (1)Filaments often show slow rise, followed by fast rise, (2) Brightenings, preflares, microflares during slow rise (3) Magnetic evolution in hours prior to eruption onset. We investigated What do Hinode and SDO show for filament eruptions?

  14. Rapid severing and motility of chloroplast-actin filaments are required for the chloroplast avoidance response in Arabidopsis.

    PubMed

    Kong, Sam-Geun; Arai, Yoshiyuki; Suetsugu, Noriyuki; Yanagida, Toshio; Wada, Masamitsu

    2013-02-01

    Phototropins (phot1 and phot2 in Arabidopsis thaliana) relay blue light intensity information to the chloroplasts, which move toward weak light (the accumulation response) and away from strong light (the avoidance response). Chloroplast-actin (cp-actin) filaments are vital for mediating these chloroplast photorelocation movements. In this report, we examine in detail the cp-actin filament dynamics by which the chloroplast avoidance response is regulated. Although stochastic dynamics of cortical actin fragments are observed on the chloroplasts, the basic mechanisms underlying the disappearance (including severing and turnover) of the cp-actin filaments are regulated differently from those of cortical actin filaments. phot2 plays a pivotal role in the strong blue light-induced severing and random motility of cp-actin filaments, processes that are therefore essential for asymmetric cp-actin formation for the avoidance response. In addition, phot2 functions in the bundling of cp-actin filaments that is induced by dark incubation. By contrast, the function of phot1 is dispensable for these responses. Our findings suggest that phot2 is the primary photoreceptor involved in the rapid reorganization of cp-actin filaments that allows chloroplasts to change direction rapidly and control the velocity of the avoidance movement according to the light's intensity and position.

  15. LMTK1 regulates dendritic formation by regulating movement of Rab11A-positive endosomes

    PubMed Central

    Takano, Tetsuya; Urushibara, Tomoki; Yoshioka, Nozomu; Saito, Taro; Fukuda, Mitsunori; Tomomura, Mineko; Hisanaga, Shin-ichi

    2014-01-01

    Neurons extend two types of neurites—axons and dendrites—that differ in structure and function. Although it is well understood that the cytoskeleton plays a pivotal role in neurite differentiation and extension, the mechanisms by which membrane components are supplied to growing axons or dendrites is largely unknown. We previously reported that the membrane supply to axons is regulated by lemur kinase 1 (LMTK1) through Rab11A-positive endosomes. Here we investigate the role of LMTK1 in dendrite formation. Down-regulation of LMTK1 increases dendrite growth and branching of cerebral cortical neurons in vitro and in vivo. LMTK1 knockout significantly enhances the prevalence, velocity, and run length of anterograde movement of Rab11A-positive endosomes to levels similar to those expressing constitutively active Rab11A-Q70L. Rab11A-positive endosome dynamics also increases in the cell body and growth cone of LMTK1-deficient neurons. Moreover, a nonphosphorylatable LMTK1 mutant (Ser34Ala, a Cdk5 phosphorylation site) dramatically promotes dendrite growth. Thus LMTK1 negatively controls dendritic formation by regulating Rab11A-positive endosomal trafficking in a Cdk5-dependent manner, indicating the Cdk5-LMTK1-Rab11A pathway as a regulatory mechanism of dendrite development as well as axon outgrowth. PMID:24672056

  16. The SOS Response Master Regulator LexA Is Associated with Sporulation, Motility and Biofilm Formation in Clostridium difficile

    PubMed Central

    Walter, Beata M.; Cartman, Stephen T.; Minton, Nigel P.; Butala, Matej; Rupnik, Maja

    2015-01-01

    The LexA regulated SOS network is a bacterial response to DNA damage of metabolic or environmental origin. In Clostridium difficile, a nosocomial pathogen causing a range of intestinal diseases, the in-silico deduced LexA network included the core SOS genes involved in the DNA repair and genes involved in various other biological functions that vary among different ribotypes. Here we describe the construction and characterization of a lexA ClosTron mutant in C. difficile R20291 strain. The mutation of lexA caused inhibition of cell division resulting in a filamentous phenotype. The lexA mutant also showed decreased sporulation, a reduction in swimming motility, greater sensitivity to metronidazole, and increased biofilm formation. Changes in the regulation of toxin A, but not toxin B, were observed in the lexA mutant in the presence of sub-inhibitory concentrations of levofloxacin. C. difficile LexA is, therefore, not only a regulator of DNA damage but also controls many biological functions associated with virulence. PMID:26682547

  17. Carvone and perillaldehyde interfere with the serum-induced formation of filamentous structures in Candida albicans at substantially lower concentrations than those causing significant inhibition of growth.

