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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. 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.

  3. 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.

  4. 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.

  5. 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.

  6. 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.

  7. 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

  8. 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

  9. 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

  10. 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

  11. 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.

  12. 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.

  13. 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

  14. 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

  15. 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

  16. 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.

  17. 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.

  18. 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

  19. 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

  20. 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.

  1. 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.

  2. 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.

  3. 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.

  4. NRG1, a repressor of filamentous growth in C.albicans, is down-regulated during filament induction

    PubMed Central

    Braun, Burkhard R.; Kadosh, David; Johnson, Alexander D.

    2001-01-01

    In response to a variety of external signals, the fungal pathogen Candida albicans undergoes a transition between ellipsoidal single cells (blastospores) and filaments composed of elongated cells attached end-to-end. Here we identify a DNA-binding protein, Nrg1, that represses filamentous growth in Candida probably by acting through the co-repressor Tup1. nrg1 mutant cells are predominantly filamentous under non-filament-inducing conditions and their colony morphology resembles that of tup1 mutants. We also identify two filament-specific genes, ECE1 and HWP1, whose transcription is repressed by Nrg1 under non-inducing conditions. These genes constitute a subset of those under Tup1 control, providing further evidence that Nrg1 acts by recruiting Tup1 to target genes. We show that growth in serum at 37°C, a potent inducer of filamentous growth, causes a reduction of NRG1 mRNA, suggesting that filamentous growth is induced by the down-regulation of NRG1. Consistent with this idea, expression of NRG1 from a non-regulated promoter partially blocks the induction of filamentous growth. PMID:11532939

  5. 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.

  6. 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.

  7. 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

  8. 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.

  9. 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

  10. 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

  11. 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.

  12. 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.

  13. 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.

  14. 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

  15. 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.

  16. 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

  17. 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.

  18. 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.

  19. 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.

  20. 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.

  1. 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

  2. [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.

  3. 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

  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. 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).

  6. 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.

  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. 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.

  11. 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

  12. 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.

  13. 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.

  14. 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.

  15. 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.

  16. 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.

  17. 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.

  18. 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.

  19. 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

  20. 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.

  1. 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

  2. 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).

  3. 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

  4. 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.

  5. 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.

  6. 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.

  7. 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

  8. 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

  9. 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.

  10. 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

  11. 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.

  12. 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

  13. 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.

  14. 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.

  15. 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

  16. 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

  17. 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.

  18. 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

  19. 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

  20. 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.

  1. 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.

  2. 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.

  3. 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

  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. 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

  6. 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.

  7. 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.

  8. 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

  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. 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

  11. 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

  12. 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.

  13. 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.

  14. 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

  15. 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.

  16. 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.

  17. 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

  18. 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.

  19. 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.

  20. 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.

  1. 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

  2. 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.

  3. 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.

  4. 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.

  5. 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

  6. 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.

  7. 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.

  8. 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.

  9. 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

  10. 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.

  11. 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.

  12. 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

  13. 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.

  14. 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

  15. 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.

  16. 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

  17. 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.

  18. 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

  19. 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.

  20. 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

  1. 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.

  2. 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

  3. 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.

  4. 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.

  5. 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

  6. 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.

  7. 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

  8. 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.

  9. 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

  10. 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

  11. 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.

  12. 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.

  13. 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.

  14. 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

  15. 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.

  16. 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

  17. 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.

  18. 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

  19. 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

  20. 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.

  1. 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.

  2. 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)

  3. 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

  4. 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

  5. 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.

  6. 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

  7. 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.

  8. [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.

  9. Ash1 Protein, an Asymmetrically Localized Transcriptional Regulator, Controls Filamentous Growth and Virulence of Candida albicans

    PubMed Central

    Inglis, Diane O.; Johnson, Alexander D.

    2002-01-01

    In response to a number of distinct environmental conditions, the fungal pathogen Candida albicans undergoes a morphological transition from a round, yeast form to a series of elongated, filamentous forms. This transition is believed to be critical for virulence in a mouse model of disseminated candidiasis. Here we describe the characterization of C. albicans ASH1, a gene that encodes an asymmetrically localized transcriptional regulatory protein involved in this response. We show that C. albicans ash1 mutants are defective in responding to some filament-inducing conditions. We also show that Ash1p is preferentially localized to daughter cell nuclei in the budding-yeast form of C. albicans cell growth and to the hyphal tip cells in growing filaments. Thus, Ash1p “marks” newly formed cells and presumably directs a specialized transcriptional program in these cells. Finally, we show that ASH1 is required for full virulence of C. albicans in a mouse model of disseminated candidiasis. PMID:12446785

  10. 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.

  11. 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.

  12. 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.

  13. 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.

  14. 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.

  15. 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

  16. 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.

  17. 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.

  18. 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.

  19. 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.

  20. 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.

  1. 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.

  2. 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.

  3. 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.

  4. 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.

  5. 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.

  6. 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

  7. 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.

  8. 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.

  9. 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.

  10. 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

  11. 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

  12. 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

  13. 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.

  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. 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

  16. 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

  17. 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.

  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. 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

  20. 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.

  1. 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.

  2. 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.

  3. 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.

  4. 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

  5. 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.

  6. 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.

  7. 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

  8. 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…

  9. 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.…

  10. 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.

  11. 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.

  12. 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.

  13. 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

  14. 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…

  15. 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...

  16. 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.

  17. 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.

  18. 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.

  19. 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

  20. 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.

  1. 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.

  2. 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.

  3. 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

  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. 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.

  6. 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.

  7. 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.

  8. 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

  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. 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

  11. 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

  12. 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.

  13. 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.

  14. 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

  15. 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.

  16. 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.

  17. 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.

  18. 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.

  19. 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.

  20. 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

  1. 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.

  2. 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

  3. 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

  4. 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

  5. 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.

  6. 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

  7. Contributions of EspA filaments and curli fimbriae in cellular adherence and biofilm formation of enterohemorrhagic Escherichia coli O157:H7

    Technology Transfer Automated Retrieval System (TEKTRAN)

    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...

  8. 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

  9. 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.

  10. 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

  11. 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

  12. 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.

  13. 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.

  14. 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.

  15. 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

  16. 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.

  17. 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.

  18. 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.

  19. 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.

  20. 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

  1. 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.

  2. 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.

  3. 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

  4. 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.

  5. 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.

  6. 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

  7. 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.

  8. 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.

  9. 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

  10. 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

  11. 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.

  12. 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

  13. 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

  14. 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

  15. 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.

  16. 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.

  17. 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.

  18. 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

  19. 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.

  20. 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

  1. 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

  2. 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?

  3. 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.

  4. 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

  5. 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.

  6. 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.

  7. 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

  8. 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

  9. 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.

  10. 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.

  11. 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

  12. 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.

  13. 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.

  14. 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.

  15. 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.

  16. 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.

  17. 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.

  18. 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.

  19. 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.

  20. 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.

  1. 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.

  2. 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

  3. 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

  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 ⊙ at redshift z = 3, as well as 1012 M ⊙ ≲ M halo ≲ 1015 M ⊙ at z = 0. The instability will affect the survivability of the filaments as they penetrate gaseous halos in a non-trivial way. Additionally, smaller halos accreting onto non-stable filaments will not be subject to ram pressure inside the filaments. The instreaming gas will continue toward the center and stop either once its angular momentum balances the gravitational attraction, or when its density becomes so high that it becomes self-shielded to radiation.

  5. 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...

  6. 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...

  7. 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...

  8. 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.

  9. In situ ellipsometric study of surface immobilization of flagellar filaments

    NASA Astrophysics Data System (ADS)

    Kurunczi, S.; Németh, A.; Hülber, T.; Kozma, P.; Petrik, P.; Jankovics, H.; Sebestyén, A.; Vonderviszt, F.; Fried, M.; Bársony, I.

    2010-10-01

    Protein filaments composed of thousands of subunits are promising candidates as sensing elements in biosensors. In this work in situ spectroscopic ellipsometry is applied to monitor the surface immobilization of flagellar filaments. This study is the first step towards the development of layers of filamentous receptors for sensor applications. Surface activation is performed using silanization and a subsequent glutaraldehyde crosslinking. Structure of the flagellar filament layers immobilized on activated and non-activated Si wafer substrates is determined using a two-layer effective medium model that accounted for the vertical density distribution of flagellar filaments with lengths of 300-1500 nm bound to the surface. The formation of the first interface layer can be explained by the multipoint covalent attachment of the filaments, while the second layer is mainly composed of tail pinned filaments floating upwards with the free parts. As confirmed by atomic force microscopy, covalent immobilization resulted in an increased surface density compared to absorption.

  10. Tungsten Filament Fire

    ERIC Educational Resources Information Center

    Ruiz, Michael J.; Perkins, James

    2016-01-01

    We safely remove the outer glass bulb from an incandescent lamp and burn up the tungsten filament after the glass is removed. This demonstration dramatically illustrates the necessity of a vacuum or inert gas for the environment surrounding the tungsten filament inside the bulb. Our approach has added historical importance since the incandescent…

  11. Tungsten filament fire

    NASA Astrophysics Data System (ADS)

    Ruiz, Michael J.; Perkins, James

    2016-05-01

    We safely remove the outer glass bulb from an incandescent lamp and burn up the tungsten filament after the glass is removed. This demonstration dramatically illustrates the necessity of a vacuum or inert gas for the environment surrounding the tungsten filament inside the bulb. Our approach has added historical importance since the incandescent light bulb is being replaced by compact fluorescent and LED lamps.

  12. Physical properties of cytoplasmic intermediate filaments.

    PubMed

    Block, Johanna; Schroeder, Viktor; Pawelzyk, Paul; Willenbacher, Norbert; Köster, Sarah

    2015-11-01

    Intermediate filaments (IFs) constitute a sophisticated filament system in the cytoplasm of eukaryotes. They form bundles and networks with adapted viscoelastic properties and are strongly interconnected with the other filament types, microfilaments and microtubules. IFs are cell type specific and apart from biochemical functions, they act as mechanical entities to provide stability and resilience to cells and tissues. We review the physical properties of these abundant structural proteins including both in vitro studies and cell experiments. IFs are hierarchical structures and their physical properties seem to a large part be encoded in the very specific architecture of the biopolymers. Thus, we begin our review by presenting the assembly mechanism, followed by the mechanical properties of individual filaments, network and structure formation due to electrostatic interactions, and eventually the mechanics of in vitro and cellular networks. This article is part of a Special Issue entitled: Mechanobiology.

  13. Thermal and Chemical Evolution of Collapsing Filaments

    SciTech Connect

    Gray, William J.; Scannapieco, Evan

    2013-01-15

    Intergalactic filaments form the foundation of the cosmic web that connect galaxies together, and provide an important reservoir of gas for galaxy growth and accretion. Here we present very high resolution two-dimensional simulations of the thermal and chemical evolution of such filaments, making use of a 32 species chemistry network that tracks the evolution of key molecules formed from hydrogen, oxygen, and carbon. We study the evolution of filaments over a wide range of parameters including the initial density, initial temperature, strength of the dissociating UV background, and metallicity. In low-redshift, Z ≈ 0.1Z filaments, the evolution is determined completely by the initial cooling time. If this is sufficiently short, the center of the filament always collapses to form dense, cold core containing a substantial fraction of molecules. In high-redshift, Z = 10-3Z filaments, the collapse proceeds much more slowly. This is due mostly to the lower initial temperatures, which leads to a much more modest increase in density before the atomic cooling limit is reached, making subsequent molecular cooling much less efficient. Finally, we study how the gravitational potential from a nearby dwarf galaxy affects the collapse of the filament and compare this to NGC 5253, a nearby starbusting dwarf galaxy thought to be fueled by the accretion of filament gas. In contrast to our fiducial case, a substantial density peak forms at the center of the potential. This peak evolves faster than the rest of the filament due to the increased rate at which chemical species form and cooling occur. We find that we achieve similar accretion rates as NGC 5253, but our two-dimensional simulations do not recover the formation of the giant molecular clouds that are seen in radio observations.

  14. Regulation of biofilm formation and cellular buoyancy through modulating intracellular cyclic di-GMP levels in engineered cyanobacteria.

    PubMed

    Agostoni, Marco; Waters, Christopher M; Montgomery, Beronda L

    2016-02-01

    The second messenger cyclic dimeric (3'→5') GMP (cyclic di-GMP or c-di-GMP) has been implicated in the transition between motile and sessile lifestyles in bacteria. In this study, we demonstrate that biofilm formation, cellular aggregation or flocculation, and cellular buoyancy are under the control of c-di-GMP in Synechocystis sp. PCC 6803 (Synechocystis) and Fremyella diplosiphon. Synechocystis is a unicellular cyanobacterium and displays lower levels of c-di-GMP; F. diplosiphon is filamentous and displays higher intracellular c-di-GMP levels. We transformed Synechocystis and F. diplosiphon with a plasmid for constitutive expression of genes encoding diguanylate cylase (DGC) and phosphodiesterase (PDE) proteins from Vibrio cholerae or Escherichia coli, respectively. These engineered strains allowed us to modulate intracellular c-di-GMP levels. Biofilm formation and cellular deposition were induced in the DGC-expressing Synechocystis strain which exhibited high intracellular levels of c-di-GMP; whereas strains expressing PDE in Synechocystis and F. diplosiphon to drive low intracellular levels of c-di-GMP exhibited enhanced cellular buoyancy. In addition, the PDE-expressing F. diplosiphon strain showed elevated chlorophyll levels. These results imply roles for coordinating c-di-GMP homeostasis in regulating native cyanobacterial phenotypes. Engineering exogenous DGC or PDE proteins to regulate intracellular c-di-GMP levels represents an effective tool for uncovering cryptic phenotypes or modulating phenotypes in cyanobacteria for practical applications in biotechnology applicable in photobioreactors and in green biotechnologies, such as energy-efficient harvesting of cellular biomass or the treatment of metal-containing wastewaters.

  15. A phospholipid kinase regulates actin organization and intercellular bridge formation during germline cytokinesis.

    PubMed

    Brill, J A; Hime, G R; Scharer-Schuksz, M; Fuller, M T

    2000-09-01

    The endgame of cytokinesis can follow one of two pathways depending on developmental context: resolution into separate cells or formation of a stable intercellular bridge. Here we show that the four wheel drive (fwd) gene of Drosophila melanogaster is required for intercellular bridge formation during cytokinesis in male meiosis. In fwd mutant males, contractile rings form and constrict in dividing spermatocytes, but cleavage furrows are unstable and daughter cells fuse together, producing multinucleate spermatids. fwd is shown to encode a phosphatidylinositol 4-kinase (PI 4-kinase), a member of a family of proteins that perform the first step in the synthesis of the key regulatory membrane phospholipid PIP2. Wild-type activity of the fwd PI 4-kinase is required for tyrosine phosphorylation in the cleavage furrow and for normal organization of actin filaments in the constricting contractile ring. Our results suggest a critical role for PI 4-kinases and phosphatidylinositol derivatives during the final stages of cytokinesis.

  16. Trainable Gene Regulation Networks with Applications to Drosophila Pattern Formation

    NASA Technical Reports Server (NTRS)

    Mjolsness, Eric

    2000-01-01

    This chapter will very briefly introduce and review some computational experiments in using trainable gene regulation network models to simulate and understand selected episodes in the development of the fruit fly, Drosophila melanogaster. For details the reader is referred to the papers introduced below. It will then introduce a new gene regulation network model which can describe promoter-level substructure in gene regulation. As described in chapter 2, gene regulation may be thought of as a combination of cis-acting regulation by the extended promoter of a gene (including all regulatory sequences) by way of the transcription complex, and of trans-acting regulation by the transcription factor products of other genes. If we simplify the cis-action by using a phenomenological model which can be tuned to data, such as a unit or other small portion of an artificial neural network, then the full transacting interaction between multiple genes during development can be modelled as a larger network which can again be tuned or trained to data. The larger network will in general need to have recurrent (feedback) connections since at least some real gene regulation networks do. This is the basic modeling approach taken, which describes how a set of recurrent neural networks can be used as a modeling language for multiple developmental processes including gene regulation within a single cell, cell-cell communication, and cell division. Such network models have been called "gene circuits", "gene regulation networks", or "genetic regulatory networks", sometimes without distinguishing the models from the actual modeled systems.

  17. Formation of mos RNA granules in the zebrafish oocyte that differ from cyclin B1 RNA granules in distribution, density and regulation.

    PubMed

    Horie, Mayu; Kotani, Tomoya

    2016-12-01

    Many translationally repressed mRNAs are deposited in the oocyte cytoplasm for progression of the meiotic cell cycle and early development. mos and cyclin B1 mRNAs encode proteins promoting oocyte meiosis, and translational control of these mRNAs is important for normal progression of meiotic cell division. We previously demonstrated that cyclin B1 mRNA forms RNA granules in the zebrafish and mouse oocyte cytoplasm and that the formation of RNA granules is crucial for regulating the timing of translational activation of the mRNA. However, whether the granule formation is specific to cyclin B1 mRNA remains unknown. In this study, we found that zebrafish mos mRNA forms granules distinct from those of cyclin B1 mRNA. Fluorescent in situ hybridization analysis showed that cyclin B1 RNA granules were assembled in dense clusters, while mos RNA granules were distributed diffusely in the animal polar cytoplasm. Sucrose density gradient ultracentrifugation analysis showed that the density of mos RNA granules was partly lower than that of cyclin B1 mRNA. Similar to cyclin B1 RNA granules, mos RNA granules were disassembled after initiation of oocyte maturation at the timing at which the poly(A) tail was elongated. However, while almost all of the granules of cyclin B1 were disassembled simultaneously, a fraction of mos RNA granules firstly disappeared and then a large part of them was disassembled. In addition, while cyclin B1 RNA granules were disassembled in a manner dependent on actin filament depolymerization, certain fractions of mos RNA granules were disassembled independently of actin filaments. These results suggest that cytoplasmic regulation of translationally repressed mRNAs by formation of different RNA granules is a key mechanism for translational control of distinct mRNAs in the oocyte.

  18. Environmental regulation of cloud and star formation in galactic bars

    NASA Astrophysics Data System (ADS)

    Renaud, F.; Bournaud, F.; Emsellem, E.; Agertz, O.; Athanassoula, E.; Combes, F.; Elmegreen, B.; Kraljic, K.; Motte, F.; Teyssier, R.

    2015-12-01

    The strong time-dependence of the dynamics of galactic bars yields a complex and rapidly evolving distribution of dense gas and star forming regions. Although bars mainly host regions void of any star formation activity, their extremities can gather the physical conditions for the formation of molecular complexes and mini-starbursts. Using a sub-parsec resolution hydrodynamical simulation of a Milky Way-like galaxy, we probe these conditions to explore how and where bar (hydro-)dynamics favours the formation or destruction of molecular clouds and stars. The interplay between the kpc-scale dynamics (gas flows, shear) and the parsec-scale (turbulence) is key to this problem. We find a strong dichotomy between the leading and trailing sides of the bar, in term of cloud fragmentation and in the age distribution of the young stars. After orbiting along the bar edge, these young structures slow down at the extremities of the bar, where orbital crowding increases the probability of cloud-cloud collision. We find that such events increase the Mach number of the cloud, leading to an enhanced star formation efficiency and finally the formation of massive stellar associations, in a fashion similar to galaxy-galaxy interactions. We highlight the role of bar dynamics in decoupling young stars from the clouds in which they form, and discuss the implications on the injection of feedback into the interstellar medium (ISM), in particular in the context of galaxy formation.

  19. A potential role for the COOH-terminal domain in the lateral packing of type III intermediate filaments

    PubMed Central

    1991-01-01

    To identify sites of self-association in type III intermediate filament (IF) proteins, we have taken an "anti-idiotypic antibody" approach. A mAb (anti-Ct), recognizing a similar feature near the end of the rod domain of vimentin, desmin, and peripherin (epsilon site or epsilon epitope), was characterized. Anti-idiotypic antibodies, generated by immunizing rabbits with purified anti-Ct, recognize a site (presumably "complementary" to the epsilon epitope) common among vimentin, desmin, and peripherin (beta site or beta epitope). The beta epitope is represented in a synthetic peptide (PII) modeled after the 30 COOH- terminal residues of peripherin, as seen by comparative immunoblotting assays. Consistent with the idea of an association between the epsilon and the beta site, PII binds in vitro to intact IF proteins and fragments containing the epsilon epitope, but not to IF proteins that do not react with anti-Ct. Microinjection experiments conducted in vivo and filament reconstitution assays carried out in vitro further demonstrate that "uncoupling" of this site-specific association (by competition with PII or anti-Ct) interferes with normal IF architecture, resulting in the formation of filaments and filament bundles with diameters much greater than that of the normal IFs. These thick fibers are very similar to the ones observed previously when a derivative of desmin missing 27 COOH-terminal residues was assembled in vitro (Kaufmann, E., K. Weber, and N. Geisler. 1985. J. Mol. Biol. 185:733-742). As a molecular explanation, we propose here that the epsilon and the beta sites of type III IF proteins are "complementary" and associate during filament assembly. As a result of this association, we further postulate the formation of a surface-exposed "loop" or "hairpin" structure that may sterically prevent inappropriate filament-filament aggregation and regulate filament thickness. PMID:1714461

  20. Lighting the universe with filaments.

    PubMed

    Gao, Liang; Theuns, Tom

    2007-09-14

    The first stars in the universe form when chemically pristine gas heats as it falls into dark-matter potential wells, cools radiatively because of the formation of molecular hydrogen, and becomes self-gravitating. Using supercomputer simulations, we demonstrated that the stars' properties depend critically on the currently unknown nature of the dark matter. If the dark-matter particles have intrinsic velocities that wipe out small-scale structure, then the first stars form in filaments with lengths on the order of the free-streaming scale, which can be approximately 10(20) meters (approximately 3 kiloparsecs, corresponding to a baryonic mass of approximately 10(7) solar masses) for realistic "warm dark matter" candidates. Fragmentation of the filaments forms stars with a range of masses, which may explain the observed peculiar element abundance pattern of extremely metal-poor stars, whereas coalescence of fragments and stars during the filament's ultimate collapse may seed the supermassive black holes that lurk in the centers of most massive galaxies.

  1. Mechanistic insights into the regulation of circular dorsal ruffle formation

    PubMed Central

    Itoh, Toshiki; Hasegawa, Junya

    2013-01-01

    Growth factor stimulations induce dynamic changes in the cytoskeleton beneath the plasma membrane. Among them is the formation of membrane ruffles organized in a circular array, called ‘circular dorsal ruffles’ (CDRs). Physiological functions of CDRs include downregulation of cell growth by desensitizing the signalling from growth factor receptors as well as rearrangement of adhesion sites at the onset of cell migration. For the formation of CDRs, not only the activators of actin polymerization, such as N-WASP and the Arp2/3-complex, but also membrane deforming proteins with BAR/F-BAR domains are necessary. Small GTPases are also involved in the formation of CDRs by controlling intracellular trafficking through endosomes. Moreover, recent analyses of another circular cytoskeletal structure, podosome rosettes, have revealed common molecular features shared with CDRs. Among them, the roles of PI3-kinase and phosphoinositide 5-phosphatase may hold the key to the induction of these circular structures. PMID:23175656

  2. Environmental conditions regulate the impact of plants on cloud formation

    NASA Astrophysics Data System (ADS)

    Zhao, D. F.; Buchholz, A.; Tillmann, R.; Kleist, E.; Wu, C.; Rubach, F.; Kiendler-Scharr, A.; Rudich, Y.; Wildt, J.; Mentel, Th. F.

    2017-02-01

    The terrestrial vegetation emits large amounts of volatile organic compounds (VOC) into the atmosphere, which on oxidation produce secondary organic aerosol (SOA). By acting as cloud condensation nuclei (CCN), SOA influences cloud formation and climate. In a warming climate, changes in environmental factors can cause stresses to plants, inducing changes of the emitted VOC. These can modify particle size and composition. Here we report how induced emissions eventually affect CCN activity of SOA, a key parameter in cloud formation. For boreal forest tree species, insect infestation by aphids causes additional VOC emissions which modifies SOA composition thus hygroscopicity and CCN activity. Moderate heat increases the total amount of constitutive VOC, which has a minor effect on hygroscopicity, but affects CCN activity by increasing the particles' size. The coupling of plant stresses, VOC composition and CCN activity points to an important impact of induced plant emissions on cloud formation and climate.

  3. Environmental conditions regulate the impact of plants on cloud formation

    PubMed Central

    Zhao, D. F.; Buchholz, A.; Tillmann, R.; Kleist, E.; Wu, C.; Rubach, F.; Kiendler-Scharr, A.; Rudich, Y.; Wildt, J.; Mentel, Th. F.

    2017-01-01

    The terrestrial vegetation emits large amounts of volatile organic compounds (VOC) into the atmosphere, which on oxidation produce secondary organic aerosol (SOA). By acting as cloud condensation nuclei (CCN), SOA influences cloud formation and climate. In a warming climate, changes in environmental factors can cause stresses to plants, inducing changes of the emitted VOC. These can modify particle size and composition. Here we report how induced emissions eventually affect CCN activity of SOA, a key parameter in cloud formation. For boreal forest tree species, insect infestation by aphids causes additional VOC emissions which modifies SOA composition thus hygroscopicity and CCN activity. Moderate heat increases the total amount of constitutive VOC, which has a minor effect on hygroscopicity, but affects CCN activity by increasing the particles' size. The coupling of plant stresses, VOC composition and CCN activity points to an important impact of induced plant emissions on cloud formation and climate. PMID:28218253

  4. The structures, mass motions and footpoints of solar filaments

    NASA Astrophysics Data System (ADS)

    Venkataramanasastry, Aparna

    This thesis focuses on identifying the mechanism by which solar filaments acquire mass. Some of the speculations for how a filament gets its mass are 1) injection of mass from the chromosphere into the filament structure, and 2) condensation of mass from the corona into the region of the filament channel. Mass motion at the footpoints of the filaments is studied to detect mass entering and leaving the filament body. The magnetic properties of the footpoints of the filaments are also studied. Recommendations are drawn by comparing observational properties obtained in this study with the features used in some of the previously developed models. The datasets used for this study are high-resolution image sets of centerline and Doppler wings of Halpha, obtained using the Dutch Open Telescope (DOT). The data were obtained on Oct 30, 2010. The data set contains three filaments in an active region in the northern hemisphere of the Sun. The images in each wavelength are aligned and made into movies to find the footpoints of the filaments through which the mass goes into and comes out of the filaments from and to the chromosphere, respectively. The magnetic properties of the footpoints are studied by overlaying the magnetogram images with the DOT images by using full-disk Halpha images for matching the features in the two. Of the three filaments, one of the filaments is observed to be stable throughout the duration of the observations; another filament erupts after about two hours of the beginning of observations; and the third filament is in its early stages of formation. The ends of the stable filament are clearly observed whereas the ends of the erupting filament and the forming filament are observed clearly intermittently during the duration of the observations. The animations of the region near the ends of filament 1 reveal definite injection and draining of mass via the footpoints into and out of the filament. The mass motion into and out of the filaments are observed

  5. Tracer diffusion through F-actin: effect of filament length and cross-linking.

    PubMed Central

    Jones, J D; Luby-Phelps, K

    1996-01-01

    We have determined diffusion coefficients for small (50- to 70-nm diameter) fluorescein-thiocarbamoyl-labeled Ficoll tracers through F-actin as a function of filament length and cross-linking. fx45 was used to regulate filament length and avidin/biotinylated actin or ABP-280 was used to prepare cross-linked actin gels. We found that tracer diffusion was generally independent of filament length in agreement with theoretical predictions for diffusion through solutions of rods. However, in some experiments diffusion was slower through short (< or = 1.0 micron) filaments, although this result was not consistently reproducible. Measured diffusion coefficients through unregulated F-actin and filaments of lengths > 1.0 micron were more rapid than predicted by theory for tracer diffusion through rigid, random networks, which was consistent with some degree of actin bundling. Avidin-induced cross-linking of biotinylated F-actin did not affect diffusion through unregulated F-actin, but in cases where diffusion was slower through short filaments this cross-linking method resulted in enhanced tracer diffusion rates indistinguishable from unregulated F-actin. This finding, in conjunction with increased turbidity of 1.0-micron filaments upon avidin cross-linking, indicated that this cross-linking method induces F-actin bundling. By contrast, ABP-280 cross-linking retarded diffusion through unregulated F-actin and decreased turbidity. Tracer diffusion under these conditions was well approximated by the diffusion theory. Both cross-linking procedures resulted in gel formation as determined by falling ball viscometry. These results demonstrate that network microscopic geometry is dependent on the cross-linking method, although both methods markedly increase F-actin macroscopic viscosity. PMID:8913611

  6. Distributed formation output regulation of switching heterogeneous multi-agent systems

    NASA Astrophysics Data System (ADS)

    Wang, Xiaoli

    2013-11-01

    In this article, the distributed formation output regulation problem of linear heterogeneous multi-agent systems with uncertainty under switching topology is considered. It is a generalised framework for multi-agent coordination problems, which contains or concerns a variety of important multi-agent problems in a quite unified way. Its background includes active leader following formation for the agents to maintain desired relative distances and orientations to the leader with a predefined trajectory, and multi-agent formation with environmental inputs. With the help of canonical internal model we design a distributed dynamic output feedback to handle the distributed formation output regulation problem.

  7. Filaments in the Lupus molecular clouds

    NASA Astrophysics Data System (ADS)

    Benedettini, M.; Schisano, E.; Pezzuto, S.; Elia, D.; André, P.; Könyves, V.; Schneider, N.; Tremblin, P.; Arzoumanian, D.; di Giorgio, A. M.; Di Francesco, J.; Hill, T.; Molinari, S.; Motte, F.; Nguyen-Luong, Q.; Palmeirim, P.; Rivera-Ingraham, A.; Roy, A.; Rygl, K. L. J.; Spinoglio, L.; Ward-Thompson, D.; White, G. J.

    2015-10-01

    We have studied the filaments extracted from the column density maps of the nearby Lupus 1, 3, and 4 molecular clouds, derived from photometric maps observed with the Herschel satellite. Filaments in the Lupus clouds have quite low column densities, with a median value of ˜1.5 × 1021 cm-2 and most have masses per unit length lower than the maximum critical value for radial gravitational collapse. Indeed, no evidence of filament contraction has been seen in the gas kinematics. We find that some filaments, that on average are thermally subcritical, contain dense cores that may eventually form stars. This is an indication that in the low column density regime, the critical condition for the formation of stars may be reached only locally and this condition is not a global property of the filament. Finally, in Lupus we find multiple observational evidences of the key role that the magnetic field plays in forming filaments, and determining their confinement and dynamical evolution.

  8. Snake Filament Eruption

    NASA Video Gallery

    A very long solar filament that had been snaking around the Sun erupted on Dec. 6, 2010 with a flourish. NASA's Solar Dynamics Observatory (SDO) caught the action in dramatic detail in extreme ultr...

  9. Characterization of osmotically induced filaments of Salmonella enterica.

    PubMed

    Pratt, Zachary L; Chen, Bingming; Czuprynski, Charles J; Wong, Amy C L; Kaspar, Charles W

    2012-09-01

    Salmonella enterica forms aseptate filaments with multiple nucleoids when cultured in hyperosmotic conditions. These osmotic-induced filaments are viable and form single colonies on agar plates even though they contain multiple genomes and have the potential to divide into multiple daughter cells. Introducing filaments that are formed during osmotic stress into culture conditions without additional humectants results in the formation of septa and their division into individual cells, which could present challenges to retrospective analyses of infectious dose and risk assessments. We sought to characterize the underlying mechanisms of osmotic-induced filament formation. The concentration of proteins and chromosomal DNA in filaments and control cells was similar when standardized by biomass. Furthermore, penicillin-binding proteins in the membrane of salmonellae were active in vitro. The activity of penicillin-binding protein 2 was greater in filaments than in control cells, suggesting that it may have a role in osmotic-induced filament formation. Filaments contained more ATP than did control cells in standardized cell suspensions, though the levels of two F(0)F(1)-ATP synthase subunits were reduced. Furthermore, filaments could septate and divide within 8 h in 0.2 × Luria-Bertani broth at 23°C, while nonfilamentous control cells did not replicate. Based upon the ability of filaments to septate and divide in this diluted broth, a method was developed to enumerate by plate count the number of individual, viable cells within a population of filaments. This method could aid in retrospective analyses of infectious dose of filamented salmonellae.

  10. SOLAR MAGNETIZED 'TORNADOES': RELATION TO FILAMENTS

    SciTech Connect

    Su Yang; Veronig, Astrid; Temmer, Manuela; Wang Tongjiang; Gan Weiqun

    2012-09-10

    Solar magnetized 'tornadoes', a phenomenon discovered in the solar atmosphere, appear as tornado-like structures in the corona but are rooted in the photosphere. Like other solar phenomena, solar tornadoes are a feature of magnetized plasma and therefore differ distinctly from terrestrial tornadoes. Here we report the first analysis of solar 'tornadoes' (two papers which focused on different aspects of solar tornadoes were published in the Astrophysical Journal Letters and Nature, respectively, during the revision of this Letter). A detailed case study of two events indicates that they are rotating vertical magnetic structures probably driven by underlying vortex flows in the photosphere. They usually exist as a group and are related to filaments/prominences, another important solar phenomenon whose formation and eruption are still mysteries. Solar tornadoes may play a distinct role in the supply of mass and twists to filaments. These findings could lead to a new explanation of filament formation and eruption.

  11. Arabidopsis Microtubule-Destabilizing Protein 25 Functions in Pollen Tube Growth by Severing Actin Filaments[W

    PubMed Central

    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 Ca2+, in vitro. Analysis of a mutant that bears a point mutation at the Ca2+ binding sites demonstrated that the subcellular localization of MDP25 was determined by cytosolic Ca2+ 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. PMID:24424096

  12. Regulation of Ribulose Diphosphate Formation in Vivo by Light

    PubMed Central

    Klob, W.; Kandler, O.; Tanner, W.

    1972-01-01

    Light-dependent formation of ribulose-1,5 diphosphate is completely inhibited by low concentrations of 3-(3,4-dichlorophenyl)-1,1-dimethylurea which do not severely affect cyclic photophosphorylation. Also in Scenedesmus mutant number 11, capable of cyclic photophosphorylation, cellular ribulose-1,5 diphosphate-levels do not increase upon illumination. When mutant cells are H2 adapted, however, a light-dependent formation of ribulose-1,5 diphosphate is observed in the presence of H2. From these results it has been concluded that at least part of the Calvin cycle does not operate in the dark, since a reductant is lacking which is generated in the light. PMID:16658080

  13. Platelet rich plasma promotes skeletal muscle cell migration in association with up-regulation of FAK, paxillin, and F-Actin formation.

    PubMed

    Tsai, Wen-Chung; Yu, Tung-Yang; Lin, Li-Ping; Lin, Mioa-Sui; Tsai, Ting-Ta; Pang, Jong-Hwei S

    2017-02-24

    Platelet rich plasma (PRP) contains various cytokines and growth factors which may be beneficial to the healing process of injured muscle. The aim of this study was to investigate the effect and molecular mechanism of PRP on migration of skeletal muscle cells. Skeletal muscle cells intrinsic to Sprague-Dawley rats were treated with PRP. The cell migration was evaluated by transwell filter migration assay and electric cell-substrate impedance sensing. The spreading of cells was evaluated microscopically. The formation of filamentous actin (F-actin) cytoskeleton was assessed by immunofluorescence staining. The protein expressions of paxillin and focal adhesion kinase (FAK) were assessed by Western blot analysis. Transfection of paxillin small-interfering RNA (siRNAs) to muscle cells was performed to validate the role of paxillin in PRP-mediated promotion of cell migration. Dose-dependently PRP promotes migration of and spreading and muscle cells. Protein expressions of paxillin and FAK were up-regulated dose-dependently. F-actin formation was also enhanced by PRP treatment. Furthermore, the knockdown of paxillin expression impaired the effect of PRP to promote cell migration. It was concluded that PRP promoting migration of muscle cells is associated with up-regulation of proteins expression of paxillin and FAK as well as increasing F-actin formation. © 2017 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res.

  14. Effects of troponin T cardiomyopathy mutations on the calcium sensitivity of the regulated thin filament and the actomyosin cross-bridge kinetics of human β-cardiac myosin.

    PubMed

    Sommese, Ruth F; Nag, Suman; Sutton, Shirley; Miller, Susan M; Spudich, James A; Ruppel, Kathleen M

    2013-01-01

    Hypertrophic cardiomyopathy (HCM) and dilated cardiomyopathy (DCM) lead to significant cardiovascular morbidity and mortality worldwide. Mutations in the genes encoding the sarcomere, the force-generating unit in the cardiomyocyte, cause familial forms of both HCM and DCM. This study examines two HCM-causing (I79N, E163K) and two DCM-causing (R141W, R173W) mutations in the troponin T subunit of the troponin complex using human β-cardiac myosin. Unlike earlier reports using various myosin constructs, we found that none of these mutations affect the maximal sliding velocities or maximal Ca(2+)-activated ADP release rates involving the thin filament human β-cardiac myosin complex. Changes in Ca(2+) sensitivity using the human myosin isoform do, however, mimic changes seen previously with non-human myosin isoforms. Transient kinetic measurements show that these mutations alter the kinetics of Ca(2+) induced conformational changes in the regulatory thin filament proteins. These changes in calcium sensitivity are independent of active, cycling human β-cardiac myosin.

  15. Effects of Troponin T Cardiomyopathy Mutations on the Calcium Sensitivity of the Regulated Thin Filament and the Actomyosin Cross-Bridge Kinetics of Human β-Cardiac Myosin

    PubMed Central

    Sutton, Shirley; Miller, Susan M.; Spudich, James A.; Ruppel, Kathleen M.

    2013-01-01

    Hypertrophic cardiomyopathy (HCM) and dilated cardiomyopathy (DCM) lead to significant cardiovascular morbidity and mortality worldwide. Mutations in the genes encoding the sarcomere, the force-generating unit in the cardiomyocyte, cause familial forms of both HCM and DCM. This study examines two HCM-causing (I79N, E163K) and two DCM-causing (R141W, R173W) mutations in the troponin T subunit of the troponin complex using human β-cardiac myosin. Unlike earlier reports using various myosin constructs, we found that none of these mutations affect the maximal sliding velocities or maximal Ca2+-activated ADP release rates involving the thin filament human β-cardiac myosin complex. Changes in Ca2+ sensitivity using the human myosin isoform do, however, mimic changes seen previously with non-human myosin isoforms. Transient kinetic measurements show that these mutations alter the kinetics of Ca2+ induced conformational changes in the regulatory thin filament proteins. These changes in calcium sensitivity are independent of active, cycling human β-cardiac myosin. PMID:24367593

  16. Redox-driven conductance switching via filament formation and dissolution in carbon/molecule/TiO2/Ag molecular electronic junctions.

    PubMed

    Ssenyange, Solomon; Yan, Haijun; McCreery, Richard L

    2006-12-05

    Carbon/molecule/TiO2/Au molecular electronic junctions show robust conductance switching, in which a metastable high conductance state may be induced by a voltage pulse which results in redox reactions in the molecular and TiO2 layers. When Ag is substituted for Au as the "top contact", dramatically different current/voltage curves and switching behavior result. When the carbon substrate is biased negative, an apparent breakdown occurs, leading to a high conductance state which is stable for at least several hours. Upon scanning to positive bias, the conductance returns to a low state, and the cycle may be repeated hundreds of times. Similar effects are observed when Cu is substituted for Au and for three different molecular layers as well as "control" junctions of the type carbon/TiO2/Ag/Au. The polarity of the "switching" is reversed when the Ag layer is between the carbon and molecular layers, and the conductance change is suppressed at low temperature. Pulse experiments show very erratic transitions between high and low conductivity states, particularly near the switching threshold. The results are consistent with a switching mechanism based on Ag or Cu oxidation, transport of their ions through the TiO2, and reduction at the carbon to form a metal filament.

  17. Phosphoinositide 3-kinase p85beta regulates invadopodium formation

    PubMed Central

    Cariaga-Martínez, Ariel E.; Cortés, Isabel; García, Esther; Pérez-García, Vicente; Pajares, María J.; Idoate, Miguel A.; Redondo-Muñóz, Javier; Antón, Inés M.; Carrera, Ana C.

    2014-01-01

    ABSTRACT The acquisition of invasiveness is characteristic of tumor progression. Numerous genetic changes are associated with metastasis, but the mechanism by which a cell becomes invasive remains unclear. Expression of p85β, a regulatory subunit of phosphoinositide-3-kinase, markedly increases in advanced carcinoma, but its mode of action is unknown. We postulated that p85β might facilitate cell invasion. We show that p85β localized at cell adhesions in complex with focal adhesion kinase and enhanced stability and maturation of cell adhesions. In addition, p85β induced development at cell adhesions of an F-actin core that extended several microns into the cell z-axis resembling the skeleton of invadopodia. p85β lead to F-actin polymerization at cell adhesions by recruiting active Cdc42/Rac at these structures. In accordance with p85β function in invadopodium-like formation, p85β levels increased in metastatic melanoma and p85β depletion reduced invadopodium formation and invasion. These results show that p85β enhances invasion by inducing cell adhesion development into invadopodia-like structures explaining the metastatic potential of tumors with increased p85β levels. PMID:25217619

  18. Phosphorylation of K+ channels at single residues regulates memory formation

    PubMed Central

    Vernon, Jeffrey; Irvine, Elaine E.; Peters, Marco; Jeyabalan, Jeshmi

    2016-01-01

    Phosphorylation is a ubiquitous post-translational modification of proteins, and a known physiological regulator of K+ channel function. Phosphorylation of K+ channels by kinases has long been presumed to regulate neuronal processing and behavior. Although circumstantial evidence has accumulated from behavioral studies of vertebrates and invertebrates, the contribution to memory of single phosphorylation sites on K+ channels has never been reported. We have used gene targeting in mice to inactivate protein kinase A substrate residues in the fast-inactivating subunit Kv4.2 (T38A mutants), and in the small-conductance Ca2+-activated subunit SK1 (S105A mutants). Both manipulations perturbed a specific form of memory, leaving others intact. T38A mutants had enhanced spatial memory for at least 4 wk after training, whereas performance in three tests of fear memory was unaffected. S105A mutants were impaired in passive avoidance memory, sparing fear, and spatial memory. Together with recent findings that excitability governs the participation of neurons in a memory circuit, this result suggests that the memory type supported by neurons may depend critically on the phosphorylation of specific K+ channels at single residues. PMID:26980786

  19. Profilin1 Regulates Sternum Development and Endochondral Bone Formation

    PubMed Central

    Miyajima, Daisuke; Hayata, Tadayoshi; Suzuki, Takafumi; Hemmi, Hiroaki; Nakamoto, Tetsuya; Notomi, Takuya; Amagasa, Teruo; Böttcher, Ralph T.; Costell, Mercedes; Fässler, Reinhard; Ezura, Yoichi; Noda, Masaki

    2012-01-01

    Bone development is a dynamic process that requires cell motility and morphological adaptation under the control of actin cytoskeleton. This actin cytoskeleton system is regulated by critical modulators including actin-binding proteins. Among them, profilin1 (Pfn1) is a key player to control actin fiber structure, and it is involved in a number of cellular activities such as migration. During the early phase of body development, skeletal stem cells and osteoblastic progenitor cells migrate to form initial rudiments for future skeletons. During this migration, these cells extend their process based on actin cytoskeletal rearrangement to locate themselves in an appropriate location within microenvironment. However, the role of Pfn1 in regulation of mesenchymal progenitor cells (MPCs) during skeletal development is incompletely understood. Here we examined the role of Pfn1 in skeletal development using a genetic ablation of Pfn1 in MPCs by using Prx1-Cre recombinase. We found that Pfn1 deficiency in MPCs caused complete cleft sternum. Notably, Pfn1-deficient mice exhibited an absence of trabecular bone in the marrow space of appendicular long bone. This phenotype is location-specific, as Pfn1 deficiency did not largely affect osteoblasts in cortical bone. Pfn1 deficiency also suppressed longitudinal growth of long bone. In vitro, Pfn1 deficiency induced retardation of osteoblastic cell migration. These observations revealed that Pfn1 is a critical molecule for the skeletal development, and this could be at least in part associated with the retardation of cell migration PMID:22773831

  20. Insight into Skin Tumorigenesis Highlighting the Function of Epigenetic Regulators in SCC Formation

    DTIC Science & Technology

    2013-10-01

    Epigenetic regulation of cellular memory by the polycomb and trithorax group proteins . Annu Rev Genet 2004;38:413-443...lys 27. Genes Dev 2003;17:1823-1828. 9 Francis NJ, Kingston RE, Woodcock CL: Chromatin compaction by a polycomb group protein complex. Science...Highlighting the Function of Epigenetic Regulators in SCC Formation PRINCIPAL INVESTIGATOR: Jisheng Zhang CONTRACTING

  1. Signaling pathways regulating cartilage growth plate formation and activity.

    PubMed

    Samsa, William E; Zhou, Xin; Zhou, Guang

    2017-02-01

    The growth plate is a highly specialized and dynamic cartilage structure that serves many essential functions in skeleton patterning, growth and endochondral ossification in developing vertebrates. Major signaling pathways initiated by classical morphogens and by other systemic and tissue-specific factors are intimately involved in key aspects of growth plate development. As a corollary of these essential functions, disturbances in these pathways due to mutations or environmental factors lead to severe skeleton disorders. Here, we review these pathways and the most recent progress made in understanding their roles in chondrocyte differentiation in growth plate development and activity. Furthermore, we discuss newly uncovered pathways involved in growth plate formation, including mTOR, the circadian clock, and the COP9 signalosome.

  2. The role of ClpP, RpoS and CsrA in growth and filament formation of Salmonella enterica serovar Typhimurium at low temperature

    PubMed Central

    2014-01-01

    Background Salmonellae are food-borne pathogens of great health and economic importance. To pose a threat to humans, Salmonellae normally have to cope with a series of stressful conditions in the food chain, including low temperature. In the current study, we evaluated the importance of the Clp proteolytic complex and the carbon starvation protein, CsrA, for the ability of Salmonella Typhimurium to grow at low temperature. Results A clpP mutant was severely affected in growth and formed pin point colonies at 10°C. Contrary to this, rpoS and clpP/rpoS mutants were only slightly affected. The clpP mutant formed cold resistant suppressor mutants at a frequency of 2.5 × 10−3 and these were found not to express RpoS. Together these results indicated that the impaired growth of the clpP mutant was caused by high level of RpoS. Evaluation by microscopy of the clpP mutant revealed that it formed filamentous cells when grown at 10°C, and this phenotype too, disappered when rpoS was mutated in parallel indicating a RpoS-dependency. A csrA (sup) mutant was also growth attenuated a low temperature. An rpoS/csrA (sup) double mutant was also growth attenuated, indicating that the phenotype of the csrA mutant was independent from RpoS. Conclusions The cold sensitivity of clpP mutant was associated with increased levels of RpoS and probably caused by toxic levels of RpoS. Although a csrA mutant also accumulated high level of RpoS, growth impairment caused by lack of csrA was not related to RpoS levels in a similar way. PMID:25123657

  3. Characterization of HI Filaments

    NASA Astrophysics Data System (ADS)

    Lubar, Emily; Verschuur, Gerrit L.