    PubMed

    McGeady, Paul; Wansley, Daniel L; Logan, David A

    2002-07-01

    Carvone and perillaldehyde were shown to inhibit the transformation of Candida albicans to a filamentous form at concentrations far lower and more biologically relevant than the concentrations necessary to inhibit growth. This morphological transformation is associated with C. albicans pathogenicity; hence these naturally occurring monoterpenes are potential lead compounds in the development of therapeutic agents against C. albicans infection.

  18. Regulation of inorganic sulfate activation in filamentous fungi. Allosteric inhibition of ATP sulfurylase by 3'-phosphoadenosine-5'-phosphosulfate.

    PubMed

    Renosto, F; Martin, R L; Wailes, L M; Daley, L A; Segel, I H

    1990-06-25

    cerevisiae are not strongly inhibited by PAPS, do not display sigmoidal initial velocity plots in the presence of PAPS, and do not contain a highly reactive cysteinyl residue whose modification induces increased [S]0.5 values and sigmoidal velocity curves. The allosteric effect of PAPS on the fungal ATP sulfurylase may be part of a sequential feedback process unique to a group of organisms that use PAPS for two diverging pathways, reductive assimilation and sulfate ester formation.

  19. Enigmatic reticulated filaments in subsurface granite.

    PubMed

    Miller, A Z; Hernández-Mariné, M; Jurado, V; Dionísio, A; Barquinha, P; Fortunato, E; Afonso, M J; Chaminé, H I; Saiz-Jimenez, C

    2012-12-01

    In the last few years, geomicrobiologists have focused their researches on the nature and origin of enigmatic reticulated filaments reported in modern and fossil samples from limestone caves and basalt lava tubes. Researchers have posed questions on these filaments concerning their nature, origin, chemistry, morphology, mode of formation and growth. A tentative microbial origin has been elusive since these filaments are found as hollow tubular sheaths and could not be affiliated to any known microorganism. We describe the presence of similar structures in a 16th century granite tunnel in Porto, Northwest Portugal. The reticulated filaments we identify exhibit fine geometry surface ornamentation formed by cross-linked Mn-rich nanofibres, surrounded by a large amount of extracellular polymeric substances. Within these Mn-rich filaments we report for the first time the occurrence of microbial cells.

  20. Neurotransmission selectively regulates synapse formation in parallel circuits in vivo.

    PubMed

    Kerschensteiner, Daniel; Morgan, Josh L; Parker, Edward D; Lewis, Renate M; Wong, Rachel O L

    2009-08-20

    Activity is thought to guide the patterning of synaptic connections in the developing nervous system. Specifically, differences in the activity of converging inputs are thought to cause the elimination of synapses from less active inputs and increase connectivity with more active inputs. Here we present findings that challenge the generality of this notion and offer a new view of the role of activity in synapse development. To imbalance neurotransmission from different sets of inputs in vivo, we generated transgenic mice in which ON but not OFF types of bipolar cells in the retina express tetanus toxin (TeNT). During development, retinal ganglion cells (RGCs) select between ON and OFF bipolar cell inputs (ON or OFF RGCs) or establish a similar number of synapses with both on separate dendritic arborizations (ON-OFF RGCs). In TeNT retinas, ON RGCs correctly selected the silenced ON bipolar cell inputs over the transmitting OFF bipolar cells, but were connected with them through fewer synapses at maturity. Time-lapse imaging revealed that this was caused by a reduced rate of synapse formation rather than an increase in synapse elimination. Similarly, TeNT-expressing ON bipolar cell axons generated fewer presynaptic active zones. The remaining active zones often recruited multiple, instead of single, synaptic ribbons. ON-OFF RGCs in TeNT mice maintained convergence of ON and OFF bipolar cells inputs and had fewer synapses on their ON arbor without changes to OFF arbor synapses. Our results reveal an unexpected and remarkably selective role for activity in circuit development in vivo, regulating synapse formation but not elimination, affecting synapse number but not dendritic or axonal patterning, and mediating independently the refinement of connections from parallel (ON and OFF) processing streams even where they converge onto the same postsynaptic cell.

  1. Membrane events in the acrosomal reaction of Limulus sperm. Membrane fusion, filament-membrane particle attachment, and the source and formation of new membrane surface

    PubMed Central

    1979-01-01

    The membranes of Limulus (horseshoe crab) sperm were examined before and during the acrosomal reaction by using the technique of freeze- fracturing and thin sectioning. We focused on three areas. First, we examined stages in the fusion of the acrosomal vacuole with the cell surface. Fusion takes place in a particle-free zone which is surrounded by a circlet of particles on the P face of the plasma membrane and an underlying circlet of particles on the P face of the acrosomal vauole membrane. These circlets of particles are present before induction. Up to nine focal points of fusion occur within the particle-free zone. Second, we describe a system of fine filaments, each 30 A in diameter, which lies between the acrosomal vacuole and the plasma membrane. These filaments change their orientation as the vacuole opens, a process that takes place in less than 50 ms. Membrane particles seen on the P face of the acrosomal vacuole membrane change their orientation at the same time and in the same way as do the filaments, thus indicating that the membrane particles and filaments are probably connected. Third, we examined the source and the point of fusion of new membrane needed to cover the acrosomal process. This new membrane is almost certainly derived from the outer nuclear envelope and appears to insert into the plasma membrane in a particle-free area adjacent to an area rich in particles. The latter is the region where the particles are probably connected to the cytoplasmic filaments. The relevance of these observations in relation to the process of fertilization of this fantastic sperm is discussed. PMID:582596

  2. Overexpression of troponin T in Drosophila muscles causes a decrease in the levels of thin-filament proteins

    PubMed Central

    2004-01-01

    Formation of the contractile apparatus in muscle cells requires co-ordinated activation of several genes and the proper assembly of their products. To investigate the role of TnT (troponin T) in the mechanisms that control and co-ordinate thin-filament formation, we generated transgenic Drosophila lines that overexpress TnT in their indirect flight muscles. All flies that overexpress TnT were unable to fly, and the loss of thin filaments themselves was coupled with ultrastructural perturbations of the sarcomere. In contrast, thick filaments remained largely unaffected. Biochemical analysis of these lines revealed that the increase in TnT levels could be detected only during the early stages of adult muscle formation and was followed by a profound decrease in the amount of this protein as well as that of other thin-filament proteins such as tropomyosin, troponin I and actin. The decrease in thin-filament proteins is not only due to degradation but also due to a decrease in their synthesis, since accumulation of their mRNA transcripts was also severely diminished. This decrease in expression levels of the distinct thin-filament components led us to postulate that any change in the amount of TnT transcripts might trigger the down-regulation of other co-regulated thin-filament components. Taken together, these results suggest the existence of a mechanism that tightly co-ordinates the expression of thin-filament genes and controls the correct stoichiometry of these proteins. We propose that the high levels of unassembled protein might act as a sensor in this process. PMID:15469415

  3. Quantifying protein diffusion and capture on filaments.

    PubMed

    Reithmann, Emanuel; Reese, Louis; Frey, Erwin

    2015-02-17

    The functional relevance of regulating proteins is often limited to specific binding sites such as the ends of microtubules or actin-filaments. A localization of proteins on these functional sites is of great importance. We present a quantitative theory for a diffusion and capture process, where proteins diffuse on a filament and stop diffusing when reaching the filament's end. It is found that end-association after one-dimensional diffusion is the main source for tip-localization of such proteins. As a consequence, diffusion and capture is highly efficient in enhancing the reaction velocity of enzymatic reactions, where proteins and filament ends are to each other as enzyme and substrate. We show that the reaction velocity can effectively be described within a Michaelis-Menten framework. Together, one-dimensional diffusion and capture beats the (three-dimensional) Smoluchowski diffusion limit for the rate of protein association to filament ends.

  4. Special issue on filamentation

    NASA Astrophysics Data System (ADS)

    Li, Ruxin; Milchberg, Howard; Mysyrowicz, André

    2014-05-01

    Journal of Physics B: Atomic, Molecular and Optical Physics is delighted to announce a forthcoming special issue on filamentation, to appear in the spring of 2015, and invites you to submit a paper. This special issue will attempt to give an overview of the present status of this field in order to create synergies and foster future developments. The issue is open to papers on the following issues: Theoretical advances on filamentation. Self-focusing and collapse. Filamentation in various media. Pulse self-compression and ultrafast processes in filaments. Molecular alignment and rotation. Filamentation tailoring. Interaction between filaments. Filament weather and pollution control. Filament induced condensation and precipitation. Terahertz science with filaments. Lasing in filaments. Filament induced molecular excitation and reaction. Electric discharge and plasma. Cross-disciplinary applications. Novel concepts related to these topics are particularly welcome. Please submit your article by 1 October 2014 (expected web publication: spring 2015) using our website http://mc04.manuscriptcentral.com/jphysb-iop. Submissions received after this date will be considered for the journal, but may not be included in the special issue. The issue will be edited by Ruxin Li, Howard Milchberg and André Mysyrowicz.