    2017-01-01

    We characterized the properties of dramatic interstellar HI filaments to learn more about the dynamics and structure of such features. Using Gauss fitting software, we searched the Effelsburg-Bonn HI Survey data for indications of a simple twisting (toroidal) motion across these filaments. Instead, we found that the structure was more complicated than expected. Apparent angular widths of several filaments were measured using the Galactic Arecibo L-band Feed Array HI (GALFA-HI), Bonn, and Leident/Argentine/Bonn (LAB) surveys. Based on filament widths and other parameters, we conclude that magnetism is the dominant force opposing internal motion and maintaining the structure of these filaments. The apparent width as a function of beam width closely follows a relationship reported in 1993 for HI features in general. They tend to subtend an angle two times the beam width, suggesting that the features remain unresolved.The Arecibo Observatory is operated by SRI International under a cooperative agreement with the National Science Foundation (AST-1100968), and in alliance with Ana G. Méndez-Universidad Metropolitana, and the Universities Space Research Association. The Arecibo Observatory REU is funded under grant AST-1559849 to Universidad Metropolitana.

  4. Physical principles of filamentous protein self-assembly kinetics

    NASA Astrophysics Data System (ADS)

    Michaels, Thomas C. T.; Liu, Lucie X.; Meisl, Georg; Knowles, Tuomas P. J.

    2017-04-01

    The polymerization of proteins and peptides into filamentous supramolecular structures is an elementary form of self-organization of key importance to the functioning biological systems, as in the case of actin biofilaments that compose the cellular cytoskeleton. Aberrant filamentous protein self-assembly, however, is associated with undesired effects and severe clinical disorders, such as Alzheimer’s and Parkinson’s diseases, which, at the molecular level, are associated with the formation of certain forms of filamentous protein aggregates known as amyloids. Moreover, due to their unique physicochemical properties, protein filaments are finding extensive applications as biomaterials for nanotechnology. With all these different factors at play, the field of filamentous protein self-assembly has experienced tremendous activity in recent years. A key question in this area has been to elucidate the microscopic mechanisms through which filamentous aggregates emerge from dispersed proteins with the goal of uncovering the underlying physical principles. With the latest developments in the mathematical modeling of protein aggregation kinetics as well as the improvement of the available experimental techniques it is now possible to tackle many of these complex systems and carry out detailed analyses of the underlying microscopic steps involved in protein filament formation. In this paper, we review some classical and modern kinetic theories of protein filament formation, highlighting their use as a general strategy for quantifying the molecular-level mechanisms and transition states involved in these processes.

  5. Physical principles of filamentous protein self-assembly kinetics.

    PubMed

    Michaels, Thomas; Liu, Xiaoxuan; Meisl, Georg; Knowles, Tuomas P J

    2017-02-07

    The polymerization of proteins and peptides into filamentous supramolecular structures is an elementary form of self-organization of key importance to the functioning biological systems, as in the case of actin biofilaments that compose the cellular cytoskeleton. Aberrant filamentous protein self-assembly, however, is associated with undesired effects and severe clinical disorders, such as Alzheimer's and Parkinson's diseases, which, at the molecular level, are associated with the formation of certain forms of filamentous protein aggregates known as amyloids. Moreover, due to their unique physicochemical properties, protein filaments are finding extensive applications as biomaterials for nanotechnology. With all these different factors at play, the field of filamentous protein self-assembly has experienced tremendous activity in recent years. A key question in this area has been to elucidate the microscopic mechanisms through which filamentous aggregates emerge from dispersed proteins with the goal of uncovering the underlying physical principles. With the latest developments in the mathematical modeling of protein aggregation kinetics as well as the improvement of the available experimental techniques it is now possible to tackle many of these complex systems and carry out detailed analyses of the underlying microscopic steps involved in protein filament formation. In this paper, we review some classical and modern kinetic theories of protein filament formation, highlighting their use as a general strategy for quantifying the molecular-level mechanisms and transition states involved in these processes.

  6. Subunit positioning and stator filament stiffness in regulation and power transmission in the V1 motor of the Manduca sexta V-ATPase.

    PubMed

    Muench, Stephen P; Scheres, Sjors H W; Huss, Markus; Phillips, Clair; Vitavska, Olga; Wieczorek, Helmut; Trinick, John; Harrison, Michael A

    2014-01-23

    The vacuolar H(+)-ATPase (V-ATPase) is an ATP-driven proton pump essential to the function of eukaryotic cells. Its cytoplasmic V1 domain is an ATPase, normally coupled to membrane-bound proton pump Vo via a rotary mechanism. How these asymmetric motors are coupled remains poorly understood. Low energy status can trigger release of V1 from the membrane and curtail ATP hydrolysis. To investigate the molecular basis for these processes, we have carried out cryo-electron microscopy three-dimensional reconstruction of deactivated V1 from Manduca sexta. In the resulting model, three peripheral stalks that are parts of the mechanical stator of the V-ATPase are clearly resolved as unsupported filaments in the same conformations as in the holoenzyme. They are likely therefore to have inherent stiffness consistent with a role as flexible rods in buffering elastic power transmission between the domains of the V-ATPase. Inactivated V1 adopted a homogeneous resting state with one open active site adjacent to the stator filament normally linked to the H subunit. Although present at 1:1 stoichiometry with V1, both recombinant subunit C reconstituted with V1 and its endogenous subunit H were poorly resolved in three-dimensional reconstructions, suggesting structural heterogeneity in the region at the base of V1 that could indicate positional variability. If the position of H can vary, existing mechanistic models of deactivation in which it binds to and locks the axle of the V-ATPase rotary motor would need to be re-evaluated.

  7. Surface microcracks signal osteoblasts to regulate alignment and bone formation

    PubMed Central

    Shu, Yutian; Baumann, Melissa J.; Case, Eldon D.; Irwin, Regina K.; Meyer, Sarah E.; Pearson, Craig S.; McCabe, Laura R.

    2014-01-01

    Microcracks are present in bone and can result from fatigue damage due to repeated, cyclically applied stresses. From a mechanical point, microcracks can dissipate strain energy at the advancing tip of a crack to improve overall bone toughness. Physiologically, microcracks are thought to trigger bone remodeling. Here, we examine the effect of microcracks specifically on osteoblasts, which are bone-forming cells, by comparing cell responses on microcracked versus non-microcracked hydroxyapatite (HA) specimens. Osteoblast attachment was found to be greater on microcracked HA specimens (p<0.05). More importantly, we identified the preferential alignment of osteoblasts in the direction of the microcracks on HA. Cells also displayed a preferential attachment that was 75 to 90 μm away from the microcrack indent. After 21 days of culture, osteoblast maturation was notably enhanced on the HA with microcracks, as indicated by increased alkaline phosphatase activity and gene expression. Furthermore, examination of bone deposition by confocal laser scanning microscope indicated preferential mineralization at microcrack indentation sites. Dissolution studies indicate that the microcracks increase calcium release, which could contribute to osteoblast responses. Our findings suggest that microcracks signal osteoblast attachment and bone formation/healing. PMID:25280696

  8. Dscam-Mediated Cell Recognition Regulates Neural Circuit Formation

    PubMed Central

    Hattori, Daisuke; Millard, S. Sean; Wojtowicz, Woj M.; Zipursky, S. Lawrence

    2009-01-01

    The Dscam family of immunoglobulin cell surface proteins mediates recognition events between neurons that play an essential role in the establishment of neural circuits. The Drosophila Dscam1 locus encodes tens of thousands of cell surface proteins via alternative splicing. These isoforms exhibit exquisite isoform-specific binding in vitro that mediates homophilic repulsion in vivo. These properties provide the molecular basis for self-avoidance, an essential developmental mechanism that allows axonal and dendritic processes to uniformly cover their synaptic fields. In a mechanistically similar fashion, homophilic repulsion mediated by Drosophila Dscam2 prevents processes from the same class of cells from occupying overlapping synaptic fields through a process called tiling. Genetic studies in the mouse visual system support the view that vertebrate DSCAM also promotes both self-avoidance and tiling. By contrast, DSCAM and DSCAM-L promote layer-specific targeting in the chick visual system, presumably through promoting homophilic adhesion. The fly and mouse studies underscore the importance of homophilic repulsion in regulating neural circuit assembly, whereas the chick studies suggest that DSCA Mproteins may mediate a variety of different recognition events during wiring in a context-dependent fashion. PMID:18837673

  9. Thermodynamic model of heterochromatin formation through epigenetic regulation

    NASA Astrophysics Data System (ADS)

    Mulligan, Peter J.; Koslover, Elena F.; Spakowitz, Andrew J.

    2015-02-01

    Gene regulation in eukaryotes requires the segregation of silenced genomic regions into densely packed heterochromatin, leaving the active genes in euchromatin regions more accessible. We introduce a model that connects the presence of epigenetically inherited histone marks, methylation at histone 3 lysine-9, to the physical compaction of chromatin fibers via the binding of heterochromatin protein 1 (HP1). Our model demonstrates some of the key physical features that are necessary to explain experimental observations. In particular, we demonstrate that strong cooperative interactions among the HP1 proteins are necessary to see the phase segregation of heterochromatin and euchromatin regions. We also explore how the cell can use the concentration of HP1 to control condensation and under what circumstances there is a threshold of methylation over which the fibers will compact. Finally, we consider how different potential in vivo fiber structures as well as the flexibility of the histone 3 tail can affect the bridging of HP1. Many of the observations that we make about the HP1 system are guided by general thermodynamics principles and thus could play a role in other DNA organizational processes such as the binding of linker histones.

  10. Magnetically Self-regulated Formation of Early Protoplanetary Disks

    NASA Astrophysics Data System (ADS)

    Hennebelle, Patrick; Commerçon, Benoît; Chabrier, Gilles; Marchand, Pierre

    2016-10-01

    The formation of protoplanetary disks during the collapse of molecular dense cores is significantly influenced by angular momentum transport, notably by the magnetic torque. In turn, the evolution of the magnetic field is determined by dynamical processes and non-ideal MHD effects such as ambipolar diffusion. Considering simple relations between various timescales characteristic of the magnetized collapse, we derive an expression for the early disk radius, r≃ 18 {au} {({η }{AD}/0.1{{s}})}2/9{({B}z/0.1{{G}})}-4/9{(M/0.1{M}⊙ )}1/3, where M is the total disk plus protostar mass, {η }{AD} is the ambipolar diffusion coefficient, and B z is the magnetic field in the inner part of the core. This is significantly smaller than the disks that would form if angular momentum was conserved. The analytical predictions are confronted against a large sample of 3D, non-ideal MHD collapse calculations covering variations of a factor 100 in core mass, a factor 10 in the level of turbulence, a factor 5 in rotation, and magnetic mass-to-flux over critical mass-to-flux ratios 2 and 5. The disk radius estimates are found to agree with the numerical simulations within less than a factor 2. A striking prediction of our analysis is the weak dependence of circumstellar disk radii upon the various relevant quantities, suggesting weak variations among class-0 disk sizes. In some cases, we note the onset of large spiral arms beyond this radius.

  11. Pathway and regulation of erythritol formation in Leuconostoc oenos.

    PubMed Central

    Veiga-da-Cunha, M; Santos, H; Van Schaftingen, E

    1993-01-01

    It was recently observed that Leuconostoc oenos GM, a wine lactic acid bacterium, produced erythritol anaerobically from glucose but not from fructose or ribose and that this production was almost absent in the presence of O2. In this study, the pathway of formation of erythritol from glucose in L. oenos was shown to involve the isomerization of glucose 6-phosphate to fructose 6-phosphate by a phosphoglucose isomerase, the cleavage of fructose 6-phosphate by a phosphoketolase, the reduction of erythrose 4-phosphate by an erythritol 4-phosphate dehydrogenase and, finally, the hydrolysis of erythritol 4-phosphate to erythritol by a phosphatase. Fructose 6-phosphate phosphoketolase was copurified with xylulose 5-phosphate phosphoketolase, and the activity of the latter was competitively inhibited by fructose 6-phosphate, with a Ki of 26 mM, corresponding to the Km of fructose 6-phosphate phosphoketolase (22 mM). These results suggest that the two phosphoketolase activities are borne by a single enzyme. Extracts of L. oenos were also found to contain NAD(P)H oxidase, which must be largely responsible for the reoxidation of NADPH and NADH in cells incubated in the presence of O2. In cells incubated with glucose, the concentrations of glucose 6-phosphate and of fructose 6-phosphate were higher in the absence of O2 than in its presence, explaining the stimulation by anaerobiosis of erythritol production. The increase in the hexose 6-phosphate concentration is presumably the result of a functional inhibition of glucose 6-phosphate dehydrogenase because of a reduction in the availability of NADP. PMID:8391532

  12. Pressure regulates osteoclast formation and MCSF expression in marrow culture.

    PubMed

    Rubin, J; Biskobing, D; Fan, X; Rubin, C; McLeod, K; Taylor, W R

    1997-01-01

    One of the forces generated during skeletal loading is hydrostatic pressure. In the work presented here, the ability of increased pressure to influence recruitment of osteoclasts was evaluated. Murine marrow cultures, with pO2 and pCO2 kept constant, were subjected to either control (1.0 atm) or elevated (1.37 or 2.0 atm) hydrostatic pressure. As compared to control, cultures pressurized for 6 days at 1.37 atm formed less osteoclast-like cells (OCLC) (71 +/- 6% of control, P < 0.0001). A similar degree of inhibition occurred in cultures exposed to pressure during days 2-4 only (62 +/- 6%), while treatment during days 5-7 failed to inhibit the OCLC number relative to control (99 +/- 5%). Delivery of 2.0 atm pressure on days 2-4 generated 52 +/- 4% OCLC compared to control. Since macrophage colony stimulating factor (MCSF)-dependent proliferation of osteoclast precursors occurs during the pressure-sensitive period, semiquantitative RT-PCR for MCSF mRNA was performed after 3 days in 1.37 atm (days 2-4). As compared to controls, pressure caused a decrease in mRNA coding for the membrane bound form of MCSF (71.2 +/- 4% (n = 25, P < or = 0.05), while the MCSF RT-PCR product representing the secreted form showed no consistent change. This lack of response of the soluble MCSF RT-PCR product was expected, as levels of bioassayable MCSF were not altered by pressure. Extrapolating these data to in vivo conditions suggests that load-bearing will inhibit the formation of osteoclasts.

  13. Aerogel-supported filament

    DOEpatents

    Wuest, C.R.; Tillotson, T.M.; Johnson, C.V. III

    1995-05-16

    The present invention is a thin filament embedded in a low density aerogel for use in radiation detection instruments and incandescent lamps. The aerogel provides a supportive matrix that is thermally and electrically nonconductive, mechanically strong, highly porous, gas-permeable, and transparent to ionizing radiation over short distances. A low density, open-cell aerogel is cast around a fine filament or wire, which allows the wire to be positioned with little or no tension and keeps the wire in place in the event of breakage. The aerogel support reduces the stresses on the wire caused by vibrational, gravitational, electrical, and mechanical forces. 6 Figs.

  14. Aerogel-supported filament

    DOEpatents

    Wuest, Craig R.; Tillotson, Thomas M.; Johnson, III, Coleman V.

    1995-01-01

    The present invention is a thin filament embedded in a low density aerogel for use in radiation detection instruments and incandescent lamps. The aerogel provides a supportive matrix that is thermally and electrically nonconductive, mechanically strong, highly porous, gas-permeable, and transparent to ionizing radiation over short distances. A low density, open-cell aerogel is cast around a fine filament or wire, which allows the wire to be positioned with little or no tension and keeps the wire in place in the event of breakage. The aerogel support reduces the stresses on the wire caused by vibrational, gravitational, electrical, and mechanical forces.

  15. Staurosporine Induces Filamentation in the Human Fungal Pathogen Candida albicans via Signaling through Cyr1 and Protein Kinase A.

    PubMed

    Xie, Jinglin L; O'Meara, Teresa R; Polvi, Elizabeth J; Robbins, Nicole; Cowen, Leah E

    2017-01-01

    Protein kinases are key regulators of signal transduction pathways that participate in diverse cellular processes. In fungal pathogens, kinases regulate signaling pathways that govern drug resistance, stress adaptation, and pathogenesis. The impact of kinases on the fungal regulatory circuitry has recently garnered considerable attention in the opportunistic fungal pathogen Candida albicans, which is a leading cause of human morbidity and mortality. Complex regulatory circuitry governs the C. albicans morphogenetic transition between yeast and filamentous growth, which is a key virulence trait. Here, we report that staurosporine, a promiscuous kinase inhibitor that abrogates fungal drug resistance, also influences C. albicans morphogenesis by inducing filamentation in the absence of any other inducing cue. We further establish that staurosporine exerts its effect via the adenylyl cyclase Cyr1 and the cyclic AMP (cAMP)-dependent protein kinase A (PKA). Strikingly, filamentation induced by staurosporine does not require the known upstream regulators of Cyr1, Ras1 or Pkc1, or effectors downstream of PKA, including Efg1. We further demonstrate that Cyr1 is capable of activating PKA to enable filamentation in response to staurosporine through a mechanism that does not require degradation of the transcriptional repressor Nrg1. We establish that staurosporine-induced filamentation is accompanied by a defect in septin ring formation, implicating cell cycle kinases as potential staurosporine targets underpinning this cellular response. Thus, we establish staurosporine as a chemical probe to elucidate the architecture of cellular signaling governing fungal morphogenesis and highlight the existence of novel circuitry through which the Cyr1 and PKA govern a key virulence trait. IMPORTANCE The impact of fungal pathogens on human health is devastating. One of the most pervasive fungal pathogens is Candida albicans, which kills ~40% of people suffering from bloodstream

  16. Staurosporine Induces Filamentation in the Human Fungal Pathogen Candida albicans via Signaling through Cyr1 and Protein Kinase A

    PubMed Central

    Xie, Jinglin L.; O’Meara, Teresa R.; Polvi, Elizabeth J.; Robbins, Nicole

    2017-01-01

    ABSTRACT Protein kinases are key regulators of signal transduction pathways that participate in diverse cellular processes. In fungal pathogens, kinases regulate signaling pathways that govern drug resistance, stress adaptation, and pathogenesis. The impact of kinases on the fungal regulatory circuitry has recently garnered considerable attention in the opportunistic fungal pathogen Candida albicans, which is a leading cause of human morbidity and mortality. Complex regulatory circuitry governs the C. albicans morphogenetic transition between yeast and filamentous growth, which is a key virulence trait. Here, we report that staurosporine, a promiscuous kinase inhibitor that abrogates fungal drug resistance, also influences C. albicans morphogenesis by inducing filamentation in the absence of any other inducing cue. We further establish that staurosporine exerts its effect via the adenylyl cyclase Cyr1 and the cyclic AMP (cAMP)-dependent protein kinase A (PKA). Strikingly, filamentation induced by staurosporine does not require the known upstream regulators of Cyr1, Ras1 or Pkc1, or effectors downstream of PKA, including Efg1. We further demonstrate that Cyr1 is capable of activating PKA to enable filamentation in response to staurosporine through a mechanism that does not require degradation of the transcriptional repressor Nrg1. We establish that staurosporine-induced filamentation is accompanied by a defect in septin ring formation, implicating cell cycle kinases as potential staurosporine targets underpinning this cellular response. Thus, we establish staurosporine as a chemical probe to elucidate the architecture of cellular signaling governing fungal morphogenesis and highlight the existence of novel circuitry through which the Cyr1 and PKA govern a key virulence trait. IMPORTANCE The impact of fungal pathogens on human health is devastating. One of the most pervasive fungal pathogens is Candida albicans, which kills ~40% of people suffering from bloodstream

  17. Transcriptional programs regulated by both LEAFY and APETALA1 at the time of flower formation.

    PubMed

    Winter, Cara M; Yamaguchi, Nobutoshi; Wu, Miin-Feng; Wagner, Doris

    2015-09-01

    Two key regulators of the switch to flower formation and of flower patterning in Arabidopsis are the plant-specific helix-turn-helix transcription factor LEAFY (LFY) and the MADS box transcription factor APETALA1 (AP1). The interactions between these two transcriptional regulators are complex. AP1 is both a direct target of LFY and can act in parallel with LFY. Available genetic and molecular evidence suggests that LFY and AP1 together orchestrate the switch to flower formation and early events during flower morphogenesis by altering transcriptional programs. However, very little is known about target genes regulated by both transcription factors. Here, we performed a meta-analysis of public datasets to identify genes that are likely to be regulated by both LFY and AP1. Our analyses uncovered known and novel direct LFY and AP1 targets with a role in the control of onset of flower formation. It also identified additional families of proteins and regulatory pathways that may be under transcriptional control by both transcription factors. In particular, several of these genes are linked to response to hormones, to transport and to development. Finally, we show that the gibberellin catabolism enzyme ELA1, which was recently shown to be important for the timing of the switch to flower formation, is positively feedback-regulated by AP1. Our study contributes to the elucidation of the regulatory network that leads to formation of a vital plant organ system, the flower.

  18. An invertebrate smooth muscle with striated muscle myosin filaments

    PubMed Central

    Sulbarán, Guidenn; Alamo, Lorenzo; Pinto, Antonio; Márquez, Gustavo; Méndez, Franklin; Padrón, Raúl; Craig, Roger

    2015-01-01

    Muscle tissues are classically divided into two major types, depending on the presence or absence of striations. In striated muscles, the actin filaments are anchored at Z-lines and the myosin and actin filaments are in register, whereas in smooth muscles, the actin filaments are attached to dense bodies and the myosin and actin filaments are out of register. The structure of the filaments in smooth muscles is also different from that in striated muscles. Here we have studied the structure of myosin filaments from the smooth muscles of the human parasite Schistosoma mansoni. We find, surprisingly, that they are indistinguishable from those in an arthropod striated muscle. This structural similarity is supported by sequence comparison between the schistosome myosin II heavy chain and known striated muscle myosins. In contrast, the actin filaments of schistosomes are similar to those of smooth muscles, lacking troponin-dependent regulation. We conclude that schistosome muscles are hybrids, containing striated muscle-like myosin filaments and smooth muscle-like actin filaments in a smooth muscle architecture. This surprising finding has broad significance for understanding how muscles are built and how they evolved, and challenges the paradigm that smooth and striated muscles always have distinctly different components. PMID:26443857

  19. An invertebrate smooth muscle with striated muscle myosin filaments.

    PubMed

    Sulbarán, Guidenn; Alamo, Lorenzo; Pinto, Antonio; Márquez, Gustavo; Méndez, Franklin; Padrón, Raúl; Craig, Roger

    2015-10-20

    Muscle tissues are classically divided into two major types, depending on the presence or absence of striations. In striated muscles, the actin filaments are anchored at Z-lines and the myosin and actin filaments are in register, whereas in smooth muscles, the actin filaments are attached to dense bodies and the myosin and actin filaments are out of register. The structure of the filaments in smooth muscles is also different from that in striated muscles. Here we have studied the structure of myosin filaments from the smooth muscles of the human parasite Schistosoma mansoni. We find, surprisingly, that they are indistinguishable from those in an arthropod striated muscle. This structural similarity is supported by sequence comparison between the schistosome myosin II heavy chain and known striated muscle myosins. In contrast, the actin filaments of schistosomes are similar to those of smooth muscles, lacking troponin-dependent regulation. We conclude that schistosome muscles are hybrids, containing striated muscle-like myosin filaments and smooth muscle-like actin filaments in a smooth muscle architecture. This surprising finding has broad significance for understanding how muscles are built and how they evolved, and challenges the paradigm that smooth and striated muscles always have distinctly different components.