  5. Ca++-sensitizing mutations in troponin, Pi, and 2-deoxyATP alter the depressive effect of acidosis on regulated thin-filament velocity

    PubMed Central

    Longyear, Thomas J.; Turner, Matthew A.; Davis, Jonathan P.; Lopez, Joseph; Biesiadecki, Brandon

    2014-01-01

    Repeated, intense contractile activity compromises the ability of skeletal muscle to generate force and velocity, resulting in fatigue. The decrease in velocity is thought to be due, in part, to the intracellular build-up of acidosis inhibiting the function of the contractile proteins myosin and troponin; however, the underlying molecular basis of this process remains poorly understood. We sought to gain novel insight into the decrease in velocity by determining whether the depressive effect of acidosis could be altered by 1) introducing Ca++-sensitizing mutations into troponin (Tn) or 2) by agents that directly affect myosin function, including inorganic phosphate (Pi) and 2-deoxy-ATP (dATP) in an in vitro motility assay. Acidosis reduced regulated thin-filament velocity (VRTF) at both maximal and submaximal Ca++ levels in a pH-dependent manner. A truncated construct of the inhibitory subunit of Tn (TnI) and a Ca++-sensitizing mutation in the Ca++-binding subunit of Tn (TnC) increased VRTF at submaximal Ca++ under acidic conditions but had no effect on VRTF at maximal Ca++ levels. In contrast, both Pi and replacement of ATP with dATP reversed much of the acidosis-induced depression of VRTF at saturating Ca++. Interestingly, despite producing similar magnitude increases in VRTF, the combined effects of Pi and dATP were additive, suggesting different underlying mechanisms of action. These findings suggest that acidosis depresses velocity by slowing the detachment rate from actin but also by possibly slowing the attachment rate. PMID:24651988

  6. Ca++-sensitizing mutations in troponin, P(i), and 2-deoxyATP alter the depressive effect of acidosis on regulated thin-filament velocity.

    PubMed

    Longyear, Thomas J; Turner, Matthew A; Davis, Jonathan P; Lopez, Joseph; Biesiadecki, Brandon; Debold, Edward P

    2014-05-01

    Repeated, intense contractile activity compromises the ability of skeletal muscle to generate force and velocity, resulting in fatigue. The decrease in velocity is thought to be due, in part, to the intracellular build-up of acidosis inhibiting the function of the contractile proteins myosin and troponin; however, the underlying molecular basis of this process remains poorly understood. We sought to gain novel insight into the decrease in velocity by determining whether the depressive effect of acidosis could be altered by 1) introducing Ca(++)-sensitizing mutations into troponin (Tn) or 2) by agents that directly affect myosin function, including inorganic phosphate (Pi) and 2-deoxy-ATP (dATP) in an in vitro motility assay. Acidosis reduced regulated thin-filament velocity (VRTF) at both maximal and submaximal Ca(++) levels in a pH-dependent manner. A truncated construct of the inhibitory subunit of Tn (TnI) and a Ca(++)-sensitizing mutation in the Ca(++)-binding subunit of Tn (TnC) increased VRTF at submaximal Ca(++) under acidic conditions but had no effect on VRTF at maximal Ca(++) levels. In contrast, both Pi and replacement of ATP with dATP reversed much of the acidosis-induced depression of VRTF at saturating Ca(++). Interestingly, despite producing similar magnitude increases in VRTF, the combined effects of Pi and dATP were additive, suggesting different underlying mechanisms of action. These findings suggest that acidosis depresses velocity by slowing the detachment rate from actin but also by possibly slowing the attachment rate.

  7. Lin-28 regulates oogenesis and muscle formation in Drosophila melanogaster.

    PubMed

    Stratoulias, Vassilis; Heino, Tapio I; Michon, Frederic

    2014-01-01

    Understanding the control of stem cell (SC) differentiation is important to comprehend developmental processes as well as to develop clinical applications. Lin28 is a conserved molecule that is involved in SC maintenance and differentiation by regulating let-7 miRNA maturation. However, little is known about the in vivo function of Lin28. Here, we report critical roles for lin-28 during oogenesis. We found that let-7 maturation was increased in lin-28 null mutant fly ovaries. We showed that lin-28 null mutant female flies displayed reduced fecundity, due to defects in egg chamber formation. More specifically, we demonstrated that in mutant ovaries, the egg chambers fuse during early oogenesis resulting in abnormal late egg chambers. We also showed that this phenotype is the combined result of impaired germline SC differentiation and follicle SC differentiation. We suggest a model in which these multiple oogenesis defects result from a misregulation of the ecdysone signaling network, through the fine-tuning of Abrupt and Fasciclin2 expression. Our results give a better understanding of the evolutionarily conserved role of lin-28 on GSC maintenance and differentiation.

  8. Lin-28 Regulates Oogenesis and Muscle Formation in Drosophila melanogaster

    PubMed Central

    Stratoulias, Vassilis; Heino, Tapio I.; Michon, Frederic

    2014-01-01

    Understanding the control of stem cell (SC) differentiation is important to comprehend developmental processes as well as to develop clinical applications. Lin28 is a conserved molecule that is involved in SC maintenance and differentiation by regulating let-7 miRNA maturation. However, little is known about the in vivo function of Lin28. Here, we report critical roles for lin-28 during oogenesis. We found that let-7 maturation was increased in lin-28 null mutant fly ovaries. We showed that lin-28 null mutant female flies displayed reduced fecundity, due to defects in egg chamber formation. More specifically, we demonstrated that in mutant ovaries, the egg chambers fuse during early oogenesis resulting in abnormal late egg chambers. We also showed that this phenotype is the combined result of impaired germline SC differentiation and follicle SC differentiation. We suggest a model in which these multiple oogenesis defects result from a misregulation of the ecdysone signaling network, through the fine-tuning of Abrupt and Fasciclin2 expression. Our results give a better understanding of the evolutionarily conserved role of lin-28 on GSC maintenance and differentiation. PMID:24963666

  9. Radial Infall onto a Massive Molecular Filament

    NASA Astrophysics Data System (ADS)

    Battersby, Cara; Myers, Philip C.; Shirley, Yancy L.; Keto, Eric; Kirk, Helen

    The newly discovered Massive Molecular Filament (MMF) G32.02+0.05 (~ 70 pc long, 105 M⊙) has been shaped and compressed by older generations of massive stars. The similarity of this filament in physical structure (density profile, temperature) to much smaller star-forming filaments, suggests that the mechanism to form such filaments may be a universal process. The densest portion of the filament, apparent as an Infrared Dark Cloud (IRDC) shows a range of massive star formation signatures throughout. We investigate the kinematics in this filament and find widespread inverse P cygni asymmetric line profiles. These line asymmetries are interpreted as a signature of large-scale radial collapse. Using line asymmetries observed with optically thick HCO+ (1-0) and optically thin H13CO+ (1-0) across a range of massive star forming regions in the filament, we estimate the global radial infall rate of the filament to range from a few 100 to a few 1000 M⊙ Myr-1 pc-1. At its current infall rate the densest portions of the cloud will more than double their current mass within a Myr.

  10. Structural architecture of the CARMA1/Bcl10/MALT1 signalosome: nucleation-induced filamentous assembly.

    PubMed

    Qiao, Qi; Yang, Chenghua; Zheng, Chao; Fontán, Lorena; David, Liron; Yu, Xiong; Bracken, Clay; Rosen, Monica; Melnick, Ari; Egelman, Edward H; Wu, Hao

    2013-09-26

    The CARMA1/Bcl10/MALT1 (CBM) signalosome mediates antigen receptor-induced NF-κB signaling to regulate multiple lymphocyte functions. While CARMA1 and Bcl10 contain caspase recruitment domains (CARDs), MALT1 is a paracaspase with structural similarity to caspases. Here we show that the reconstituted CBM signalosome is a helical filamentous assembly in which substoichiometric CARMA1 nucleates Bcl10 filaments. Bcl10 filament formation is a highly cooperative process whose threshold is sensitized by oligomerized CARMA1 upon receptor activation. In cells, both cotransfected CARMA1/Bcl10 complex and the endogenous CBM signalosome are filamentous morphologically. Combining crystallography, nuclear magnetic resonance, and electron microscopy, we reveal the structure of the Bcl10 CARD filament and the mode of interaction between CARMA1 and Bcl10. Structure-guided mutagenesis confirmed the observed interfaces in Bcl10 filament assembly and MALT1 activation in vitro and NF-κB activation in cells. These data support a paradigm of nucleation-induced signal transduction with threshold response due to cooperativity and signal amplification by polymerization.

  11. Structural Architecture of the CARMA1/Bcl10/MALT1 Signalosome: Nucleation-Induced Filamentous Assembly

    PubMed Central

    Qiao, Qi; Yang, Chenghua; Zheng, Chao; Fontán, Lorena; David, Liron; Yu, Xiong; Bracken, Clay; Rosen, Monica; Melnick, Ari; Egelman, Edward H.; Wu, Hao

    2014-01-01

    SUMMARY The CARMA1/Bcl10/MALT1 (CBM) signalosome mediates antigen receptor-induced NF-κB signaling to regulate multiple lymphocyte functions. While CARMA1 and Bcl10 contain caspase recruitment domains (CARDs), MALT1 is a paracaspase with structural similarity to caspases. Here we show that the reconstituted CBM signalosome is a helical filamentous assembly in which substoichiometric CARMA1 nucleates Bcl10 filaments. Bcl10 filament formation is a highly cooperative process whose threshold is sensitized by oligomerized CARMA1 upon receptor activation. In cells, both cotransfected CARMA1/Bcl10 complex and the endogenous CBM signalosome are filamentous morphologically. Combining crystallography, nuclear magnetic resonance, and electron microscopy, we reveal the structure of the Bcl10 CARD filament and the mode of interaction between CARMA1 and Bcl10. Structure-guided mutagenesis confirmed the observed interfaces in Bcl10 filament assembly and MALT1 activation in vitro and NF-κB activation in cells. These data support a paradigm of nucleation-induced signal transduction with threshold response due to cooperativity and signal amplification by polymerization. PMID:24074955

  12. Cytoskeletal 10 nm filaments in cells of the algal phyla Chlorophyta, Charophyta, and Chrysophyta and their developmentally regulated and species-specific association with prosomes.