  20. Regulation of formation of volatile compounds of tea (Camellia sinensis) leaves by single light wavelength.

    PubMed

    Fu, Xiumin; Chen, Yiyong; Mei, Xin; Katsuno, Tsuyoshi; Kobayashi, Eiji; Dong, Fang; Watanabe, Naoharu; Yang, Ziyin

    2015-11-16

    Regulation of plant growth and development by light wavelength has been extensively studied. Less attention has been paid to effect of light wavelength on formation of plant metabolites. The objective of this study was to investigate whether formation of volatiles in preharvest and postharvest tea (Camellia sinensis) leaves can be regulated by light wavelength. In the present study, in contrast to the natural light or dark treatment, blue light (470 nm) and red light (660 nm) significantly increased most endogenous volatiles including volatile fatty acid derivatives (VFADs), volatile phenylpropanoids/benzenoids (VPBs), and volatile terpenes (VTs) in the preharvest tea leaves. Furthermore, blue and red lights significantly up-regulated the expression levels of 9/13-lipoxygenases involved in VFADs formation, phenylalanine ammonialyase involved in VPBs formation, and terpene synthases involved in VTs formation. Single light wavelength had less remarkable influences on formation of volatiles in the postharvest leaves compared with the preharvest leaves. These results suggest that blue and red lights can be promising technology for remodeling the aroma of preharvest tea leaves. Furthermore, our study provided evidence that light wavelength can activate the expression of key genes involved in formation of plant volatiles for the first time.

  1. Regulation of formation of volatile compounds of tea (Camellia sinensis) leaves by single light wavelength

    PubMed Central

    Fu, Xiumin; Chen, Yiyong; Mei, Xin; Katsuno, Tsuyoshi; Kobayashi, Eiji; Dong, Fang; Watanabe, Naoharu; Yang, Ziyin

    2015-01-01

    Regulation of plant growth and development by light wavelength has been extensively studied. Less attention has been paid to effect of light wavelength on formation of plant metabolites. The objective of this study was to investigate whether formation of volatiles in preharvest and postharvest tea (Camellia sinensis) leaves can be regulated by light wavelength. In the present study, in contrast to the natural light or dark treatment, blue light (470 nm) and red light (660 nm) significantly increased most endogenous volatiles including volatile fatty acid derivatives (VFADs), volatile phenylpropanoids/benzenoids (VPBs), and volatile terpenes (VTs) in the preharvest tea leaves. Furthermore, blue and red lights significantly up-regulated the expression levels of 9/13-lipoxygenases involved in VFADs formation, phenylalanine ammonialyase involved in VPBs formation, and terpene synthases involved in VTs formation. Single light wavelength had less remarkable influences on formation of volatiles in the postharvest leaves compared with the preharvest leaves. These results suggest that blue and red lights can be promising technology for remodeling the aroma of preharvest tea leaves. Furthermore, our study provided evidence that light wavelength can activate the expression of key genes involved in formation of plant volatiles for the first time. PMID:26567525

  2. In situ transmission electron microscopy analysis of conductive filament during solid electrolyte resistance switching

    SciTech Connect

    Fujii, Takashi; Arita, Masashi; Takahashi, Yasuo; Fujiwara, Ichiro

    2011-05-23

    An in situ transmission electron microscopy (TEM) analysis of a solid electrolyte, Cu-GeS, during resistance switching is reported. Real-time observations of the filament formation and disappearance process were performed in the TEM instrument and the conductive-filament-formation model was confirmed experimentally. Narrow conductive filaments were formed corresponding to resistance switching from high- to low-resistance states. When the resistance changed to high-resistance state, the filament disappeared. It was also confirmed by use of selected area diffractometry and energy-dispersive x-ray spectroscopy that the conductive filament was made of nanocrystals composed mainly of Cu.

  3. Pneumococcal galactose catabolism is controlled by multiple regulators acting on pyruvate formate lyase

    PubMed Central

    Al-Bayati, Firas A. Y.; Kahya, Hasan F. H.; Damianou, Andreas; Shafeeq, Sulman; Kuipers, Oscar P.; Andrew, Peter W.; Yesilkaya, Hasan

    2017-01-01

    Catabolism of galactose by Streptococcus pneumoniae alters the microbe’s metabolism from homolactic to mixed acid fermentation, and this shift is linked to the microbe’s virulence. However, the genetic basis of this switch is unknown. Pyruvate formate lyase (PFL) is a crucial enzyme for mixed acid fermentation. Functional PFL requires the activities of two enzymes: pyruvate formate lyase activating enzyme (coded by pflA) and pyruvate formate lyase (coded by pflB). To understand the genetic basis of mixed acid fermentation, transcriptional regulation of pflA and pflB was studied. By microarray analysis of ΔpflB, differential regulation of several transcriptional regulators were identified, and CcpA, and GlnR’s role in active PFL synthesis was studied in detail as these regulators directly interact with the putative promoters of both pflA and pflB, their mutation attenuated pneumococcal growth, and their expression was induced on host-derived sugars, indicating that these regulators have a role in sugar metabolism, and multiple regulators are involved in active PFL synthesis. We also found that the influence of each regulator on pflA and pflB expression was distinct in terms of activation and repression, and environmental condition. These results show that active PFL synthesis is finely tuned, and feed-back inhibition and activation are involved. PMID:28240278

  4. Large-conductance channel formation mediated by P2X7 receptor activation is regulated through distinct intracellular signaling pathways in peritoneal macrophages and 2BH4 cells.

    PubMed

    Faria, R X; Cascabulho, C M; Reis, R A M; Alves, Luiz Anastácio

    2010-07-01

    The P2X(7) receptor (P2X7R) is a ligand-gated ATP receptor that acts as a low- and large-conductance channel (pore) and is known to be coupled to several downstream effectors. Recently, we demonstrated that the formation of a large-conductance channel associated with the P2X(7) receptor is induced by increasing the intracellular Ca(2+) concentration (Faria et al., Am J Physiol Cell Physiol 297:C28-C42, 2005). Here, we investigated the intracellular signaling pathways associated with P2X(7) large-conductance channel formation using the patch clamp technique in conjunction with fluorescent imaging and flow cytometry assays in 2BH4 cells and peritoneal macrophages. Different antagonists were applied to investigate the following pathways: Ca(2+)-calmodulin, phospholipase A, phospholipase D, phospholipase C, protein kinase C (PKC), mitogen-activated protein kinase (MAPK), MAPK/extracellular signal-regulated kinase, phosphoinositide 3-kinase (PI3K), and cytoskeletal proteins. Macroscopic ionic currents induced by 1 mM ATP were reduced by 85% in the presence of PKC antagonists. The addition of antagonists for MAPK, PI3K, and the cytoskeleton (actin, intermediary filament, and microtubule) blocked 92%, 83%, and 95% of the ionic currents induced by 1 mM ATP, respectively. Our results show that PKC, MAPK, PI3K, and cytoskeletal components are involved in P2X(7) receptor large-channel formation in 2BH4 cells and peritoneal macrophages.

  5. Dissection of Filamentous Growth by Transposon Mutagenesis in Saccharomyces Cerevisiae

    PubMed Central

    Mosch, H. U.; Fink, G. R.

    1997-01-01

    Diploid Saccharomyces cerevisiae strains starved for nitrogen undergo a developmental transition from growth as single yeast form (YF) cells to a multicellular form consisting of filaments of pseudohyphal (PH) cells. Filamentous growth is regulated by an evolutionarily conserved signaling pathway that includes the small GTP-binding proteins Ras2p and Cdc42p, the protein kinases Ste20p, Ste11p and Ste7p, and the transcription factor Ste12p. Here, we designed a genetic screen for mutant strains defective for filamentous growth (dfg) to identify novel targets of the filamentation signaling pathway, and we thereby identified 16 different genes, CDC39, STE12, TEC1, WHI3, NAB1, DBR1, CDC55, SRV2, TPM1, SPA2, BNI1, DFG5, DFG9, DFG10, BUD8 and DFG16, mutations that block filamentous growth. Phenotypic analysis of dfg mutant strains genetically dissects filamentous growth into the cellular processes of signal transduction, bud site selection, cell morphogenesis and invasive growth. Epistasis tests between dfg mutant alleles and dominant activated alleles of the RAS2 and STE11 genes, RAS2(Val19) and STE11-4, respectively, identify putative targets for the filamentation signaling pathway. Several of the genes described here have homologues in filamentous fungi, where they also regulate fungal development. PMID:9055077

  6. Deletions in epidermal keratins leading to alterations in filament organization in vivo and in intermediate filament assembly in vitro

    PubMed Central

    1990-01-01

    To investigate the sequences important for assembly of keratins into 10- nm filaments, we used a combined approach of (a) transfection of mutant keratin cDNAs into epithelial cells in vivo, and (b) in vitro assembly of mutant and wild-type keratins. Keratin K14 mutants missing the nonhelical carboxy- and amino-terminal domains not only integrated without perturbation into endogenous keratin filament networks in vivo, but they also formed 10-nm filaments with K5 in vitro. Surprisingly, keratin mutants missing the highly conserved L L E G E sequence, common to all intermediate filament proteins and found at the carboxy end of the alpha-helical rod domain, also assembled into filaments with only a somewhat reduced efficiency. Even a carboxy K14 mutant missing approximately 10% of the rod assembled into filaments, although in this case filaments aggregated significantly. Despite the ability of these mutants to form filaments in vitro, they often perturbed keratin filament organization in vivo. In contrast, small truncations in the amino-terminal end of the rod domain more severely disrupted the filament assembly process in vitro as well as in vivo, and in particular restricted elongation. For both carboxy and amino rod deletions, the more extensive the deletion, the more severe the phenotype. Surprisingly, while elongation could be almost quantitatively blocked with large mutations, tetramer formation and higher ordered lateral interactions still occurred. Collectively, our in vitro data (a) provide a molecular basis for the dominance of our mutants in vivo, (b) offer new insights as to why different mutants may generate different phenotypes in vivo, and (c) delineate the limit sequences necessary for K14 to both incorporate properly into a preexisting keratin filament network in vivo and assemble efficiently into 10-nm keratin filaments in vitro. PMID:1702787

  7. Somite-Derived Retinoic Acid Regulates Zebrafish Hematopoietic Stem Cell Formation

    PubMed Central

    Pillay, Laura M.; Mackowetzky, Kacey J.; Widen, Sonya A.; Waskiewicz, Andrew Jan

    2016-01-01

    Hematopoietic stem cells (HSCs) are multipotent progenitors that generate all vertebrate adult blood lineages. Recent analyses have highlighted the importance of somite-derived signaling factors in regulating HSC specification and emergence from dorsal aorta hemogenic endothelium. However, these factors remain largely uncharacterized. We provide evidence that the vitamin A derivative retinoic acid (RA) functions as an essential regulator of zebrafish HSC formation. Temporal analyses indicate that RA is required for HSC gene expression prior to dorsal aorta formation, at a time when the predominant RA synthesis enzyme, aldh1a2, is strongly expressed within the paraxial mesoderm and somites. Previous research implicated the Cxcl12 chemokine and Notch signaling pathways in HSC formation. Consequently, to understand how RA regulates HSC gene expression, we surveyed the expression of components of these pathways in RA-depleted zebrafish embryos. During somitogenesis, RA-depleted embryos exhibit altered expression of jam1a and jam2a, which potentiate Notch signaling within nascent endothelial cells. RA-depleted embryos also exhibit a severe reduction in the expression of cxcr4a, the predominant Cxcl12b receptor. Furthermore, pharmacological inhibitors of RA synthesis and Cxcr4 signaling act in concert to reduce HSC formation. Our analyses demonstrate that somite-derived RA functions to regulate components of the Notch and Cxcl12 chemokine signaling pathways during HSC formation. PMID:27861498

  8. PPM1D controls nucleolar formation by up-regulating phosphorylation of nucleophosmin

    PubMed Central

    Kozakai, Yuuki; Kamada, Rui; Furuta, Junya; Kiyota, Yuhei; Chuman, Yoshiro; Sakaguchi, Kazuyasu

    2016-01-01

    An increase of nucleolar number and size has made nucleoli essential markers for cytology and tumour development. However, the underlying basis for their structural integrity and abundance remains unclear. Protein phosphatase PPM1D was found to be up-regulated in different carcinomas including breast cancers. Here, we demonstrate for the first time that PPM1D regulates nucleolar formation via inducing an increased phosphorylation of the nucleolar protein NPM. We show that PPM1D overexpression induces an increase in the nucleolar number regardless of p53 status. We also demonstrated that specific sequential phosphorylation of NPM is important for nucleolar formation and that PPM1D is a novel upstream regulator of this phosphorylation pathway. These results enhance our understanding of the molecular mechanisms that govern nucleoli formation by demonstrating that PPM1D regulates nucleolar formation by regulating NPM phosphorylation status through a novel signalling pathway, PPM1D-CDC25C-CDK1-PLK1. PMID:27619510

  9. Osterix regulates tooth root formation in a site-specific manner.

    PubMed

    Kim, T H; Bae, C H; Lee, J C; Kim, J E; Yang, X; de Crombrugghe, B; Cho, E S

    2015-03-01

    Bone and dentin share similar biochemical compositions and physiological properties. Dentin, a major tooth component, is formed by odontoblasts; in contrast, bone is produced by osteoblasts. Osterix (Osx), a zinc finger-containing transcription factor, has been identified as an essential regulator of osteoblast differentiation and bone formation. However, it has been difficult to establish whether Osx functions in odontoblast differentiation and dentin formation. To understand the role of Osx in dentin formation, we analyzed mice in which Osx was subjected to tissue-specific ablation under the control of either the Col1a1 or the OC promoter. Two independent Osx conditional knockout mice exhibited similar molar abnormalities. Although no phenotype was found in the crowns of these teeth, both mutant lines exhibited short molar roots due to impaired root elongation. Furthermore, the interradicular dentin in these mice showed severe hypoplastic features, which were likely caused by disruptions in odontoblast differentiation and dentin formation. These phenotypes were closely related to the temporospatial expression pattern of Osx during tooth development. These findings indicate that Osx is required for root formation by regulating odontoblast differentiation, maturation, and root elongation. Cumulatively, our data strongly indicate that Osx is a site-specific regulator in tooth root formation.

  10. Particles trajectories in magnetic filaments

    SciTech Connect

    Bret, A.

    2015-07-15

    The motion of a particle in a spatially harmonic magnetic field is a basic problem involved, for example, in the mechanism of formation of a collisionless shock. In such settings, it is generally reasoned that particles entering a Weibel generated turbulence are trapped inside it, provided their Larmor radius in the peak field is smaller than the field coherence length. The goal of this work is to put this heuristic conclusion on firm ground by studying, both analytically and numerically, such motion. A toy model is analyzed, consisting of a relativistic particle entering a region of space occupied by a spatially harmonic field. The particle penetrates the magnetic structure in a direction aligned with the magnetic filaments. Although the conclusions are not trivial, the main result is confirmed.

  11. Coexistence of two types of metal filaments in oxide memristors

    NASA Astrophysics Data System (ADS)

    Xu, D.; Shangguan, X. N.; Wang, S. M.; Cao, H. T.; Liang, L. Y.; Zhang, H. L.; Gao, J. H.; Long, W. M.; Wang, J. R.; Zhuge, F.

    2017-02-01

    One generally considers the conducting filament in ZnO-based valence change memristors (VCMs) as an aggregation of oxygen vacancies. Recently, the transmission electron microscopy observation showed the filament is composed of a Zn-dominated ZnOx. In this study, careful analysis of the temperature dependence of the ON state resistance demonstrates that the formation/rupture of a Zn filament is responsible for the resistive switching in ZnO VCMs. Cu/ZnO/Pt memristive devices can be operated in both VCM and ECM (electrochemical metallization memristor) modes by forming different metal filaments including Cu, Zn and a coexistence of these two filaments. The device operation can be reversibly switched between ECM and VCM modes. The dual mode operation capability of Cu/ZnO/Pt provides a wide choice of select devices for constructing memristive crossbar architectures.

  12. Electrical contact resistance in filaments

    NASA Astrophysics Data System (ADS)

    Wu, Xiang-Fa; Zhou, Zhengping; Zhou, Wang-Min

    2012-05-01

    Electrical contact resistance (ECR) influences the electrochemical performance of porous electrodes made of stacked discrete materials (e.g., carbon nanotubes, nanofibers, etc.) for use in supercapacitors and rechargeable batteries. This study establishes a simple elasticity-conductivity model for the ECR of filaments in adhesive contact. The elastic deformation and size of electrical contact zone of the filaments are determined by using an adhesive contact model of filaments, and the ECR of adhesive filaments is obtained in explicit form. Dependencies of the ECR upon the filament geometries, surface energy, and elasticity are examined.

  13. INTERMEDIATE FILAMENTS IN SMOOTH MUSCLE

    PubMed Central

    Tang, Dale D.

    2008-01-01

    The intermediate filament (IF) network is one of the three cytoskeletal systems in smooth muscle. The type III IF proteins vimentin and desmin are major constituents of the network in smooth muscle cells and tissues. Lack of vimentin or desmin impairs contractile ability of various smooth muscle preparations, implying their important role for smooth muscle force development. The IF framework has long been viewed as a fixed cytostructure that solely provides mechanical integrity for the cell. However, recent studies suggest that the IF cytoskeleton is dynamic in mammalian cells in response to various external stimulation. In this review, the structure and biological properties of IF proteins in smooth muscle are summarized. The role of IF proteins in the modulation of smooth muscle force development and redistribution/translocation of signaling partners (such as p130 Crk-associated substrate, CAS) is depicted. This review also summarizes our latest understanding on how the IF network may be regulated in smooth muscle. PMID:18256275

  14. Regulation of type 1 fimbriae synthesis and biofilm formation by the transcriptional regulator LrhA of Escherichia coli.

    PubMed

    Blumer, Caroline; Kleefeld, Alexandra; Lehnen, Daniela; Heintz, Margit; Dobrindt, Ulrich; Nagy, Gábor; Michaelis, Kai; Emödy, Levente; Polen, Tino; Rachel, Reinhard; Wendisch, Volker F; Unden, Gottfried

    2005-10-01

    Type 1 fimbriae of Escherichia coli facilitate attachment to the host mucosa and promote biofilm formation on abiotic surfaces. The transcriptional regulator LrhA, which is known as a repressor of flagellar, motility and chemotaxis genes, regulates biofilm formation and expression of type 1 fimbriae. Whole-genome expression profiling revealed that inactivation of lrhA results in an increased expression of structural components of type 1 fimbriae. In vitro, LrhA bound to the promoter regions of the two fim recombinases (FimB and FimE) that catalyse the inversion of the fimA promoter, and to the invertible element itself. Translational lacZ fusions with these genes and quantification of fimE transcript levels by real-time PCR showed that LrhA influences type 1 fimbrial phase variation, primarily via activation of FimE, which is required for the ON-to-OFF transition of the fim switch. Enhanced type 1 fimbrial expression as a result of lrhA disruption was confirmed by mannose-sensitive agglutination of yeast cells. Biofilm formation was stimulated by lrhA inactivation and completely suppressed upon LrhA overproduction. The effects of LrhA on biofilm formation were exerted via the changed levels of surface molecules, most probably both flagella and type 1 fimbriae. Together, the data show a role for LrhA as a repressor of type 1 fimbrial expression, and thus as a regulator of the initial stages of biofilm development and, presumably, bacterial adherence to epithelial host cells also.

  15. The global regulator GacS regulates biofilm formation in Pseudomonas chlororaphis O6 differently with carbon source.

    PubMed

    Kim, Ji Soo; Kim, Yong Hwan; Park, Ju Yeon; Anderson, Anne J; Kim, Young Cheol

    2014-03-01

    An aggressive root colonizer, Pseudomonas chlororaphis O6 produces various secondary metabolites that impact plant health. The sensor kinase GacS is a key regulator of the expression of biocontrol-related traits. Biofilm formation is one such trait because of its role in root surface colonization. This paper focuses on the effects of carbon source on biofilm formation. In comparison with the wild type, a gacS mutant formed biofilms at a reduced level with sucrose as the major carbon source but at much higher level with mannitol in the defined medium. Biofilm formation by the gacS mutant occurred without phenazine production and in the absence of normal levels of acyl homoserine lactones, which promote biofilms with other pseudomonads. Colonization of tomato roots was similar for the wild type and gacS mutant, showing that any differences in biofilm formation in the rhizosphere were not of consequence under the tested conditions. The reduced ability of the gacS mutant to induce systemic resistance against tomato leaf mold and tomato gray mold was consistent with a lack of production of effectors, such as phenazines. These results demonstrated plasticity in biofilm formation and root colonization in the rhizosphere by a beneficial pseudomonad.

  16. Solid friction between soft filaments.

    PubMed

    Ward, Andrew; Hilitski, Feodor; Schwenger, Walter; Welch, David; Lau, A W C; Vitelli, Vincenzo; Mahadevan, L; Dogic, Zvonimir

    2015-06-01

    Any macroscopic deformation of a filamentous bundle is necessarily accompanied by local sliding and/or stretching of the constituent filaments. Yet the nature of the sliding friction between two aligned filaments interacting through multiple contacts remains largely unexplored. Here, by directly measuring the sliding forces between two bundled F-actin filaments, we show that these frictional forces are unexpectedly large, scale logarithmically with sliding velocity as in solid-like friction, and exhibit complex dependence on the filaments' overlap length. We also show that a reduction of the frictional force by orders of magnitude, associated with a transition from solid-like friction to Stokes's drag, can be induced by coating F-actin with polymeric brushes. Furthermore, we observe similar transitions in filamentous microtubules and bacterial flagella. Our findings demonstrate how altering a filament's elasticity, structure and interactions can be used to engineer interfilament friction and thus tune the properties of fibrous composite materials.

  17. Setting Formative Assessments in Real-World Contexts to Facilitate Self-Regulated Learning

    ERIC Educational Resources Information Center

    Tay, Hui Yong

    2015-01-01

    Some writers (Black and Wiliam in "Phi Delta Kappan" 80(2):139-148, 1998; Clark 2012; Panadero and Jonsson in "Educational Research Review" 9:129-144, 2013) have hypothesized a link between formative assessments (FA) and self-regulated learning (SRL). FA give students an opportunity to play an active role in their learning…

  18. VE-cadherin interacts with cell polarity protein Pals1 to regulate vascular lumen formation.

    PubMed

    Brinkmann, Benjamin F; Steinbacher, Tim; Hartmann, Christian; Kummer, Daniel; Pajonczyk, Denise; Mirzapourshafiyi, Fatemeh; Nakayama, Masanori; Weide, Thomas; Gerke, Volker; Ebnet, Klaus

    2016-09-15

    Blood vessel tubulogenesis requires the formation of stable cell-to-cell contacts and the establishment of apicobasal polarity of vascular endothelial cells. Cell polarity is regulated by highly conserved cell polarity protein complexes such as the Par3-aPKC-Par6 complex and the CRB3-Pals1-PATJ complex, which are expressed by many different cell types and regulate various aspects of cell polarity. Here we describe a functional interaction of VE-cadherin with the cell polarity protein Pals1. Pals1 directly interacts with VE-cadherin through a membrane-proximal motif in the cytoplasmic domain of VE-cadherin. VE-cadherin clusters Pals1 at cell-cell junctions. Mutating the Pals1-binding motif in VE-cadherin abrogates the ability of VE-cadherin to regulate apicobasal polarity and vascular lumen formation. In a similar way, deletion of the Par3-binding motif at the C-terminus of VE-cadherin impairs apicobasal polarity and vascular lumen formation. Our findings indicate that the biological activity of VE-cadherin in regulating endothelial polarity and vascular lumen formation is mediated through its interaction with the two cell polarity proteins Pals1 and Par3.