    PubMed

    Berger, S; Liddle, L B; Dillard, W L; Wittke, W; Traub, P

    2003-06-01

    10 nm diameter filaments were observed in whole-mount preparations of algae of diverse phyla: Acetabularia acetabulum and A. major (Chlorophyta), Chara australis and Nitella flexilis (Charophyta), and Poterioochromonas malhamensis (Chrysophyta). A polyclonal antibody raised against a basic, 50 kDa DNA-binding protein of A. acetabulum stains the filaments of A. acetabulumand and A. major as well as of C. australis and N. flexilis. While in the perinuclear region of A. acetabulumand and A. major and throughout the cytoplasm of P. malhamensis the 10 nm filaments have a smooth appearance, in the stalk of A. acetabulumand and A. major they are densely covered by globular structures; in C. australis and N. flexilis they are less frequently associated with such material. The morphology of a part of the globular particles is quite reminiscent of prosomes. A monoclonal antibody elicited against prosomes isolated from A. acetabulum indeed decorates the globular particles on the A. acetabulum and A. major filaments. The possible role of these filament-particle associations is discussed.

  13. An extract of Morinda citrifolia interferes with the serum-induced formation of filamentous structures in Candida albicans and inhibits germination of Aspergillus nidulans.

    PubMed

    Banerjee, Saswati; Johnson, Andrew D; Csiszar, Katalin; Wansley, Daniel L; McGeady, Paul

    2006-01-01

    An aqueous extract of Morinda citrifolia was shown to interfere with the serum-induced morphological conversion of Candida albicans from a cellular yeast to a filamentous form in vitro. The conversion of C. albicans from a cellular yeast to a filamentous form in vivo is associated with pathogenicity. No significant effect on growth in serum-free media was seen at the concentrations used to interfere with the morphological change. The same extract also inhibited the germination of Apergillus nidulans spores. These results demonstrate that M. citrifolia contains a water-soluble component or components that interfere with the morphological conversion of C. albicans and the germination of A. nidulans and may have potential therapeutic value with regard to candidiasis and aspergillosis.

  14. Supercontinuum of a 3.9 -μ m filament in air: Formation of a two-octave plateau and nonlinearly enhanced linear absorption

    NASA Astrophysics Data System (ADS)

    Panov, Nikolay A.; Shipilo, Daniil E.; Andreeva, Vera A.; Kosareva, Olga G.; Saletsky, Alexander M.; Xu, Huailiang; Polynkin, Pavel

    2016-10-01

    Through numerical simulations we reveal the scenario of 3.9 -μ m filament spectrum enrichment in the atmosphere in the cases of linear and circular polarization of the incident pulse. The discrete spectrum of odd harmonics transforms into the two-octave plateau in the case of linear polarization. In contrast, in the case of circular polarization of the incident pulse, the harmonic-free flat supercontinuum appears with the plasma onset, reaching the tenth harmonic of the input radiation. We identify the energy balance specific to the filamentation near 4 μ m : the absorption on CO2 lines in the atmosphere is accelerated by the self-phase modulation in the Kerr nonlinearity early before the plasma channel is formed. This nonlinearly enhanced linear absorption overwhelms the plasma losses and conversion of the input pulse energy to the higher harmonics as well as the plateau.

  15. Star-forming Filament Models

    NASA Astrophysics Data System (ADS)

    Myers, Philip C.

    2017-03-01

    New models of star-forming filamentary clouds are presented in order to quantify their properties and to predict their evolution. These 2D axisymmetric models describe filaments that have no core, one low-mass core, and one cluster-forming core. They are based on Plummer-like cylinders and spheroids that are bounded by a constant-density surface of finite extent. In contrast to 1D Plummer-like models, they have specific values of length and mass, they approximate observed column density maps, and their distributions of column density (N-pdfs) are pole-free. Each model can estimate the star-forming potential of a core-filament system by identifying the zone of gas dense enough to form low-mass stars and by counting the number of enclosed thermal Jeans masses. This analysis suggests that the Musca central filament may be near the start of its star-forming life, with enough dense gas to make its first ∼3 protostars, while the Coronet filament is near the midpoint of its star formation, with enough dense gas to add ∼8 protostars to its ∼20 known stars. In contrast, L43 appears to be near the end of its star-forming life, since it lacks enough dense gas to add any new protostars to the two young stellar objectsalready known.

  16. Filamentous hydrous ferric oxide biosignatures in a pipeline carrying acid mine drainage at Iron Mountain Mine, California

    USGS Publications Warehouse

    Williams, Amy J.; Alpers, Charles N.; Sumner, Dawn Y.; Campbell, Kate M.

    2017-01-01

    A pipeline carrying acidic mine effluent at Iron Mountain, CA, developed Fe(III)-rich precipitate caused by oxidation of Fe(II)aq. The native microbial community in the pipe included filamentous microbes. The pipe scale consisted of microbial filaments, and schwertmannite (ferric oxyhydroxysulfate, FOHS) mineral spheres and filaments. FOHS filaments contained central lumina with diameters similar to those of microbial filaments. FOHS filament geometry, the geochemical environment, and the presence of filamentous microbes suggest that FOHS filaments are mineralized microbial filaments. This formation of textural biosignatures provides the basis for a conceptual model for the development and preservation of biosignatures in other environments.

  17. Lamellipodin promotes actin assembly by clustering Ena/VASP proteins and tethering them to actin filaments

    PubMed Central

    Hansen, Scott D; Mullins, R Dyche

    2015-01-01

    Enabled/Vasodilator (Ena/VASP) proteins promote actin filament assembly at multiple locations, including: leading edge membranes, focal adhesions, and the surface of intracellular pathogens. One important Ena/VASP regulator is the mig-10/Lamellipodin/RIAM family of adaptors that promote lamellipod formation in fibroblasts and drive neurite outgrowth and axon guidance in neurons. To better understand how MRL proteins promote actin network formation we studied the interactions between Lamellipodin (Lpd), actin, and VASP, both in vivo and in vitro. We find that Lpd binds directly to actin filaments and that this interaction regulates its subcellular localization and enhances its effect on VASP polymerase activity. We propose that Lpd delivers Ena/VASP proteins to growing barbed ends and increases their polymerase activity by tethering them to filaments. This interaction represents one more pathway by which growing actin filaments produce positive feedback to control localization and activity of proteins that regulate their assembly. DOI: http://dx.doi.org/10.7554/eLife.06585.001 PMID:26295568

  18. 20 CFR 626.2 - Format of the Job Training Partnership Act regulations.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 20 Employees' Benefits 3 2010-04-01 2010-04-01 false Format of the Job Training Partnership Act regulations. 626.2 Section 626.2 Employees' Benefits EMPLOYMENT AND TRAINING ADMINISTRATION, DEPARTMENT OF LABOR INTRODUCTION TO THE REGULATIONS UNDER THE JOB TRAINING PARTNERSHIP ACT § 626.2 Format of the Job...

  19. 20 CFR 626.2 - Format of the Job Training Partnership Act regulations.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 20 Employees' Benefits 3 2012-04-01 2012-04-01 false Format of the Job Training Partnership Act regulations. 626.2 Section 626.2 Employees' Benefits EMPLOYMENT AND TRAINING ADMINISTRATION, DEPARTMENT OF LABOR INTRODUCTION TO THE REGULATIONS UNDER THE JOB TRAINING PARTNERSHIP ACT § 626.2 Format of the Job...

  20. 20 CFR 626.2 - Format of the Job Training Partnership Act regulations.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 20 Employees' Benefits 3 2011-04-01 2011-04-01 false Format of the Job Training Partnership Act regulations. 626.2 Section 626.2 Employees' Benefits EMPLOYMENT AND TRAINING ADMINISTRATION, DEPARTMENT OF LABOR INTRODUCTION TO THE REGULATIONS UNDER THE JOB TRAINING PARTNERSHIP ACT § 626.2 Format of the...

  1. Sequential Myosin Phosphorylation Activates Tarantula Thick Filament via a Disorder-Order Transition

    PubMed Central

    Espinoza-Fonseca, L. Michel; Alamo, Lorenzo; Pinto, Antonio; Thomas, David D.; Padrón, Raúl

    2015-01-01

    Phosphorylation of myosin regulatory light chain (RLC) N-terminal extension (NTE) activates myosin in thick filaments. RLC phosphorylation plays a primary regulatory role in smooth muscle and a secondary (modulatory) role in striated muscle, which is regulated by Ca2+ via TnC/TM on the thin filament. Tarantula striated muscle exhibits both regulatory systems: one switches on/off contraction through thin filament regulation, and another through PKC constitutively Ser35 phosphorylated swaying free heads in the thick filaments that produces quick force on twitches regulated from 0 to 50% and modulation is accomplished recruiting additional force-potentiating free and blocked heads via Ca2+4-CaM-MLCK Ser45 phosphorylation. We have used microsecond molecular dynamics (MD) simulations of tarantula RLC NTE to understand the structural basis for phosphorylation-based regulation in tarantula thick filament activation. Trajectories analysis revealed that an inter-domain salt bridges network (R39/E58,E61) facilitates formation of a stable helix-coil-helix (HCH) motif made up by helices P and A in the unphosphorylated NTE of both myosin heads. Phosphorylation of blocked head on Ser45 does not induce any substantial structural change. However, phosphorylation of free head on Ser35 disrupts this salt bridge network and induces a partial extension of helix P along RLC helix A. While not directly participating in the HCH inter-domain folding, phosphorylation of Ser35 unlocks compact structure and allows the NTE to spontaneously undergo coil-helix transitions. The modest structural change induced by subsequent Ser45 diphosphorylation monophosphorylated Ser35 free head, facilitates full helix P extension into a single structurally stable α-helix through a network of intra-domain salt bridges (pS35/R38,R39,R42). We conclude that tarantula thick filament activation is controlled by sequential Ser35-Ser45 phosphorylation via a conserved disorder-to-order transition. PMID:26038232