  19. VE-cadherin interacts with cell polarity protein Pals1 to regulate vascular lumen formation

    PubMed Central

    Brinkmann, Benjamin F.; Steinbacher, Tim; Hartmann, Christian; Kummer, Daniel; Pajonczyk, Denise; Mirzapourshafiyi, Fatemeh; Nakayama, Masanori; Weide, Thomas; Gerke, Volker; Ebnet, Klaus

    2016-01-01

    Blood vessel tubulogenesis requires the formation of stable cell-to-cell contacts and the establishment of apicobasal polarity of vascular endothelial cells. Cell polarity is regulated by highly conserved cell polarity protein complexes such as the Par3-aPKC-Par6 complex and the CRB3-Pals1-PATJ complex, which are expressed by many different cell types and regulate various aspects of cell polarity. Here we describe a functional interaction of VE-cadherin with the cell polarity protein Pals1. Pals1 directly interacts with VE-cadherin through a membrane-proximal motif in the cytoplasmic domain of VE-cadherin. VE-cadherin clusters Pals1 at cell–cell junctions. Mutating the Pals1-binding motif in VE-cadherin abrogates the ability of VE-cadherin to regulate apicobasal polarity and vascular lumen formation. In a similar way, deletion of the Par3-binding motif at the C-terminus of VE-cadherin impairs apicobasal polarity and vascular lumen formation. Our findings indicate that the biological activity of VE-cadherin in regulating endothelial polarity and vascular lumen formation is mediated through its interaction with the two cell polarity proteins Pals1 and Par3. PMID:27466317

  20. Up-regulation of glycolytic metabolism is required for HIF1α-driven bone formation.

    PubMed

    Regan, Jenna N; Lim, Joohyun; Shi, Yu; Joeng, Kyu Sang; Arbeit, Jeffrey M; Shohet, Ralph V; Long, Fanxin

    2014-06-10

    The bone marrow environment is among the most hypoxic in the body, but how hypoxia affects bone formation is not known. Because low oxygen tension stabilizes hypoxia-inducible factor alpha (HIFα) proteins, we have investigated the effect of expressing a stabilized form of HIF1α in osteoblast precursors. Brief stabilization of HIF1α in SP7-positive cells in postnatal mice dramatically stimulated cancellous bone formation via marked expansion of the osteoblast population. Remarkably, concomitant deletion of vascular endothelial growth factor A (VEGFA) in the mouse did not diminish bone accrual caused by HIF1α stabilization. Thus, HIF1α-driven bone formation is independent of VEGFA up-regulation and increased angiogenesis. On the other hand, HIF1α stabilization stimulated glycolysis in bone through up-regulation of key glycolytic enzymes including pyruvate dehydrogenase kinase 1 (PDK1). Pharmacological inhibition of PDK1 completely reversed HIF1α-driven bone formation in vivo. Thus, HIF1α stimulates osteoblast formation through direct activation of glycolysis, and alterations in cellular metabolism may be a broadly applicable mechanism for regulating cell differentiation.

  1. Regional orientation of actin filaments in the pericanalicular cytoplasm of rat hepatocytes.

    PubMed

    Ishii, M; Washioka, H; Tonosaki, A; Toyota, T

    1991-12-01

    To elucidate how actin filaments participate in bile formation, polarity of actin filaments in the pericanalicular cytoplasm was determined with myosin subfragment 1 by transmission electron microscopy of ultrathin sections and deep-etching replicas. Densely concentrated actin filaments were identified around the bile canaliculi in the forms of microvillous core filaments, pericanalicular web filaments, and filaments on the junctional complex. They bound subfragment 1 to form double-helical strands on the deep-etching replica or typical arrowheads on the ultrathin section. All microvillous core filaments showed their arrowheads pointing basally, suggesting the molecular growth occurring at their apical ends. In contrast, filaments of the pericanalicular web, running in parallel to the cell surface, showed unfixed polarities as indicated by their arrowheads. Furthermore, neighboring filament pairs often showed opposite polarities, an alignment necessary for filament sliding. The junctional complex had filaments with arrowheads pointed mostly at the cell center with a small number in opposite direction. In addition, a group of sporadic filaments appeared to be installed to link to both the canalicular membrane and coated vesicles. Such regionally specialized actin filaments are considered inclusively to form a cytoskeletal system that is in charge of (a) maintenance of length of the microvilli, (b) contraction of the canalicular walls, and (c) translocation of coated vesicles in the pericanalicular cytoplasm.

  2. VCP and ATL1 regulate endoplasmic reticulum and protein synthesis for dendritic spine formation.

    PubMed

    Shih, Yu-Tzu; Hsueh, Yi-Ping

    2016-03-17

    Imbalanced protein homeostasis, such as excessive protein synthesis and protein aggregation, is a pathogenic hallmark of a range of neurological disorders. Here, using expression of mutant proteins, a knockdown approach and disease mutation knockin mice, we show that VCP (valosin-containing protein), together with its cofactor P47 and the endoplasmic reticulum (ER) morphology regulator ATL1 (Atlastin-1), regulates tubular ER formation and influences the efficiency of protein synthesis to control dendritic spine formation in neurons. Strengthening the significance of protein synthesis in dendritic spinogenesis, the translation blocker cyclohexamide and the mTOR inhibitor rapamycin reduce dendritic spine density, while a leucine supplement that increases protein synthesis ameliorates the dendritic spine defects caused by Vcp and Atl1 deficiencies. Because VCP and ATL1 are the causative genes of several neurodegenerative and neurodevelopmental disorders, we suggest that impaired ER formation and inefficient protein synthesis are significant in the pathogenesis of multiple neurological disorders.

  3. VCP and ATL1 regulate endoplasmic reticulum and protein synthesis for dendritic spine formation

    PubMed Central

    Shih, Yu-Tzu; Hsueh, Yi-Ping

    2016-01-01

    Imbalanced protein homeostasis, such as excessive protein synthesis and protein aggregation, is a pathogenic hallmark of a range of neurological disorders. Here, using expression of mutant proteins, a knockdown approach and disease mutation knockin mice, we show that VCP (valosin-containing protein), together with its cofactor P47 and the endoplasmic reticulum (ER) morphology regulator ATL1 (Atlastin-1), regulates tubular ER formation and influences the efficiency of protein synthesis to control dendritic spine formation in neurons. Strengthening the significance of protein synthesis in dendritic spinogenesis, the translation blocker cyclohexamide and the mTOR inhibitor rapamycin reduce dendritic spine density, while a leucine supplement that increases protein synthesis ameliorates the dendritic spine defects caused by Vcp and Atl1 deficiencies. Because VCP and ATL1 are the causative genes of several neurodegenerative and neurodevelopmental disorders, we suggest that impaired ER formation and inefficient protein synthesis are significant in the pathogenesis of multiple neurological disorders. PMID:26984393

  4. Oligodendrocytes regulate formation of nodes of Ranvier via the recognition molecule OMgp

    PubMed Central

    NIE, DU-YU; MA, QUAN-HONG; LAW, JANICE W.S.; CHIA, CHERN-PANG; DHINGRA, NARENDER K.; SHIMODA, YASUSHI; YANG, WU-LIN; GONG, NENG; CHEN, QING-WEN; XU, GANG; HU, QI-DONG; CHOW, PIERCE K.H.; NG, YEE-KONG; LING, ENG-ANG; WATANABE, KAZUTADA; XU, TIAN-LE; HABIB, AMYN A.; SCHACHNER, MELITTA; XIAO, ZHI-CHENG

    2006-01-01

    The molecular mechanisms underlying the involvement of oligodendrocytes in formation of the nodes of Ranvier (NORs) remain poorly understood. Here we show that oligodendrocyte-myelin glycoprotein (OMgp) aggregates specifically at NORs. Nodal location of OMgp does not occur along demyelinated axons of either Shiverer or proteolipid protein (PLP) transgenic mice. Over-expression of OMgp in OLN-93 cells facilitates process outgrowth. In transgenic mice in which expression of OMgp is down-regulated, myelin thickness declines, and lateral oligodendrocyte loops at the node-paranode junction are less compacted and even join together with the opposite loops, which leads to shortened nodal gaps. Notably, each of these structural abnormalities plus modest down-regulation of expression of Na+ channel α subunit result in reduced conduction velocity in the spinal cords of the mutant mice. Thus, OMgp that is derived from glia has distinct roles in regulating nodal formation and function during CNS myelination. PMID:17364021

  5. Dlg5 Regulates Dendritic Spine Formation and Synaptogenesis by Controlling Subcellular N-Cadherin Localization

    PubMed Central

    Wang, Shih-Hsiu J.; Celic, Ivana; Choi, Se-Young; Riccomagno, Martin; Wang, Qiang; Sun, Lu O.; Mitchell, Sarah P.; Vasioukhin, Valera; Huganir, Richard L.

    2014-01-01

    Most excitatory synapses in the mammalian brain are formed on dendritic spines, and spine density has a profound impact on synaptic transmission, integration, and plasticity. Membrane-associated guanylate kinase (MAGUK) proteins are intracellular scaffolding proteins with well established roles in synapse function. However, whether MAGUK proteins are required for the formation of dendritic spines in vivo is unclear. We isolated a novel disc large-5 (Dlg5) allele in mice, Dlg5LP, which harbors a missense mutation in the DLG5 SH3 domain, greatly attenuating its ability to interact with the DLG5 GUK domain. We show here that DLG5 is a MAGUK protein that regulates spine formation, synaptogenesis, and synaptic transmission in cortical neurons. DLG5 regulates synaptogenesis by enhancing the cell surface localization of N-cadherin, revealing a key molecular mechanism for regulating the subcellular localization of this cell adhesion molecule during synaptogenesis. PMID:25232112

  6. Temporal self-regulation of transposition through host-independent transposase rodlet formation

    PubMed Central

    Woodard, Lauren E.; Downes, Laura M.; Lee, Yi-Chien; Kaja, Aparna; Terefe, Eyuel S.; Wilson, Matthew H.

    2017-01-01

    Transposons are highly abundant in eukaryotic genomes, but their mobilization must be finely tuned to maintain host organism fitness and allow for transposon propagation. Forty percent of the human genome is comprised of transposable element sequences, and the most abundant cut-and-paste transposons are from the hAT superfamily. We found that the hAT transposase TcBuster from Tribolium castaneum formed filamentous structures, or rodlets, in human tissue culture cells, after gene transfer to adult mice, and ex vivo in cell-free conditions, indicating that host co-factors or cellular structures were not required for rodlet formation. Time-lapsed imaging of GFP-laced rodlets in human cells revealed that they formed quickly in a dynamic process involving fusion and fission. We delayed the availability of the transposon DNA and found that transposition declined after transposase concentrations became high enough for visible transposase rodlets to appear. In combination with earlier findings for maize Ac elements, these results give insight into transposase overproduction inhibition by demonstrating that the appearance of transposase protein structures and the end of active transposition are simultaneous, an effect with implications for genetic engineering and horizontal gene transfer. PMID:27899587

  7. Regulation of bone formation and remodeling by G-protein-coupled receptor 48.

    PubMed

    Luo, Jian; Zhou, Wei; Zhou, Xin; Li, Dali; Weng, Jinsheng; Yi, Zhengfang; Cho, Sung Gook; Li, Chenghai; Yi, Tingfang; Wu, Xiushan; Li, Xiao-Ying; de Crombrugghe, Benoit; Höök, Magnus; Liu, Mingyao

    2009-08-01

    G-protein-coupled receptor (GPCR) 48 (Gpr48; Lgr4), a newly discovered member of the glycoprotein hormone receptor subfamily of GPCRs, is an orphan GPCR of unknown function. Using a knockout mouse model, we have characterized the essential roles of Gpr48 in bone formation and remodeling. Deletion of Gpr48 in mice results in a dramatic delay in osteoblast differentiation and mineralization, but not in chondrocyte proliferation and maturation, during embryonic bone formation. Postnatal bone remodeling is also significantly affected in Gpr48(-/-) mice, including the kinetic indices of bone formation rate, bone mineral density and osteoid formation, whereas the activity and number of osteoclasts are increased as assessed by tartrate-resistant acid phosphatase staining. Examination of the molecular mechanism of Gpr48 action in bone formation revealed that Gpr48 can activate the cAMP-PKA-CREB signaling pathway to regulate the expression level of Atf4 in osteoblasts. Furthermore, we show that Gpr48 significantly downregulates the expression levels of Atf4 target genes/proteins, such as osteocalcin (Ocn; Bglap2), bone sialoprotein (Bsp; Ibsp) and collagen. Together, our data demonstrate that Gpr48 regulates bone formation and remodeling through the cAMP-PKA-Atf4 signaling pathway.

  8. Ethylene regulates lateral root formation and auxin transport in Arabidopsis thaliana

    PubMed Central

    Negi, Sangeeta; Ivanchenko, Maria G; Muday, Gloria K

    2008-01-01

    Lateral root branching is a genetically defined and environmentally regulated process. Auxin is required for lateral root formation, and mutants that are altered in auxin synthesis, transport or signaling often have lateral root defects. Crosstalk between auxin and ethylene in root elongation has been demonstrated, but interactions between these hormones in the regulation of Arabidopsis lateral root formation are not well characterized. This study utilized Arabidopsis mutants altered in ethylene signaling and synthesis to explore the role of ethylene in lateral root formation. We find that enhanced ethylene synthesis or signaling, through the eto1-1 and ctr1-1 mutations, or through the application of 1-aminocyclopropane-1-carboxylic acid (ACC), negatively impacts lateral root formation, and is reversible by treatment with the ethylene antagonist, silver nitrate. In contrast, mutations that block ethylene responses, etr1-3 and ein2-5, enhance root formation and render it insensitive to the effect of ACC, even though these mutants have reduced root elongation at high ACC doses. ACC treatments or the eto1-1 mutation significantly enhance radiolabeled indole-3-acetic acid (IAA) transport in both the acropetal and the basipetal directions. ein2-5 and etr1-3 have less acropetal IAA transport, and transport is no longer regulated by ACC. DR5-GUS reporter expression is also altered by ACC treatment, which is consistent with transport differences. The aux1-7 mutant, which has a defect in an IAA influx protein, is insensitive to the ethylene inhibition of root formation. aux1-7 also has ACC-insensitive acropetal and basipetal IAA transport, as well as altered DR5-GUS expression, which is consistent with ethylene altering AUX1-mediated IAA uptake, and thereby blocking lateral root formation. PMID:18363780

  9. Plethora of transitions during breakup of liquid filaments

    PubMed Central

    Castrejón-Pita, José Rafael; Castrejón-Pita, Alfonso Arturo; Thete, Sumeet Suresh; Sambath, Krishnaraj; Hutchings, Ian M.; Hinch, John; Lister, John R.; Basaran, Osman A.

    2015-01-01

    Thinning and breakup of liquid filaments are central to dripping of leaky faucets, inkjet drop formation, and raindrop fragmentation. As the filament radius decreases, curvature and capillary pressure, both inversely proportional to radius, increase and fluid is expelled with increasing velocity from the neck. As the neck radius vanishes, the governing equations become singular and the filament breaks. In slightly viscous liquids, thinning initially occurs in an inertial regime where inertial and capillary forces balance. By contrast, in highly viscous liquids, initial thinning occurs in a viscous regime where viscous and capillary forces balance. As the filament thins, viscous forces in the former case and inertial forces in the latter become important, and theory shows that the filament approaches breakup in the final inertial–viscous regime where all three forces balance. However, previous simulations and experiments reveal that transition from an initial to the final regime either occurs at a value of filament radius well below that predicted by theory or is not observed. Here, we perform new simulations and experiments, and show that a thinning filament unexpectedly passes through a number of intermediate transient regimes, thereby delaying onset of the inertial–viscous regime. The new findings have practical implications regarding formation of undesirable satellite droplets and also raise the question as to whether similar dynamical transitions arise in other free-surface flows such as coalescence that also exhibit singularities. PMID:25825761

  10. The transcription factor p8 regulates autophagy during diapause embryo formation in Artemia parthenogenetica.

    PubMed

    Lin, Cheng; Jia, Sheng-Nan; Yang, Fan; Jia, Wen-Huan; Yu, Xiao-Jian; Yang, Jin-Shu; Yang, Wei-Jun

    2016-07-01

    Autophagy is an essential homeostatic process by which cytoplasmic components, including macromolecules and organelles, are degraded by lysosome. Increasing evidence suggests that phosphorylated AMP-activated protein kinase (p-AMPK) and target of rapamycin (TOR) play key roles in the regulation of autophagy. However, the regulation of autophagy in quiescent cells remains unclear, despite the fact that autophagy is known to be critical for normal development, regeneration, and degenerative diseases. Here, crustacean Artemia parthenogenetica was used as a model system because they produced and released encysted embryos that enter a state of obligate dormancy in cell quiescence to withstand various environmental threats. We observed that autophagy was increased before diapause stage but dropped to extremely low level in diapause cysts in Artemia. Western blot analyses indicated that the regulation of autophagy was AMPK/TOR independent during diapause embryo formation. Importantly, the level of p8 (Ar-p8), a stress-inducible transcription cofactor, was elevated at the stage just before diapause and was absent in encysted embryos, indicating that Ar-p8 may regulate autophagy. The results of Ar-p8 knockdown revealed that Ar-p8 regulated autophagy during diapause formation in Artemia. Moreover, we observed that activating transcription factors 4 and 6 (ATF4 and ATF6) responded to Ar-p8-regulated autophagy, indicating that autophagy targeted endoplasmic reticulum (ER) during diapause formation in Artemia. Additionally, AMPK/TOR-independent autophagy was validated in human gastric cancer MKN45 cells overexpressing Ar-p8. The findings presented here may provide insights into the role of p8 in regulating autophagy in quiescent cells.

  11. Self-assembly of Artificial Actin Filaments

    NASA Astrophysics Data System (ADS)

    Grosenick, Christopher; Cheng, Shengfeng

    Actin Filaments are long, double-helical biopolymers that make up the cytoskeleton along with microtubules and intermediate filaments. In order to further understand the self-assembly process of these biopolymers, a model to recreate actin filament geometry was developed. A monomer in the shape of a bent rod with vertical and lateral binding sites was designed to assemble into single or double helices. With Molecular Dynamics simulations, a variety of phases were observed to form by varying the strength of the binding sites. Ignoring lateral binding sites, we have found a narrow range of binding strengths that lead to long single helices via various growth pathways. When lateral binding strength is introduced, double helices begin to form. These double helices self-assemble into substantially more stable structures than their single helix counterparts. We have found double helices to form long filaments at about half the vertical binding strength of single helices. Surprisingly, we have found that triple helices occasionally form, indicating the importance of structural regulation in the self-assembly of biopolymers.

  12. PERP regulates enamel formation via effects on cell–cell adhesion and gene expression

    PubMed Central

    Jheon, Andrew H.; Mostowfi, Pasha; Snead, Malcolm L.; Ihrie, Rebecca A.; Sone, Eli; Pramparo, Tiziano; Attardi, Laura D.; Klein, Ophir D.

    2011-01-01

    Little is known about the role of cell–cell adhesion in the development of mineralized tissues. Here we report that PERP, a tetraspan membrane protein essential for epithelial integrity, regulates enamel formation. PERP is necessary for proper cell attachment and gene expression during tooth development, and its expression is controlled by P63, a master regulator of stratified epithelial development. During enamel formation, PERP is localized to the interface between the enamel-producing ameloblasts and the stratum intermedium (SI), a layer of cells subjacent to the ameloblasts. Perp-null mice display dramatic enamel defects, which are caused, in part, by the detachment of ameloblasts from the SI. Microarray analysis comparing gene expression in teeth of wild-type and Perp-null mice identified several differentially expressed genes during enamel formation. Analysis of these genes in ameloblast-derived LS8 cells upon knockdown of PERP confirmed the role for PERP in the regulation of gene expression. Together, our data show that PERP is necessary for the integrity of the ameloblast–SI interface and that a lack of Perp causes downregulation of genes that are required for proper enamel formation. PMID:21285247

  13. DNA methylation and histone acetylation work in concert to regulate memory formation and synaptic plasticity.

    PubMed

    Miller, Courtney A; Campbell, Susan L; Sweatt, J David

    2008-05-01

    A clear understanding is developing concerning the importance of epigenetic-related molecular mechanisms in transcription-dependent long-term memory formation. Chromatin modification, in particular histone acetylation, is associated with transcriptional activation, and acetylation of histone 3 (H3) occurs in Area CA1 of the hippocampus following contextual fear conditioning training. Conversely, DNA methylation is associated with transcriptional repression, but is also dynamically regulated in Area CA1 following training. We recently reported that inhibition of the enzyme responsible for DNA methylation, DNA methyltransferase (DNMT), in the adult rat hippocampus blocks behavioral memory formation. Here, we report that DNMT inhibition also blocks the concomitant memory-associated H3 acetylation, without affecting phosphorylation of its upstream regulator, extracellular signal-regulated kinase (ERK). Interestingly, the DNMT inhibitor-induced deficit in memory consolidation, along with deficits in long-term potentiation, can be rescued by pharmacologically increasing levels of histone acetylation prior to DNMT inhibition. These observations suggest that DNMT activity is not only necessary for memory and plasticity, but that DNA methylation may work in concert with histone modifications to regulate plasticity and memory formation in the adult rat hippocampus.

  14. Complex Flare Dynamics Initiated by a Filament-Filament Interaction

    NASA Astrophysics Data System (ADS)

    Zhu, Chunming; Liu, Rui; Alexander, David; Sun, Xudong; McAteer, James

    2015-04-01

    We report on a filament eruption that led to a relatively rare filament-filament interaction event. The filaments were located at different heights above the same segment of a circular polarity inversion line (PIL) around a condensed leading sunspot. The onset of the eruption of the lower of the two filaments was accompanied by a simultaneous descent of the upper filament resulting in a convergence and direct interaction of the two filaments. The interaction led to the subsequent merger of the filaments into a single magnetically complex structure that erupted to create a large solar flare and an array of complex dynamical activity. A hard X-ray coronal source and an associated enhancement of hot plasma are observed at the interface between the two interacting filaments. These phenomena are related to the production of a small C flare and the subsequent development of a much stronger M flare. Magnetic loop shrinkage and descending dark voids were observed at different locations as part of the large flare energy release giving us a unique insight into these dynamic flare phenomena.

  15. The nucleoporin ALADIN regulates Aurora A localization to ensure robust mitotic spindle formation

    PubMed Central

    Carvalhal, Sara; Ribeiro, Susana Abreu; Arocena, Miguel; Kasciukovic, Taciana; Temme, Achim; Koehler, Katrin; Huebner, Angela; Griffis, Eric R.

    2015-01-01

    The formation of the mitotic spindle is a complex process that requires massive cellular reorganization. Regulation by mitotic kinases controls this entire process. One of these mitotic controllers is Aurora A kinase, which is itself highly regulated. In this study, we show that the nuclear pore protein ALADIN is a novel spatial regulator of Aurora A. Without ALADIN, Aurora A spreads from centrosomes onto spindle microtubules, which affects the distribution of a subset of microtubule regulators and slows spindle assembly and chromosome alignment. ALADIN interacts with inactive Aurora A and is recruited to the spindle pole after Aurora A inhibition. Of interest, mutations in ALADIN cause triple A syndrome. We find that some of the mitotic phenotypes that we observe after ALADIN depletion also occur in cells from triple A syndrome patients, which raises the possibility that mitotic errors may underlie part of the etiology of this syndrome. PMID:26246606

  16. Filamentation in Air with Ultrashort Mid-Infrared Pulses

    DTIC Science & Technology

    2011-05-09

    Filamentation in air with ultrashort mid-infrared pulses Bonggu Shim,1,2 Samuel E. Schrauth,1 and Alexander L. Gaeta1,3 1School of Applied and...filamentation of ultrashort laser pulses in air in the mid-infrared regime under conditions in which the group-velocity dispersion (GVD) is anomalous. When a...and propagates several times its diffraction length. Compared with temporal self-compression in gases due to plasma formation and pulse splitting in the

  17. Burkholderia cenocepacia ShvR-regulated genes that influence colony morphology, biofilm formation, and virulence.

    PubMed

    Subramoni, Sujatha; Nguyen, David T; Sokol, Pamela A

    2011-08-01

    Burkholderia cenocepacia is an opportunistic pathogen that primarily infects cystic fibrosis (CF) patients. Previously, we reported that ShvR, a LysR regulator, influences colony morphology, virulence, and biofilm formation and regulates the expression of an adjacent 24-kb genomic region encoding 24 genes. In this study, we report the functional characterization of selected genes in this region. A Tn5 mutant with shiny colony morphology was identified with a polar mutation in BCAS0208, predicted to encode an acyl-coenzyme A dehydrogenase. Mutagenesis of BCAS0208 and complementation analyses revealed that BCAS0208 is required for rough colony morphology, biofilm formation, and virulence on alfalfa seedlings. It was not possible to complement with BCAS0208 containing a mutation in the catalytic site. BCAS0201, encoding a putative flavin adenine dinucleotide (FAD)-dependent oxidoreductase, and BCAS0207, encoding a putative citrate synthase, do not influence colony morphology but are required for optimum levels of biofilm formation and virulence. Both BCAS0208 and BCAS0201 contribute to pellicle formation, although individual mutations in each of these genes had no appreciable effect on pellicle formation. A mutant with a polar insertion in BCAS0208 was significantly less virulent in a rat model of chronic lung infection as well as in the alfalfa model. Genes in this region were shown to influence utilization of branched-chain fatty acids, tricarboxylic acid cycle substrates, l-arabinose, and branched-chain amino acids. Together, our data show that the ShvR-regulated genes BCAS0208 to BCAS0201 are required for the rough colony morphotype, biofilm and pellicle formation, and virulence in B. cenocepacia.

  18. MARCKS actin-binding capacity mediates actin filament assembly during mitosis in human hepatic stellate cells.

    PubMed

    Rombouts, Krista; Mello, Tommaso; Liotta, Francesco; Galli, Andrea; Caligiuri, Alessandra; Annunziato, Francesco; Pinzani, Massimo

    2012-08-15

    Cross-linking between the actin cytoskeleton and plasma membrane actin-binding proteins is a key interaction responsible for the mechanical properties of the mitotic cell. Little is known about the identity, the localization, and the function of actin filament-binding proteins during mitosis in human hepatic stellate cells (hHSC). The aim of the present study was to identify and analyze the cross talk between actin and myristoylated alanine-rich kinase C substrate (MARCKS), an important PKC substrate and actin filament-binding protein, during mitosis in primary hHSC. Confocal analysis and chromosomal fraction analysis of mitotic hHSC demonstrated that phosphorylated (P)-MARCKS displays distinct phase-dependent localizations, accumulates at the perichromosomal layer, and is a centrosomal protein belonging to the chromosomal cytosolic fraction. Aurora B kinase (AUBK), an important mitotic regulator, β-actin, and P-MARCKS concentrate at the cytokinetic midbody during cleavage furrow formation. This localization is critical since MARCKS-depletion in hHSC is characterized by a significant loss in cytosolic actin filaments and cortical β-actin that induces cell cycle inhibition and dislocation of AUBK. A depletion of AUBK in hHSC affects cell cycle, resulting in multinucleation. Quantitative live cell imaging demonstrates that the actin filament-binding capacity of MARCKS is key to regulate mitosis since the cell cycle inhibitory effect in MARCKS-depleted cells caused abnormal cell morphology and an aberrant cytokinesis, resulting in a significant increase in cell cycle time. These findings implicate that MARCKS, an important PKC substrate, is essential for proper cytokinesis and that MARCKS and its partner actin are key mitotic regulators during cell cycle in hHSC.

  19. Filament wound structure and method

    DOEpatents

    Dritt, William S.; Gerth, Howard L.; Knight, Jr., Charles E.; Pardue, Robert M.

    1977-01-01

    The present invention relates to a filament wound spherical structure comprising a plurality of filament band sets disposed about the surface of a mandrel with each band of each set formed of a continuous filament circumferentially wound about the mandrel a selected number of circuits and with each circuit of filament being wound parallel to and contiguous with an immediate previously wound circuit. Each filament band in each band set is wound at the same helix angle from the axis of revolution of the mandrel and all of the bands of each set are uniformly distributed about the mandrel circumference. The pole-to-equator wall thickness taper associated with each band set, as several contiguous band sets are wound about the mandrel starting at the poles, is accumulative as the band sets are nested to provide a complete filament wound sphere of essentially uniform thickness.

  20. Contractile properties of thin (actin) filament-reconstituted muscle fibers.

    PubMed

    Ishiwata, S; Funatsu, T; Fujita, H

    1998-01-01

    Selective removal and reconstitution of the components of muscle fibers (fibrils) is a useful means of examining the molecular mechanism underlying the formation of the contractile apparatus. In addition, this approach is powerful for examining the structure-function relationship of a specific component of the contractile system. In previous studies, we have achieved the partial structural and functional reconstitution of thin filaments in the skeletal contractile apparatus and full reconstitution in the cardiac contractile apparatus. First, all thin filaments other than short fragments at the Z line were removed by treatment with plasma gelsolin, an actin filament-severing protein. Under these conditions, no active tension could be generated. By incorporating exogenous actin into these thin filament-free fibers, actin filaments were reconstituted by polymerization on the short actin fragments remaining at the Z line, and active tension, which was insensitive to Ca2+, was restored. The active tension after the reconstitution of thin filaments reached as high as 30% of the original level in skeletal muscle, while it reached 140% in cardiac muscle. The augmentation of tension in cardiac muscle is mainly attributable to the elongation of reconstituted filaments, longer than the average length of thin filaments in an intact muscle. These results indicate that a muscle contractile apparatus with a high order structure and function can be constructed by the self-assembly of constituent proteins. Recently, we applied this reconstitution system to the study of the mechanism of spontaneous oscillatory contraction (SPOC) in thin (actin) filament-reconstituted cardiac muscle fibers. As a result, we found that SPOC occurs even in regulatory protein-free actin filament-reconstituted fibers (Fujita & Ishiwata, manuscript submitted), although the SPOC conditions were slightly different from the standard SPOC conditions. This result strongly suggests that spontaneous oscillation

  1. A study of short wave instability on vortex filaments

    SciTech Connect

    Wang, Hong Yun

    1996-12-01

    The numerical stability and accuracy of the vortex method are studied. The effect of the ordinary differential equations (ODE) solver and of the time step on the numerical stability is analyzed. Various ODE solvers are compared and a best performer is chosen. A new constraint on the time step based on numerical stability is proposed and verified in numerical simulations. It is shown through numerical examples that empirical rules for selecting the spatial discretization obtained in simple test problems may not be extended to more general problems. The thin tube vortex filament method is applied to the problem of Widnall's instability on vortex rings. Numerical results different from previous calculations are presented and the source of the discrepancies is explained. The long time behavior of the unstable mode on thin vortex rings is simulated and analyzed. The short wave instability on vortex filaments is investigated both theoretically and numerically. It is shown that the short wave instability always occurs on co-rotating vortex filaments of fixed core structure. Furthermore when they are close to each other, vortex filaments produce short wave unstable modes which lead to wild stretching and folding. However, when the inter-filament distance is large in comparison with the core size of the filaments, unstable modes are bounded by a small fraction of the core size and the vortex filaments do not create hairpins nor wild stretching. These findings may explain the smooth behavior of the superfluid vortices. The formation of hairpin structures on numerical vortex filaments is investigated. It is shown that the formation of hairpin structures is independent of the ODE solver, of the time step and of other numerical parameters. The hairpin structures are primarily caused by short wave instability on co-rotating vortex filaments.

  2. Large-scale filaments associated with Milky Way spiral arms

    NASA Astrophysics Data System (ADS)

    Wang, Ke; Testi, Leonardo; Ginsburg, Adam; Walmsley, C. Malcolm; Molinari, Sergio; Schisano, Eugenio

    2015-07-01

    The ubiquity of filamentary structure at various scales throughout the Galaxy has triggered a renewed interest in their formation, evolution, and role in star formation. The largest filaments can reach up to Galactic scale as part of the spiral arm structure. However, such large-scale filaments are hard to identify systematically due to limitations in identifying methodology (i.e. as extinction features). We present a new approach to directly search for the largest, coldest, and densest filaments in the Galaxy, making use of sensitive Herschel Hi-GAL (Herschel Infrared Galactic Plane Survey) data complemented by spectral line cubes. We present a sample of the nine most prominent Herschel filaments, including six identified from a pilot search field plus three from outside the field. These filaments measure 37-99 pc long and 0.6-3.0 pc wide with masses (0.5-8.3) × 104 M⊙, and beam-averaged (28 arcsec, or 0.4-0.7 pc) peak H2 column densities of (1.7-9.3)× 1022 cm- 2. The bulk of the filaments are relatively cold (17-21 K), while some local clumps have a dust temperature up to 25-47 K. All the filaments are located within ≲60 pc from the Galactic mid-plane. Comparing the filaments to a recent spiral arm model incorporating the latest parallax measurements, we find that 7/9 of them reside within arms, but most are close to arm edges. These filaments are comparable in length to the Galactic scaleheight and therefore are not simply part of a grander turbulent cascade.

  3. A Model Formative Assessment Strategy to Promote Student-Centered Self-Regulated Learning in Higher Education

    ERIC Educational Resources Information Center

    Bose, Jayakumar; Rengel, Zed

    2009-01-01

    Adult learners are already involved in the process of self-regulation; hence, higher education institutions should focus on strengthening students' self-regulatory skills. Self-regulation can be facilitated through formative assessment. This paper proposes a model formative assessment strategy that would complement existing university teaching,…

  4. Predicting Solar Filament Eruptions with HEK Filament Metadata

    NASA Astrophysics Data System (ADS)

    Aggarwal, A.; Reeves, K.; Schanche, N.

    2015-12-01

    Solar filaments are cool, dark channels of partially-ionized plasma that lie above the chromosphere. Their structure follows the neutral line between local regions of opposite magnetic polarity. Previous research (e.g. Schmieder et al. 2013) has shown a positive correlation (80%) between the occurrence of filament eruptions and coronal mass ejections (CME's). If certain filament properties, such as length, chirality, and tilt, indicate a tendency towards filament eruptions, one may be able to further predict an oncoming CME. Towards this end, we present a novel algorithm based on spatiotemporal analysis that systematically correlates filament eruptions documented in the Heliophysics Event Knowledgebase (HEK) with HEK filaments that have been grouped together using a tracking algorithm developed at Georgia State University (e.g. Kempton et al. 2014). We also find filament tracks that are not correlated with eruptions to form a null data set in a similar fashion. Finally, we compare the metadata from erupting and non-erupting filament tracks to discover which filament properties may present signs of an eruption onset. Through statistical methods such as the two-sample Kolmogorov-Smirnov test and Random Forest Classifier, we find that a filament that is increasing in length or changing in tilt with respect to the equator may be a useful gauge to predict a filament eruption. However, the average values of length and tilt for both datasets follow similar distributions, leading us to conclude that these parameters do not indicate an eruption event. This work is supported by the NSF-REU solar physics program at SAO, grant number AGS-1263241, and NSF DIBBS grant number ACI-1443061.

  5. Current filamentation model for the Weibel/Filamentation instabilities

    NASA Astrophysics Data System (ADS)

    Ryu, Chang-Mo; Huynh, Cong Tuan; Kim, Chul Min

    2016-10-01

    A current filamentaion model for a nonrelativistic plasma with e +/e- beam has been presented together with PIC simulations, which can explain the mangetic field enhancement during the Weibel/ Filamentation instabilities. This filament model assumes the Hammer-Rostoker equilibrium. In addition, this model predicts preferential acceleration/deceleration for electron-ion plasmas depending on the injected beam to be e +/e-.

  6. The alternative splicing factor Nova2 regulates vascular development and lumen formation

    PubMed Central

    Giampietro, Costanza; Deflorian, Gianluca; Gallo, Stefania; Di Matteo, Anna; Pradella, Davide; Bonomi, Serena; Belloni, Elisa; Nyqvist, Daniel; Quaranta, Valeria; Confalonieri, Stefano; Bertalot, Giovanni; Orsenigo, Fabrizio; Pisati, Federica; Ferrero, Elisabetta; Biamonti, Giuseppe; Fredrickx, Evelien; Taveggia, Carla; Wyatt, Chris D. R.; Irimia, Manuel; Di Fiore, Pier Paolo; Blencowe, Benjamin J.; Dejana, Elisabetta; Ghigna, Claudia

    2015-01-01

    Vascular lumen formation is a fundamental step during angiogenesis; yet, the molecular mechanisms underlying this process are poorly understood. Recent studies have shown that neural and vascular systems share common anatomical, functional and molecular similarities. Here we show that the organization of endothelial lumen is controlled at the post-transcriptional level by the alternative splicing (AS) regulator Nova2, which was previously considered to be neural cell-specific. Nova2 is expressed during angiogenesis and its depletion disrupts vascular lumen formation in vivo. Similarly, Nova2 depletion in cultured endothelial cells (ECs) impairs the apical distribution and the downstream signalling of the Par polarity complex, resulting in altered EC polarity, a process required for vascular lumen formation. These defects are linked to AS changes of Nova2 target exons affecting the Par complex and its regulators. Collectively, our results reveal that Nova2 functions as an AS regulator in angiogenesis and is a novel member of the ‘angioneurins' family. PMID:26446569

  7. Cross-regulation by CrcZ RNA controls anoxic biofilm formation in Pseudomonas aeruginosa

    NASA Astrophysics Data System (ADS)

    Pusic, Petra; Tata, Muralidhar; Wolfinger, Michael T.; Sonnleitner, Elisabeth; Häussler, Susanne; Bläsi, Udo

    2016-12-01

    Pseudomonas aeruginosa (PA) can thrive in anaerobic biofilms in the lungs of cystic fibrosis (CF) patients. Here, we show that CrcZ is the most abundant PA14 RNA bound to the global regulator Hfq in anoxic biofilms grown in cystic fibrosis sputum medium. Hfq was crucial for anoxic biofilm formation. This observation complied with an RNAseq based transcriptome analysis and follow up studies that implicated Hfq in regulation of a central step preceding denitrification. CrcZ is known to act as a decoy that sequesters Hfq during relief of carbon catabolite repression, which in turn alleviates Hfq-mediated translational repression of catabolic genes. We therefore inferred that CrcZ indirectly impacts on biofilm formation by competing for Hfq. This hypothesis was supported by the findings that over-production of CrcZ mirrored the biofilm phenotype of the hfq deletion mutant, and that deletion of the crcZ gene augmented biofilm formation. To our knowledge, this is the first example where competition for Hfq by CrcZ cross-regulates an Hfq-dependent physiological process unrelated to carbon metabolism.

  8. Cross-regulation by CrcZ RNA controls anoxic biofilm formation in Pseudomonas aeruginosa

    PubMed Central

    Pusic, Petra; Tata, Muralidhar; Wolfinger, Michael T.; Sonnleitner, Elisabeth; Häussler, Susanne; Bläsi, Udo

    2016-01-01

    Pseudomonas aeruginosa (PA) can thrive in anaerobic biofilms in the lungs of cystic fibrosis (CF) patients. Here, we show that CrcZ is the most abundant PA14 RNA bound to the global regulator Hfq in anoxic biofilms grown in cystic fibrosis sputum medium. Hfq was crucial for anoxic biofilm formation. This observation complied with an RNAseq based transcriptome analysis and follow up studies that implicated Hfq in regulation of a central step preceding denitrification. CrcZ is known to act as a decoy that sequesters Hfq during relief of carbon catabolite repression, which in turn alleviates Hfq-mediated translational repression of catabolic genes. We therefore inferred that CrcZ indirectly impacts on biofilm formation by competing for Hfq. This hypothesis was supported by the findings that over-production of CrcZ mirrored the biofilm phenotype of the hfq deletion mutant, and that deletion of the crcZ gene augmented biofilm formation. To our knowledge, this is the first example where competition for Hfq by CrcZ cross-regulates an Hfq-dependent physiological process unrelated to carbon metabolism. PMID:28000785

  9. Regulation of reactionary dentin formation by odontoblasts in response to polymicrobial invasion of dentin matrix

    PubMed Central

    Charadram, Nattida; Farahani, Ramin M; Harty, Derek; Rathsam, Catherine; Swain, Michael V; Hunter, Neil

    2011-01-01

    Odontoblast synthesis of dentin proceeds through discrete but overlapping phases characterized by formation of a patterned organic matrix followed by remodelling and active mineralization. Microbial invasion of dentin in caries triggers an adaptive response by odontoblasts, culminating in formation of a structurally altered reactionary dentin, marked by biochemical and architectonic modifications including diminished tubularity. Scanning electron microscopy of the collagen framework in reactionary dentin revealed a radically modified yet highly organized meshwork as indicated by fractal and lacunarity analyses. Immuno-gold labelling demonstrated increased density and regular spatial distribution of dentin sialoprotein (DSP) in reactionary dentin. DSP contributes putative hydroxyapatite nucleation sites on the collagen scaffold. To further dissect the formation of this altered dentin matrix, the associated enzymatic machinery was investigated. Analysis of extracted dentin matrix indicated increased activity of matrix metalloproteinase-2 (MMP-2) in the reactionary zone referenced to physiologic dentin. Likewise, gene expression analysis of micro-dissected odontoblast layer revealed up-regulation of MMP-2. Parallel up-regulation of tissue inhibitor of metalloproteinase-2 (TIMP-2) and membrane type 1- matrix metalloproteinase (MT1-MMP) was observed in response to caries. Next, modulation of odontoblastic dentinogenic enzyme repertoire was addressed. In the odontoblast layer expression of Toll-like receptors was markedly altered in response to bacterial invasion. In carious teeth TLR-2 and the gene encoding the corresponding adaptor protein MyD88 were down-regulated whereas genes encoding TLR-4 and adaptor proteins TRAM and Mal/TIRAP were up-regulated. TLR-4 signalling mediated by binding of bacterial products has been linked to up-regulation of MMP-2. Further, increased expression of genes encoding components of the TGF-β signalling pathway, namely SMAD-2 and SMAD-4

  10. Overview of the regulation of disulfide bond formation in Peptide and protein folding.

    PubMed

    Hidaka, Yuji

    2014-04-01

    Disulfide bonds play a critical role in the maintenance of the native conformation of proteins under thermodynamic control. In general, disulfide bond formation is associated with protein folding, and this restricts the formation of folding intermediates such as misbridged disulfide isomers or kinetically trapped conformations, which provide important information related to how proteins fold into their native conformation. Therefore, numerous studies have focused on the structural analysis of folding intermediates in vitro. However, isolating or trapping folding intermediates, as well as the entire proteins, including mutant proteins, is not an easy task. Several chemical methods have recently been developed for examining peptide and protein folding and for producing, e.g., intact, post-translationally modified, or kinetically trapped proteins, or proteins with misbridged disulfide bonds. This overview introduces chemical methods for regulating the formation of disulfide bonds of peptides and proteins in the context of the thermodynamic and kinetic control of peptide and protein folding.

  11. Clumpy Disks as a Testbed for Feedback-regulated Galaxy Formation

    NASA Astrophysics Data System (ADS)

    Mayer, Lucio; Tamburello, Valentina; Lupi, Alessandro; Keller, Ben; Wadsley, James; Madau, Piero

    2016-10-01

    We study the dependence of fragmentation in massive gas-rich galaxy disks at z > 1 on stellar feedback schemes and hydrodynamical solvers, employing the GASOLINE2 SPH code and the lagrangian mesh-less code GIZMO in finite mass mode. Non-cosmological galaxy disk runs with the standard delayed-cooling blastwave feedback are compared with runs adopting a new superbubble feedback, which produces winds by modeling the detailed physics of supernova-driven bubbles and leads to efficient self-regulation of star formation. We find that, with blastwave feedback, massive star-forming clumps form in comparable number and with very similar masses in GASOLINE2 and GIZMO. Typical clump masses are in the range 107-108 M ⊙, lower than in most previous works, while giant clumps with masses above 109 M ⊙ are exceedingly rare. By contrast, superbubble feedback does not produce massive star-forming bound clumps as galaxies never undergo a phase of violent disk instability. In this scheme, only sporadic, unbound star-forming overdensities lasting a few tens of Myr can arise, triggered by non-linear perturbations from massive satellite companions. We conclude that there is severe tension between explaining massive star-forming clumps observed at z > 1 primarily as the result of disk fragmentation driven by gravitational instability and the prevailing view of feedback-regulated galaxy formation. The link between disk stability and star formation efficiency should thus be regarded as a key testing ground for galaxy formation theory.

  12. Differential regulation of blood vessel formation between standard and delayed bone healing.

    PubMed

    Lienau, Jasmin; Schmidt-Bleek, Katharina; Peters, Anja; Haschke, Franek; Duda, Georg N; Perka, Carsten; Bail, Hermann J; Schütze, Norbert; Jakob, Franz; Schell, Hanna

    2009-09-01

    Blood vessel formation is a prerequisite for bone healing. In this study, we tested the hypothesis that a delay in bone healing is associated with an altered regulation of blood vessel formation. A tibial osteotomy was performed in two groups of sheep and stabilized with either a rigid external fixator leading to standard healing or with a highly rotationally unstable one leading to delayed healing. At days 4, 7, 9, 11, 14, 21, and 42 after surgery, total RNA was extracted from the callus. Gene expressions of vWF, an endothelial cell marker, and of several molecules related to blood vessel formation were studied by qPCR. Furthermore, histology was performed on fracture hematoma and callus sections. Histologically, the first blood vessels were detected at day 7 in both groups. mRNA expression levels of vWF, Ang1, Ang2, VEGF, CYR61, FGF2, MMP2, and TIMP1 were distinctly lower in the delayed compared to the standard healing group at several time points. Based on differential expression patterns, days 7 and 21 postoperatively were revealed to be essential time points for vascularization of the ovine fracture callus. This work demonstrates for the first time a differential regulation of blood vessel formation between standard and mechanically induced delayed healing in a sheep osteotomy model.

  13. Transmembrane Agrin Regulates Dendritic Filopodia and Synapse Formation in Mature Hippocampal Neuron Cultures

    PubMed Central

    McCroskery, Seumas; Bailey, Allison; Lin, Lin; Daniels, Mathew P.

    2009-01-01

    The transmembrane isoform of agrin (Tm-agrin) is the predominant form expressed in the brain but its putative roles in brain development are not well understood. Recent reports have implicated Tm-agrin in the formation and stabilization of filopodia on neurites of immature central and peripheral neurons in culture. In maturing central neurons, dendritic filopodia are believed to facilitate synapse formation. In the present study we have investigated the role of Tm-agrin in regulation of dendritic filopodia and synaptogenesis in maturing cultures of hippocampal neurons. We did this by infecting the neurons with an RNAi lentivirus to deplete endogenous agrin during the developmental period when filopodia density on the dendritic arbor was high, and synapse formation was rapid. We found that dendritic filopodia density was markedly reduced, as was synapse density along dendrites. Moreover, synapse formation was more sharply reduced on dendrites of infected neurons contacted by uninfected axons than on uninfected dendrites contacted by infected axons. The results are consistent with a physiological role for Tm-agrin in the maturation of hippocampal neurons involving positive regulation of dendritic filopodia and consequent promotion of synaptogenesis, but also suggest a role for axonal agrin in synaptogenesis. PMID:19524020

  14. On the universality of interstellar filaments: theory meets simulations and observations

    NASA Astrophysics Data System (ADS)

    Federrath, Christoph

    2016-03-01

    Filaments are ubiquitous in the Universe. Recent observations have revealed that stars and star clusters form preferentially along dense filaments. Understanding the formation and properties of filaments is therefore a crucial step in understanding star formation. Here we perform 3D high-resolution magnetohydrodynamical simulations that follow the evolution of molecular clouds and the formation of filaments and stars. We apply a filament detection algorithm and compare simulations with different combinations of physical ingredients: gravity, turbulence, magnetic fields and jet/outflow feedback. We find that gravity-only simulations produce significantly narrower filament profiles than observed, while simulations that include turbulence produce realistic filament properties. For these turbulence simulations, we find a remarkably universal filament width of 0.10 ± 0.02 pc, which is independent of the star formation history of the clouds. We derive a theoretical model that provides a physical explanation for this characteristic filament width, based on the sonic scale (λsonic) of molecular cloud turbulence. Our derivation provides λsonic as a function of the cloud diameter L, the velocity dispersion σv, the gas sound speed cs, and the ratio of thermal to magnetic pressure, plasma β. For typical cloud conditions in the Milky Way spiral arms, we find λsonic = 0.04-0.16 pc, in excellent agreement with the filament width of 0.05-0.15 pc from observations. Consistent with the theoretical model assumptions, we find that the velocity dispersion inside the filaments is subsonic and supersonic outside. We further explain the observed p = 2 scaling of the filament density profile, ρ ∝ r-p with the collision of two planar shocks forming a filament at their intersection.