  2. Sequential myosin phosphorylation activates tarantula thick filament via a disorder-order transition.

    PubMed

    Espinoza-Fonseca, L Michel; Alamo, Lorenzo; Pinto, Antonio; Thomas, David D; Padrón, Raúl

    2015-08-01

    Phosphorylation of myosin regulatory light chain (RLC) N-terminal extension (NTE) activates myosin in thick filaments. RLC phosphorylation plays a primary regulatory role in smooth muscles and a secondary (modulatory) role in striated muscles, which is regulated by Ca(2+)via TnC/TM on the thin filament. Tarantula striated muscle exhibits both regulatory systems: one switches on/off contraction through thin filament regulation, and another through PKC constitutively Ser35 phosphorylated swaying free heads in the thick filaments that produces quick force on twitches regulated from 0 to 50% and modulation is accomplished recruiting additional force-potentiating free and blocked heads via Ca(2+)4-CaM-MLCK Ser45 phosphorylation. We have used microsecond molecular dynamics (MD) simulations of tarantula RLC NTE to understand the structural basis for phosphorylation-based regulation in tarantula thick filament activation. Trajectory analysis revealed that an inter-domain salt bridge network (R39/E58,E61) facilitates the formation of a stable helix-coil-helix (HCH) motif formed by helices P and A in the unphosphorylated NTE of both myosin heads. Phosphorylation of the blocked head on Ser45 does not induce any substantial structural changes. However, phosphorylation of the free head on Ser35 disrupts this salt bridge network and induces a partial extension of helix P along RLC helix A. While not directly participating in the HCH folding, phosphorylation of Ser35 unlocks a compact structure and allows the NTE to spontaneously undergo coil-helix transitions. The modest structural change induced by the subsequent Ser45 diphosphorylation monophosphorylated Ser35 free head facilitates full helix P extension into a single structurally stable α-helix through a network of intra-domain salt bridges (pS35/R38,R39,R42). We conclude that tarantula thick filament activation is controlled by sequential Ser35-Ser45 phosphorylation via a conserved disorder-to-order transition.

  3. Vimentin filament organization and stress sensing depend on its single cysteine residue and zinc binding.

    PubMed

    Pérez-Sala, Dolores; Oeste, Clara L; Martínez, Alma E; Carrasco, M Jesús; Garzón, Beatriz; Cañada, F Javier

    2015-06-02

    The vimentin filament network plays a key role in cell architecture and signalling, as well as in epithelial-mesenchymal transition. Vimentin C328 is targeted by various oxidative modifications, but its role in vimentin organization is not known. Here we show that C328 is essential for vimentin network reorganization in response to oxidants and electrophiles, and is required for optimal vimentin performance in network expansion, lysosomal distribution and aggresome formation. C328 may fulfil these roles through interaction with zinc. In vitro, micromolar zinc protects vimentin from iodoacetamide modification and elicits vimentin polymerization into optically detectable structures; in cells, zinc closely associates with vimentin and its depletion causes reversible filament disassembly. Finally, zinc transport-deficient human fibroblasts show increased vimentin solubility and susceptibility to disruption, which are restored by zinc supplementation. These results unveil a critical role of C328 in vimentin organization and open new perspectives for the regulation of intermediate filaments by zinc.

  4. The Hydrodynamic Stability of Gaseous Cosmic Filaments

    NASA Astrophysics Data System (ADS)

    Birnboim, Yuval; Padnos, Dan; Zinger, Elad

    2016-11-01

    Virial shocks at the edges of cosmic-web structures are a clear prediction of standard structure formation theories. We derive a criterion for the stability of the post-shock gas and of the virial shock itself in spherical, filamentary, and planar infall geometries. When gas cooling is important, we find that shocks become unstable, and gas flows uninterrupted toward the center of the respective halo, filament, or sheet. For filaments, we impose this criterion on self-similar infall solutions. We find that instability is expected for filament masses between 1011 and 1013 {M}⊙ Mpc-1. Using a simplified toy model, we then show that these filaments will likely feed halos with 1010 M ⊙ ≲ M halo ≲ 1013 M ⊙