  15. Phosphorylation-Dependent Regulation of G-Protein Cycle during Nodule Formation in Soybean[OPEN

    PubMed Central

    2015-01-01

    Signaling pathways mediated by heterotrimeric G-protein complexes comprising Gα, Gβ, and Gγ subunits and their regulatory RGS (Regulator of G-protein Signaling) protein are conserved in all eukaryotes. We have shown that the specific Gβ and Gγ proteins of a soybean (Glycine max) heterotrimeric G-protein complex are involved in regulation of nodulation. We now demonstrate the role of Nod factor receptor 1 (NFR1)-mediated phosphorylation in regulation of the G-protein cycle during nodulation in soybean. We also show that during nodulation, the G-protein cycle is regulated by the activity of RGS proteins. Lower or higher expression of RGS proteins results in fewer or more nodules, respectively. NFR1 interacts with RGS proteins and phosphorylates them. Analysis of phosphorylated RGS protein identifies specific amino acids that, when phosphorylated, result in significantly higher GTPase accelerating activity. These data point to phosphorylation-based regulation of G-protein signaling during nodule development. We propose that active NFR1 receptors phosphorylate and activate RGS proteins, which help maintain the Gα proteins in their inactive, trimeric conformation, resulting in successful nodule development. Alternatively, RGS proteins might also have a direct role in regulating nodulation because overexpression of their phospho-mimic version leads to partial restoration of nodule formation in nod49 mutants. PMID:26498905

  16. Notch1-Dll4 signalling and mechanical force regulate leader cell formation during collective cell migration.

    PubMed

    Riahi, Reza; Sun, Jian; Wang, Shue; Long, Min; Zhang, Donna D; Wong, Pak Kin

    2015-03-13

    At the onset of collective cell migration, a subset of cells within an initially homogenous population acquires a distinct 'leader' phenotype with characteristic morphology and motility. However, the factors driving the leader cell formation as well as the mechanisms regulating leader cell density during the migration process remain to be determined. Here we use single-cell gene expression analysis and computational modelling to show that the leader cell identity is dynamically regulated by Dll4 signalling through both Notch1 and cellular stress in a migrating epithelium. Time-lapse microscopy reveals that Dll4 is induced in leader cells after the creation of the cell-free region and leader cells are regulated via Notch1-Dll4 lateral inhibition. Furthermore, mechanical stress inhibits Dll4 expression and leader cell formation in the monolayer. Collectively, our findings suggest that a reduction of mechanical force near the boundary promotes Notch1-Dll4 signalling to dynamically regulate the density of leader cells during collective cell migration.

  17. Notch1-Dll4 signaling and mechanical force regulate leader cell formation during collective cell migration

    PubMed Central

    Riahi, Reza; Sun, Jian; Wang, Shue; Long, Min; Zhang, Donna D.; Wong, Pak Kin

    2015-01-01

    At the onset of collective cell migration, a subset of cells within an initially homogenous population acquires a distinct “leader” phenotype with characteristic morphology and motility. However, the factors driving leader cell formation as well as the mechanisms regulating leader cell density during the migration process remain to be determined. Here, we use single cell gene expression analysis and computational modeling to show that leader cell identity is dynamically regulated by Dll4 signaling through both Notch1 and cellular stress in a migrating epithelium. Time-lapse microscopy reveals that Dll4 is induced in leader cells after the creation of the cell-free region and leader cells are regulated via Notch1-Dll4 lateral inhibition. Furthermore, mechanical stress inhibits Dll4 expression and leader cell formation in the monolayer. Collectively, our findings suggest that a reduction of mechanical force near the boundary promotes Notch1-Dll4 signaling to dynamically regulate the density of leader cells during collective cell migration. PMID:25766473

  18. Cyclic-di-GMP signalling regulates motility and biofilm formation in Bordetella bronchiseptica

    PubMed Central

    Sisti, Federico; Ha, Dae-Gon; O'Toole, George A.; Hozbor, Daniela

    2013-01-01

    The signalling molecule bis-(3′–5′)-cyclic-dimeric guanosine monophosphate (c-di-GMP) is a central regulator of diverse cellular functions, including motility, biofilm formation, cell cycle progression and virulence, in bacteria. Multiple diguanylate cyclase and phosphodiesterase-domain-containing proteins (GGDEF and EAL/HD-GYP, respectively) modulate the levels of the second messenger c-di-GMP to transmit signals and obtain such specific cellular responses. In the genus Bordetella this c-di-GMP network is poorly studied. In this work, we evaluated the expression of two phenotypes in Bordetella bronchiseptica regulated by c-di-GMP, biofilm formation and motility, under the influence of ectopic expression of Pseudomonas aeruginosa proteins with EAL or GGDEF domains that regulates the c-di-GMP level. In agreement with previous reports for other bacteria, we observed that B. bronchiseptica is able to form biofilm and reduce its motility only when GGDEF domain protein is expressed. Moreover we identify a GGDEF domain protein (BB3576) with diguanylate cyclase activity that participates in motility and biofilm regulation in B. bronchiseptica. These results demonstrate for the first time, to our knowledge, the presence of c-di-GMP regulatory signalling in B. bronchiseptica. PMID:23475948

  19. Orientation of endothelial cell division is regulated by VEGF signaling during blood vessel formation

    PubMed Central

    Zeng, Gefei; Taylor, Sarah M.; McColm, Janet R.; Kappas, Nicholas C.; Kearney, Joseph B.; Williams, Lucy H.; Hartnett, Mary E.; Bautch, Victoria L.

    2007-01-01

    New blood vessel formation requires the coordination of endothelial cell division and the morphogenetic movements of vessel expansion, but it is not known how this integration occurs. Here, we show that endothelial cells regulate division orientation during the earliest stages of blood vessel formation, in response to morphogenetic cues. In embryonic stem (ES) cell–derived vessels that do not experience flow, the plane of endothelial cytokinesis was oriented perpendicular to the vessel long axis. We also demonstrated regulated cleavage orientation in vivo, in flow-exposed forming retinal vessels. Daughter nuclei moved away from the cleavage plane after division, suggesting that regulation of endothelial division orientation effectively extends vessel length in these developing vascular beds. A gain-of-function mutation in VEGF signaling increased randomization of endothelial division orientation, and this effect was rescued by a transgene, indicating that regulation of division orientation is a novel mechanism whereby VEGF signaling affects vessel morphogenesis. Thus, our findings show that endothelial cell division and morphogenesis are integrated in developing vessels by flow-independent mechanisms that involve VEGF signaling, and this cross talk is likely to be critical to proper vessel morphogenesis. PMID:17068148

  20. Lipid nanotube formation using space-regulated electric field above interdigitated electrodes.

    PubMed

    Bi, Hongmei; Fu, Dingguo; Wang, Lei; Han, Xiaojun

    2014-04-22

    Lipid nanotubes have great potential in biology and nanotechnology. Here we demonstrate a method to form lipid nanotubes using space-regulated AC electric fields above coplanar interdigitated electrodes. The AC electric field distribution can be regulated by solution height above the electrodes. The ratio of field component in x axis (Ex) to field component in z axis (Ez) increases dramatically at solution height below 50 μm; therefore, at lower solution height, the force from Ex predominantly drives lipids to form lipid nanotubes along with the electric field direction. The forces exerted on the lipid nanotube during its formation were analyzed in detail, and an equation was obtained to describe the relationship among nanotube length and field frequency, amplitude, and time. We believe that the presented approach opens a way to design and prepare nanoscale materials with unique structural and functional properties using space-regulated electric fields.

  1. The Role of Magnetic Fields in High-Mass Star-Forming Filaments

    NASA Astrophysics Data System (ADS)

    Stephens, Ian

    Filaments are ubiquitous in the star formation process. Planck has revealed that magnetic fields are perpendicular to the densest filaments, which are the birthplace of high-mass stars, suggesting that fields help funnel gas into the filaments. However, the resolved field morphologies and strengths in the dense filaments are unknown. We propose HAWC+ 53 and 214 um polarimetric observations toward two filaments, the Snake (G11.1) and G18.6, to unveil the field morphology. Such observations will probe the filament field morphology at the subarcminute scale over the largest spatial extent to date: 25 and 9 pc respectively. We expect to have over 400 independent beams worth of detections. From the field morphology, we will test the hub-filament theory and investigate how the magnetic field strength and morphology changes with evolution and size-scale.

  2. Competition of multiple filaments during the propagation of intense femtosecond laser pulses

    SciTech Connect

    Hosseini, S.A.; Luo, Q.; Ferland, B.; Liu, W.; Chin, S.L.; Kosareva, O.G.; Panov, N.A.; Kandidov, V.P.; Akoezbek, N.

    2004-09-01

    We observed a universal phenomenon of the competition among multiple filaments generated during the propagation of intense femtosecond laser pulses in air. We show that the fluorescence signal from the excitation of nitrogen molecules inside the plasma channel contains important information pertaining to the formation and interaction of multiple filaments. The detected backscattered nitrogen fluorescence from inside the filaments yielded irregular changes from shot to shot which cannot be explained by fluctuation arising from the initial laser pulse itself. Numerical simulations reveal a complex dynamics of multiple filament propagation and interaction dynamics that depends strongly on the initial perturbations of the laser beam. The irregular changes of the fluorescence signal are attributed to the interference between adjacent hot spots that evolve into filaments which give rise to new hot spots (filaments) in between, and thus give the appearance of the fusion or branching of filaments.

  3. Detection of the insulating gap and conductive filament growth direction in resistive memories.

    PubMed

    Yalon, E; Karpov, I; Karpov, V; Riess, I; Kalaev, D; Ritter, D

    2015-10-07

    Filament growth is a key aspect in the operation of bipolar resistive random access memory (RRAM) devices, yet there are conflicting reports in the literature on the direction of growth of conductive filaments in valence change RRAM devices. We report here that an insulating gap between the filament and the semiconductor electrode can be detected by the metal-insulator-semiconductor bipolar transistor structure, and thus provide information on the filament growth direction. Using this technique, we show how voltage polarity and electrode chemistry control the filament growth direction during electro-forming. The experimental results and the nature of a gap between the filament and an electrode are discussed in light of possible models of filament formation.

  4. The Histidine Kinase BinK Is a Negative Regulator of Biofilm Formation and Squid Colonization

    PubMed Central

    Brooks, John F.

    2016-01-01

    ABSTRACT Bacterial colonization of animal epithelial tissue is a dynamic process that relies on precise molecular communication. Colonization of Euprymna scolopes bobtail squid by Vibrio fischeri bacteria requires bacterial aggregation in host mucus as the symbiont transitions from a planktonic lifestyle in seawater to a biofilm-associated state in the host. We have identified a gene, binK (biofilm inhibitor kinase; VF_A0360), which encodes an orphan hybrid histidine kinase that negatively regulates the V. fischeri symbiotic biofilm (Syp) in vivo and in vitro. We identified binK mutants as exhibiting a colonization advantage in a global genetic screen, a phenotype that we confirmed in controlled competition experiments. Bacterial biofilm aggregates in the host are larger in strains lacking BinK, whereas overexpression of BinK suppresses biofilm formation and squid colonization. Signaling through BinK is required for temperature modulation of biofilm formation at 28°C. Furthermore, we present evidence that BinK acts upstream of SypG, the σ54-dependent transcriptional regulator of the syp biofilm locus. The BinK effects are dependent on intact signaling in the RscS-Syp biofilm pathway. Therefore, we propose that BinK antagonizes the signal from RscS and serves as an integral component in V. fischeri biofilm regulation. IMPORTANCE Bacterial lifestyle transitions underlie the colonization of animal hosts from environmental reservoirs. Formation of matrix-enclosed, surface-associated aggregates (biofilms) is common in beneficial and pathogenic associations, but investigating the genetic basis of biofilm development in live animal hosts remains a significant challenge. Using the bobtail squid light organ as a model, we analyzed putative colonization factors and identified a histidine kinase that negatively regulates biofilm formation at the host interface. This work reveals a novel in vivo biofilm regulator that influences the transition of bacteria from their

  5. Surface manipulation of protein filaments

    NASA Astrophysics Data System (ADS)

    Kreplak, Laurent; Staple, Douglas; Loparic, Marko; Kreuzer, Hans-Juergen

    2009-03-01

    Within mammalian tissues, cells move by actively remodeling a dense network of collagen fibrils. In order to study this situation, we analyze the force response of two types of filamentous protein structures, desmin intermediate filaments 12 nm in diameter and collagen fibrils 80 nm in diameter. Both types of filaments were adsorbed at a solid-liquid interface and locally moved with an AFM tip at constant velocity against surface friction in the interfacial plane. In the case of collagen fibrils, that have an extensibility below 30% extension, we observed that microns long fibrils could be moved by the tip and deformed into shapes that could not be explain by the linear elastic theory for a stiff rod. In the case of desmin filaments that can be stretched up to 3.5 times there length, we observed local stretching of the filaments and discreet steps in the torsional force measured with the cantilever. In order to describe both types of filaments' behaviors, we described the protein filaments as a chain of beads of mass m linked together by a mass-less polymer linker. By solving the Newtonian equations of motions for the coupled beads in the presence of a point load and a viscous drag due to the surface-filament interactions we were able to reproduced our experimental data and extract information on friction.

  6. Filamentation in tokamaks

    SciTech Connect

    Cardozo, N.J.; Barth, C.J.; Chu, C.C.; Lok, J.; Montvai, A.; Oomens, A.A.; Peters, M.; Pijper, F.J.; de Rover, M.; Schueller, F.C.; Steenbakkers, M.F.; RTP team

    1995-09-01

    The relevance of a nest of toroidal flux surfaces as a paradigm of the magnetic topology of a tokamak plasma is challenged. High resolution Thomson scattering measurements of electron temperature and density in RTP show several hot filaments in the plasma center and sharp gradients near the sawtooth inversion radius and structures outside the sawtooth region under central ECH. In ohmic plasmas, too, the pressure and temperature profiles show significant bumps. These measurements give evidence of a complex magnetic topology. Transport in a medium with spatially strongly varying diffusivity is considered. It is shown that macroscopic transport is determined by the microscopic structure: a transport theory must predict this structure and the diffusivity in the insulating regions, while the {open_quote}turbulent{close_quote} diffusivity is irrelevant. A numerical approach to equilibria with broken surfaces is presented. {copyright} {ital 1995 American Institute of Physics.}

  7. Scientific considerations related to regulation development for CO2 sequestration in brine formations

    NASA Astrophysics Data System (ADS)

    Tsang, Chin-Fu; Benson, Sally M.; Kobelski, Bruce; Smith, Robert E.

    2002-06-01

    Carbon management through the underground injection of CO2 into subsurface brine formations is being actively studied. If there are no technological constraints for implementation, there could be a large number of wells constructed for injecting a large volume of CO2. It is therefore important, in parallel with current scientific studies, to consider the appropriate, science-based regulatory framework for CO2 injection. The Environmental Protection Agency (EPA) Underground Injection Control (UIC) program, authorized under the Safe Drinking Water Act (SDWA), has extensive experience in regulating the injection of mainly liquid wastes into geologic formations in the United States. The federal requirements and permit process implemented by EPA and the Primacy States since 1980 have played a critical role in the safety of subsurface disposal of liquid wastes in the US. Physically and chemically, there are significant differences between CO2 and common liquid wastes. Its viscosity and density are much lower and, under injection pressure in the deep formation, it may be under supercritical conditions. Because of the lower density and viscosity, CO2 leakage through the confining strata may be greater when compared to currently injected liquid wastes. Also, the chemical interactions of CO2 with the geologic formation have their own characteristics. All these scientific factors need to be evaluated to identify new guidelines for appropriate regulatory and monitoring controls. The paper reviews current UIC regulations, injection-well classification scheme and monitoring requirements, and identifies the unique factors related to the physical and chemical processes in the subsurface associated with CO2 injection. Implications of these scientific considerations for regulation development are discussed.

  8. Solid friction between soft filaments

    NASA Astrophysics Data System (ADS)

    Ward, Andrew; Hilitski, Feodor; Schwenger, Walter; Welch, David; Lau, A. W. C.; Vitelli, Vincenzo; Mahadevan, L.; Dogic, Zvonimir

    2015-06-01

    Any macroscopic deformation of a filamentous bundle is necessarily accompanied by local sliding and/or stretching of the constituent filaments. Yet the nature of the sliding friction between two aligned filaments interacting through multiple contacts remains largely unexplored. Here, by directly measuring the sliding forces between two bundled F-actin filaments, we show that these frictional forces are unexpectedly large, scale logarithmically with sliding velocity as in solid-like friction, and exhibit complex dependence on the filaments’ overlap length. We also show that a reduction of the frictional force by orders of magnitude, associated with a transition from solid-like friction to Stokes’s drag, can be induced by coating F-actin with polymeric brushes. Furthermore, we observe similar transitions in filamentous microtubules and bacterial flagella. Our findings demonstrate how altering a filament’s elasticity, structure and interactions can be used to engineer interfilament friction and thus tune the properties of fibrous composite materials.

  9. Solid friction between soft filaments

    SciTech Connect

    Ward, Andrew; Hilitski, Feodor; Schwenger, Walter; Welch, David; Lau, A. W. C.; Vitelli, Vincenzo; Mahadevan, L.; Dogic, Zvonimir

    2015-03-02

    Any macroscopic deformation of a filamentous bundle is necessarily accompanied by local sliding and/or stretching of the constituent filaments. Yet the nature of the sliding friction between two aligned filaments interacting through multiple contacts remains largely unexplored. Here, by directly measuring the sliding forces between two bundled F-actin filaments, we show that these frictional forces are unexpectedly large, scale logarithmically with sliding velocity as in solid-like friction, and exhibit complex dependence on the filaments’ overlap length. We also show that a reduction of the frictional force by orders of magnitude, associated with a transition from solid-like friction to Stokes’s drag, can be induced by coating F-actin with polymeric brushes. Furthermore, we observe similar transitions in filamentous microtubules and bacterial flagella. In conclusion, our findings demonstrate how altering a filament’s elasticity, structure and interactions can be used to engineer interfilament friction and thus tune the properties of fibrous composite materials.

  10. Spacecraft Formation Flying Maneuvers Using Linear-Quadratic Regulation with No Radial Axis Inputs

    NASA Technical Reports Server (NTRS)

    Starin, Scott R.; Yedavalli, R. K.; Sparks, Andrew G.; Bauer, Frank H. (Technical Monitor)

    2001-01-01

    Regarding multiple spacecraft formation flying, the observation has been made that control thrust need only be applied coplanar to the local horizon to achieve complete controllability of a two-satellite (leader-follower) formation. A formulation of orbital dynamics using the state of one satellite relative to another is used. Without the need for thrust along the radial (zenith-nadir) axis of the relative reference frame ' propulsion system simplifications and weight reduction may be accomplished. Several linear-quadratic regulators (LQR) are explored and compared based on performance measures likely to be important to many missions, but not directly optimized in the LQR designs. Maneuver simulations are performed using commercial ODE solvers to propagate the Keplerian dynamics of a controlled satellite relative to an uncontrolled leader. These short maneuver simulations demonstrate the capacity of the controller to perform changes from one formation geometry to another. This work focusses on formations in which the controlled satellite has a relative trajectory which projects onto the local horizon of the uncontrolled satellite as a circle. This formation has potential uses for distributed remote sensing systems.

  11. Inhibition of apoptosis signal-regulating kinase 1 enhances endochondral bone formation by increasing chondrocyte survival

    PubMed Central

    Eaton, G J; Zhang, Q-S; Diallo, C; Matsuzawa, A; Ichijo, H; Steinbeck, M J; Freeman, T A

    2014-01-01

    Endochondral ossification is the result of chondrocyte differentiation, hypertrophy, death and replacement by bone. The careful timing and progression of this process is important for normal skeletal bone growth and development, as well as fracture repair. Apoptosis Signal-Regulating Kinase 1 (ASK1) is a mitogen-activated protein kinase (MAPK), which is activated by reactive oxygen species and other cellular stress events. Activation of ASK1 initiates a signaling cascade known to regulate diverse cellular events including cytokine and growth factor signaling, cell cycle regulation, cellular differentiation, hypertrophy, survival and apoptosis. ASK1 is highly expressed in hypertrophic chondrocytes, but the role of ASK1 in skeletal tissues has not been investigated. Herein, we report that ASK1 knockout (KO) mice display alterations in normal growth plate morphology, which include a shorter proliferative zone and a lengthened hypertrophic zone. These changes in growth plate dynamics result in accelerated long bone mineralization and an increased formation of trabecular bone, which can be attributed to an increased resistance of terminally differentiated chondrocytes to undergo cell death. Interestingly, under normal cell culture conditions, mouse embryonic fibroblasts (MEFs) derived from ASK1 KO mice show no differences in either MAPK signaling or osteogenic or chondrogenic differentiation when compared with wild-type (WT) MEFs. However, when cultured with stress activators, H2O2 or staurosporine, the KO cells show enhanced survival, an associated decrease in the activation of proteins involved in death signaling pathways and a reduction in markers of terminal differentiation. Furthermore, in both WT mice treated with the ASK1 inhibitor, NQDI-1, and ASK1 KO mice endochondral bone formation was increased in an ectopic ossification model. These findings highlight a previously unrealized role for ASK1 in regulating endochondral bone formation. Inhibition of ASK1 has

  12. Biofilms from a Brazilian water distribution system include filamentous fungi.

    PubMed

    Siqueira, V M; Oliveira, H M B; Santos, C; Paterson, R R M; Gusmão, N B; Lima, N

    2013-03-01

    Filamentous fungi in drinking water can block water pipes, can cause organoleptic biodeterioration, and are a source of pathogens. There are increasing reports of the involvement of the organisms in biofilms. This present study describes a sampling device that can be inserted directly into pipes within water distribution systems, allowing biofilm formation in situ. Calcofluor White M2R staining and fluorescent in situ hybridization with morphological analyses using epifluorescent microscopy were used to analyse biofilms for filamentous fungi, permitting direct observation of the fungi. DAPI (4',6-diamidino-2-phenylindole) was applied to detect bacteria. Filamentous fungi were detected in biofilms after 6 months on coupons exposed to raw water, decanted water and at the entrance of the water distribution system. Algae, yeast, and bacteria were also observed. The role of filamentous fungi requires further investigations.

  13. Preserved filamentous microbial biosignatures in the Brick Flat gossan, Iron Mountain, California

    USGS Publications Warehouse

    Williams, Amy J.; Sumner, Dawn Y.; Alpers, Charles N.; Karunatillake, Suniti; Hofmann, Beda A

    2015-01-01

    A variety of actively precipitating mineral environments preserve morphological evidence of microbial biosignatures. One such environment with preserved microbial biosignatures is the oxidized portion of a massive sulfide deposit, or gossan, such as that at Iron Mountain, California. This gossan may serve as a mineralogical analogue to some ancient martian environments due to the presence of oxidized iron and sulfate species, and minerals that only form in acidic aqueous conditions, in both environments. Evaluating the potential biogenicity of cryptic textures in such martian gossans requires an understanding of how microbial textures form biosignatures on Earth. The iron-oxide-dominated composition and morphology of terrestrial, nonbranching filamentous microbial biosignatures may be distinctive of the underlying formation and preservation processes. The Iron Mountain gossan consists primarily of ferric oxide (hematite), hydrous ferric oxide (HFO, predominantly goethite), and jarosite group minerals, categorized into in situ gossan, and remobilized iron deposits. We interpret HFO filaments, found in both gossan types, as HFO-mineralized microbial filaments based in part on (1) the presence of preserved central filament lumina in smooth HFO mineral filaments that are likely molds of microbial filaments, (2) mineral filament formation in actively precipitating iron-oxide environments, (3) high degrees of mineral filament bending consistent with a flexible microbial filament template, and (4) the presence of bare microbial filaments on gossan rocks. Individual HFO filaments are below the resolution of the Mars Curiosity and Mars 2020 rover cameras, but sinuous filaments forming macroscopic matlike textures are resolvable. If present on Mars, available cameras may resolve these features identified as similar to terrestrial HFO filaments and allow subsequent evaluation for their biogenicity by synthesizing geochemical, mineralogical, and morphological analyses. Sinuous

  14. Preserved Filamentous Microbial Biosignatures in the Brick Flat Gossan, Iron Mountain, California.

    PubMed

    Williams, Amy J; Sumner, Dawn Y; Alpers, Charles N; Karunatillake, Suniti; Hofmann, Beda A

    2015-08-01

    A variety of actively precipitating mineral environments preserve morphological evidence of microbial biosignatures. One such environment with preserved microbial biosignatures is the oxidized portion of a massive sulfide deposit, or gossan, such as that at Iron Mountain, California. This gossan may serve as a mineralogical analogue to some ancient martian environments due to the presence of oxidized iron and sulfate species, and minerals that only form in acidic aqueous conditions, in both environments. Evaluating the potential biogenicity of cryptic textures in such martian gossans requires an understanding of how microbial textures form biosignatures on Earth. The iron-oxide-dominated composition and morphology of terrestrial, nonbranching filamentous microbial biosignatures may be distinctive of the underlying formation and preservation processes. The Iron Mountain gossan consists primarily of ferric oxide (hematite), hydrous ferric oxide (HFO, predominantly goethite), and jarosite group minerals, categorized into in situ gossan, and remobilized iron deposits. We interpret HFO filaments, found in both gossan types, as HFO-mineralized microbial filaments based in part on (1) the presence of preserved central filament lumina in smooth HFO mineral filaments that are likely molds of microbial filaments, (2) mineral filament formation in actively precipitating iron-oxide environments, (3) high degrees of mineral filament bending consistent with a flexible microbial filament template, and (4) the presence of bare microbial filaments on gossan rocks. Individual HFO filaments are below the resolution of the Mars Curiosity and Mars 2020 rover cameras, but sinuous filaments forming macroscopic matlike textures are resolvable. If present on Mars, available cameras may resolve these features identified as similar to terrestrial HFO filaments and allow subsequent evaluation for their biogenicity by synthesizing geochemical, mineralogical, and morphological analyses. Sinuous

  15. Preserved Filamentous Microbial Biosignatures in the Brick Flat Gossan, Iron Mountain, California

    PubMed Central

    Sumner, Dawn Y.; Alpers, Charles N.; Karunatillake, Suniti; Hofmann, Beda A.

    2015-01-01

    Abstract A variety of actively precipitating mineral environments preserve morphological evidence of microbial biosignatures. One such environment with preserved microbial biosignatures is the oxidized portion of a massive sulfide deposit, or gossan, such as that at Iron Mountain, California. This gossan may serve as a mineralogical analogue to some ancient martian environments due to the presence of oxidized iron and sulfate species, and minerals that only form in acidic aqueous conditions, in both environments. Evaluating the potential biogenicity of cryptic textures in such martian gossans requires an understanding of how microbial textures form biosignatures on Earth. The iron-oxide-dominated composition and morphology of terrestrial, nonbranching filamentous microbial biosignatures may be distinctive of the underlying formation and preservation processes. The Iron Mountain gossan consists primarily of ferric oxide (hematite), hydrous ferric oxide (HFO, predominantly goethite), and jarosite group minerals, categorized into in situ gossan, and remobilized iron deposits. We interpret HFO filaments, found in both gossan types, as HFO-mineralized microbial filaments based in part on (1) the presence of preserved central filament lumina in smooth HFO mineral filaments that are likely molds of microbial filaments, (2) mineral filament formation in actively precipitating iron-oxide environments, (3) high degrees of mineral filament bending consistent with a flexible microbial filament template, and (4) the presence of bare microbial filaments on gossan rocks. Individual HFO filaments are below the resolution of the Mars Curiosity and Mars 2020 rover cameras, but sinuous filaments forming macroscopic matlike textures are resolvable. If present on Mars, available cameras may resolve these features identified as similar to terrestrial HFO filaments and allow subsequent evaluation for their biogenicity by synthesizing geochemical, mineralogical, and morphological analyses

  16. The Pool of ADP and ATP Regulates Anaerobic Product Formation in Resting Cells of Lactococcus lactis

    PubMed Central

    Palmfeldt, Johan; Paese, Marco; Hahn-Hägerdal, Bärbel; van Niel, Ed W. J.

    2004-01-01

    Lactococcus lactis grows homofermentatively on glucose, while its growth on maltose under anaerobic conditions results in mixed acid product formation in which formate, acetate, and ethanol are formed in addition to lactate. Maltose was used as a carbon source to study mixed acid product formation as a function of the growth rate. In batch and nitrogen-limited chemostat cultures mixed acid product formation was shown to be linked to the growth rate, and homolactic fermentation occurred only in resting cells. Two of the four lactococcal strains investigated with maltose, L. lactis 65.1 and MG1363, showed more pronounced mixed acid product formation during growth than L. lactis ATCC 19435 or IL-1403. In resting cell experiments all four strains exhibited homolactic fermentation. In resting cells the intracellular concentrations of ADP, ATP, and fructose 1,6-bisphosphate were increased and the concentration of Pi was decreased compared with the concentrations in growing cells. Addition of an ionophore (monensin or valinomycin) to resting cultures of L. lactis 65.1 induced mixed acid product formation concomitant with decreases in the ADP, ATP, and fructose 1,6-bisphosphate concentrations. ADP and ATP were shown to inhibit glyceraldehyde-3-phosphate dehydrogenase, lactate dehydrogenase, and alcohol dehydrogenase in vitro. Alcohol dehydrogenase was the most sensitive enzyme and was totally inhibited at an adenine nucleotide concentration of 16 mM, which is close to the sum of the intracellular concentrations of ADP and ATP of resting cells. This inhibition of alcohol dehydrogenase might be partially responsible for the homolactic behavior of resting cells. A hypothesis regarding the level of the ATP-ADP pool as a regulating mechanism for the glycolytic flux and product formation in L. lactis is discussed. PMID:15345435

  17. Presenilin and nicastrin regulate each other and determine amyloid β-peptide production via complex formation

    PubMed Central

    Edbauer, Dieter; Winkler, Edith; Haass, Christian; Steiner, Harald

    2002-01-01

    Amyloid β-peptide (Aβ) is generated by the consecutive cuts of two membrane-bound proteases. β-Secretase cuts at the N terminus of the Aβ domain, whereas γ-secretase mediates the C-terminal cut. Recent evidence suggests that the presenilin (PS) proteins, PS1 and PS2, may be γ-secretases. Because PSs principally exist as high molecular weight protein complexes, biologically active γ-secretases likely require other cofactors such as nicastrin (Nct) for their activities. Here we show that preferentially mature Nct forms a stable complex with PSs. Furthermore, we have down-regulated Nct levels by using a highly specific and efficient RNA interference approach. Very similar to a loss of PS function, down-regulation of Nct levels leads to a massive accumulation of the C-terminal fragments of the β-amyloid precursor protein. In addition, Aβ production was markedly reduced. Strikingly, down-regulation of Nct destabilized PS and strongly lowered levels of the high molecular weight PS1 complex. Interestingly, absence of the PS1 complex in PS1−/− cells was associated with a strong down-regulation of the levels of mature Nct, suggesting that binding to PS is required for trafficking of Nct through the secretory pathway. Based on these findings we conclude that Nct and PS regulate each other and determine γ-secretase function via complex formation. PMID:12048259

  18. β-Adrenergic Receptors Regulate the Acquisition and Consolidation Phases of Aversive Memory Formation Through Distinct, Temporally Regulated Signaling Pathways.

    PubMed

    Schiff, Hillary C; Johansen, Joshua P; Hou, Mian; Bush, David E A; Smith, Emily K; Klein, JoAnna E; LeDoux, Joseph E; Sears, Robert M

    2017-03-01

    Memory formation requires the temporal coordination of molecular events and cellular processes following a learned event. During Pavlovian threat (fear) conditioning (PTC), sensory and neuromodulatory inputs converge on post-synaptic neurons within the lateral nucleus of the amygdala (LA). By activating an intracellular cascade of signaling molecules, these G-protein-coupled neuromodulatory receptors are capable of recruiting a diverse profile of plasticity-related proteins. Here we report that norepinephrine, through its actions on β-adrenergic receptors (βARs), modulates aversive memory formation following PTC through two molecularly and temporally distinct signaling mechanisms. Specifically, using behavioral pharmacology and biochemistry in adult rats, we determined that βAR activity during, but not after PTC training initiates the activation of two plasticity-related targets: AMPA receptors (AMPARs) for memory acquisition and short-term memory and extracellular regulated kinase (ERK) for consolidating the learned association into a long-term memory. These findings reveal that βAR activity during, but not following PTC sets in motion cascading molecular events for the acquisition (AMPARs) and subsequent consolidation (ERK) of learned associations.

  19. Mechanism of interaction of Dictyostelium severin with actin filaments

    PubMed Central

    1982-01-01

    Severin, a 40,000-dalton protein from Dictyostelium that disassembles actin filaments in a Ca2+ -dependent manner, was purified 500-fold to greater than 99% homogeneity by modifications of the procedure reported by Brown, Yamamoto, and Spudich (1982. J. Cell Biol. 93:205-210). Severin has a Stokes radius of 29 A and consists of a single polypeptide chain. It contains a single methionyl and five cysteinyl residues. We studied the action of severin on actin filaments by electron microscopy, viscometry, sedimentation, nanosecond emission anisotropy, and fluorescence energy transfer spectroscopy. Nanosecond emission anisotropy of fluoresence-labeled severin shows that this protein changes its conformation on binding Ca2+. Actin filaments are rapidly fragmented on addition of severin and Ca2+, but severin does not interact with actin filaments in the absence of Ca2+. Fluorescence energy transfer measurements indicate that fragmentation of actin filaments by severin leads to a partial depolymerization (t1/2 approximately equal to 30 s). Depolymerization is followed by exchange of a limited number of subunits in the filament fragments with the disassembled actin pool (t1/2 approximately equal to 5 min). Disassembly and exchange are probably restricted to the ends of the filament fragments since only a few subunits in each fragment participate in the disassembly or exchange process. Steady state hydrolysis of ATP by actin in the presence of Ca2+-severin is maximal at an actin: severin molar ratio of approximately 10:1, which further supports the inference that subunit exchange is limited to the ends of actin filaments. The observation of sequential depolymerization and subunit exchange following the fragmentation of actin by severin suggests that severin may regulate site-specific disassembly and turnover of actin filament arrays in vivo. PMID:6897549

  20. Force Generation, Polymerization Dynamics and Nucleation of Actin Filaments

    NASA Astrophysics Data System (ADS)

    Wang, Ruizhe

    We study force generation and actin filament dynamics using stochastic and deterministic methods. First, we treat force generation of bundled actin filaments by polymerization via molecular-level stochastic simulations. In the widely-used Brownian Ratchet model, actin filaments grow freely whenever the tip-obstacle gap created by thermal fluctuation exceeds the monomer size. We name this model the Perfect Brownian Ratchet (PBR) model. In the PBR model, actin monomer diffusion is treated implicitly. We perform a series of simulations based on the PBR, in which obstacle motion is treated explicitly; in most previous studies, obstacle motion has been treated implicitly. We find that the cooperativity of filaments is generally weak in the PBR model, meaning that more filaments would grow more slowly given the same force per filament. Closed-form formulas are also developed, which match the simulation results. These portable and accurate formulas provide guidance for experiments and upper and lower bounds for theoretical analyses. We also studied a variation of the PBR, called the Diffusing Brownian Ratchet (DBR) model, in which both actin monomer and obstacle diffusion are treated explicitly. We find that the growth rate of multiple filaments is even lower, compared with that in PBR. This finding challenges the widely-accepted PBR assumption and suggests that pushing the study of actin dynamics down to the sub-nanometer level yields new insights. We subsequently used a rate equation approach to model the effect of local depletion of actin monomers on the nucleation of actin filaments on biomimetic beads, and how the effect is regulated by capping protein (CP). We find that near the bead surface, a higher CP concentration increases local actin concentration, which leads to an enhanced activities of actin filaments' nucleation. Our model analysis matches the experimental results and lends support to an important but undervalued hypothesis proposed by Carlier and

  1. Mechanosensing through focal adhesion-anchored intermediate filaments.

    PubMed

    Gregor, Martin; Osmanagic-Myers, Selma; Burgstaller, Gerald; Wolfram, Michael; Fischer, Irmgard; Walko, Gernot; Resch, Guenter P; Jörgl, Almut; Herrmann, Harald; Wiche, Gerhard

    2014-02-01

    Integrin-based mechanotransduction involves a complex focal adhesion (FA)-associated machinery that is able to detect and respond to forces exerted either through components of the extracellular matrix or the intracellular contractile actomyosin network. Here, we show a hitherto unrecognized regulatory role of vimentin intermediate filaments (IFs) in this process. By studying fibroblasts in which vimentin IFs were decoupled from FAs, either because of vimentin deficiency (V0) or loss of vimentin network anchorage due to deficiency in the cytolinker protein plectin (P0), we demonstrate attenuated activation of the major mechanosensor molecule FAK and its downstream targets Src, ERK1/2, and p38, as well as an up-regulation of the compensatory feedback loop acting on RhoA and myosin light chain. In line with these findings, we show strongly reduced FA turnover rates in P0 fibroblasts combined with impaired directional migration, formation of protrusions, and up-regulation of "stretched" high-affinity integrin complexes. By exploiting tension-independent conditions, we were able to mechanistically link these defects to diminished cytoskeletal tension in both P0 and V0 cells. Our data provide important new insights into molecular mechanisms underlying cytoskeleton-regulated mechanosensing, a feature that is fundamental for controlled cell movement and tumor progression.

  2. MAP Kinase-Mediated Negative Regulation of Symbiotic Nodule Formation in Medicago truncatula

    PubMed Central

    Ryu, Hojin; Laffont, Carole; Frugier, Florian; Hwang, Ildoo

    2017-01-01

    Mitogen-activated protein kinase (MAPK) signaling cascades play critical roles in various cellular events in plants, including stress responses, innate immunity, hormone signaling, and cell specificity. MAPK-mediated stress signaling is also known to negatively regulate nitrogen-fixing symbiotic interactions, but the molecular mechanism of the MAPK signaling cascades underlying the symbiotic nodule development remains largely unknown. We show that the MtMKK5-MtMPK3/6 signaling module negatively regulates the early symbiotic nodule formation, probably upstream of ERN1 (ERF Required for Nodulation 1) and NSP1 (Nod factor Signaling Pathway 1) in Medicago truncatula. The overexpression of MtMKK5 stimulated stress and defense signaling pathways but also reduced nodule formation in M. truncatula roots. Conversely, a MAPK specific inhibitor, U0126, enhanced nodule formation and the expression of an early nodulation marker gene, MtNIN. We found that MtMKK5 directly activates MtMPK3/6 by phosphorylating the TEY motif within the activation loop and that the MtMPK3/6 proteins physically interact with the early nodulation-related transcription factors ERN1 and NSP1. These data suggest that the stress signaling-mediated MtMKK5/MtMPK3/6 module suppresses symbiotic nodule development via the action of early nodulation transcription factors. PMID:28152300

  3. Signaling by bone morphogenetic proteins directs formation of an ectodermal signaling center that regulates craniofacial development.

    PubMed

    Foppiano, Silvia; Hu, Diane; Marcucio, Ralph S

    2007-12-01

    We previously described a signaling center, the Frontonasal Ectodermal Zone (FEZ) that regulates growth and patterning of the frontonasal process (FNP). The FEZ is comprised of FNP ectoderm flanking a boundary between Sonic hedgehog (Shh) and Fibroblast growth factor 8 (Fgf8) expression domains. Our objective was to examine BMP signaling during formation of the FEZ. We blocked BMP signaling throughout the FNP prior to FEZ formation by infecting chick embryos at stage 10 (HH10) with a replication-competent avian retrovirus encoding the BMP antagonist Noggin. We assessed gene expression patterns in the FNP 72 h after infection (approximately HH22) and observed that Shh expression was reduced or absent. In the mesenchyme, we observed that Bmp2 transcripts were absent while the Bmp4 expression domain was expanded proximally. In addition to the molecular changes, infected embryos also exhibited facial malformations at 72 and 96 h after infection suggesting that the FEZ did not form. Our data indicate that reduced cell proliferation, but not apoptosis, in the mesenchyme contributed to the phenotype that we observed. Additionally, adding exogenous SHH into the mesenchyme of RCAS-Noggin-infected embryos did not restore Bmp2 and Bmp4 to a normal pattern of expression. These data indicate that BMP signaling mediates interactions between tissues in the FNP that regulate FEZ formation; and that the correct pattern of Bmp2 and Bmp4, but not Bmp7, expression in the FNP mesenchyme requires signaling by the BMP pathway.

  4. Chondroitin 6-Sulfation Regulates Perineuronal Net Formation by Controlling the Stability of Aggrecan.

    PubMed

    Miyata, Shinji; Kitagawa, Hiroshi

    2016-01-01

    Perineuronal nets (PNNs) are lattice-like extracellular matrix structures composed of chondroitin sulfate proteoglycans (CSPGs). The appearance of PNNs parallels the decline of neural plasticity, and disruption of PNNs reactivates neural plasticity in the adult brain. We previously reported that sulfation patterns of chondroitin sulfate (CS) chains on CSPGs influenced the formation of PNNs and neural plasticity. However, the mechanism of PNN formation regulated by CS sulfation remains unknown. Here we found that overexpression of chondroitin 6-sulfotransferase-1 (C6ST-1), which catalyzes 6-sulfation of CS chains, selectively decreased aggrecan, a major CSPG in PNNs, in the aged brain without affecting other PNN components. Both diffuse and PNN-associated aggrecans were reduced by overexpression of C6ST-1. C6ST-1 increased 6-sulfation in both the repeating disaccharide region and linkage region of CS chains. Overexpression of 6-sulfation primarily impaired accumulation of aggrecan in PNNs, whereas condensation of other PNN components was not affected. Finally, we found that increased 6-sulfation accelerated proteolysis of aggrecan by a disintegrin and metalloproteinase domain with thrombospondin motif (ADAMTS) protease. Taken together, our results indicate that sulfation patterns of CS chains on aggrecan influenced the stability of the CSPG, thereby regulating formation of PNNs and neural plasticity.

  5. Hydrogen sulfide is a novel gasotransmitter with pivotal role in regulating lateral root formation in plants

    PubMed Central

    Li, Yan-Jun; Shi, Zhi-Qi; Gan, Li-Jun; Chen, Jian

    2014-01-01

    Hydrogen sulfide (H2S), the third gasotransmitter after nitric oxide (NO) and carbon monoxide (CO), is a critical neuromodulator in the pathogenesis of various diseases from neurodegenerative diseases to diabetes or heart failure. The crosstalk between NO and H2S has been well established in mammalian physiology. In planta, NO is demonstrated to regulate lateral root formation by acting downstream of auxin. The recent reports revealed that H2S is a novel inducer of lateral root (LR) formation by stimulating the expression of cell cycle regulatory genes (CCRGs), acting similarly with NO, CO, and IAA. Interestingly, during the initiation of lateral root primordia, IAA is a potent inducer of endogenous H2S and CO, which is produced by L-cysteine desulfhydrase (LCD) and heme oxygenase-1 (HO-1), respectively. The increasing evidences suggest that H2S-promoted LR growth is dependent on the endogenous production of CO. In addition, our results indicate that the H2S signaling in the regulation of LR formation can be associated to NO and Ca2+. In this addendum, we advanced a proposed schematic model for H2S-mediated signaling pathway of plant LR development. PMID:24832131

  6. A Genome-Wide Association Study for Regulators of Micronucleus Formation in Mice

    PubMed Central

    McIntyre, Rebecca E.; Nicod, Jérôme; Robles-Espinoza, Carla Daniela; Maciejowski, John; Cai, Na; Hill, Jennifer; Verstraten, Ruth; Iyer, Vivek; Rust, Alistair G.; Balmus, Gabriel; Mott, Richard; Flint, Jonathan; Adams, David J.

    2016-01-01

    In mammals the regulation of genomic instability plays a key role in tumor suppression and also controls genome plasticity, which is important for recombination during the processes of immunity and meiosis. Most studies to identify regulators of genomic instability have been performed in cells in culture or in systems that report on gross rearrangements of the genome, yet subtle differences in the level of genomic instability can contribute to whole organism phenotypes such as tumor predisposition. Here we performed a genome-wide association study in a population of 1379 outbred Crl:CFW(SW)-US_P08 mice to dissect the genetic landscape of micronucleus formation, a biomarker of chromosomal breaks, whole chromosome loss, and extranuclear DNA. Variation in micronucleus levels is a complex trait with a genome-wide heritability of 53.1%. We identify seven loci influencing micronucleus formation (false discovery rate <5%), and define candidate genes at each locus. Intriguingly at several loci we find evidence for sexual dimorphism in micronucleus formation, with a locus on chromosome 11 being specific to males. PMID:27233670

  7. A Genome-Wide Association Study for Regulators of Micronucleus Formation in Mice.

    PubMed

    McIntyre, Rebecca E; Nicod, Jérôme; Robles-Espinoza, Carla Daniela; Maciejowski, John; Cai, Na; Hill, Jennifer; Verstraten, Ruth; Iyer, Vivek; Rust, Alistair G; Balmus, Gabriel; Mott, Richard; Flint, Jonathan; Adams, David J

    2016-08-09

    In mammals the regulation of genomic instability plays a key role in tumor suppression and also controls genome plasticity, which is important for recombination during the processes of immunity and meiosis. Most studies to identify regulators of genomic instability have been performed in cells in culture or in systems that report on gross rearrangements of the genome, yet subtle differences in the level of genomic instability can contribute to whole organism phenotypes such as tumor predisposition. Here we performed a genome-wide association study in a population of 1379 outbred Crl:CFW(SW)-US_P08 mice to dissect the genetic landscape of micronucleus formation, a biomarker of chromosomal breaks, whole chromosome loss, and extranuclear DNA. Variation in micronucleus levels is a complex trait with a genome-wide heritability of 53.1%. We identify seven loci influencing micronucleus formation (false discovery rate <5%), and define candidate genes at each locus. Intriguingly at several loci we find evidence for sexual dimorphism in micronucleus formation, with a locus on chromosome 11 being specific to males.

  8. Membrane-Associated Molecules Regulate the Formation of Layer-Specific Cortical Circuits

    NASA Astrophysics Data System (ADS)

    Castellani, Valerie; Bolz, Jurgen

    1997-06-01

    The columnar organization of the mammalian neocortex is based on radially oriented axon collaterals which precisely link cells from distinct cortical layers. During development, these interlaminar connections are specific from their initial outgrowth: collaterals form only in the target layers and there are no transient axonal collaterals in the nontarget layers. To examine whether positional cues within individual cortical layers regulate the laminar specificity of collateral formation, explants of cells destined for different cortical layers were cultured on membranes prepared from target and nontarget layers. Axonal growth and branching were examined on homogeneous membrane substrates and on alternating stripes of membranes from different layers. Results show that axons branch preferentially on membrane substrates from those layers that they would target in vivo. In addition, when cortical axons were given a choice to grow on membranes from either their target or their nontarget layer, they exhibited a clear preference for the target layers. This indicates that membrane-associated cues confined to individual layers regulate the formation of collaterals of cortical axons and restrict their growth to their target layers. Heat inactivation of membranes from target layers resulted in reduced axonal branching. The same manipulation of membranes from nontarget layers increased axonal branching for one population of cortical neurons. Taken together, these results suggest that membrane-associated molecules confined to individual layers induce and prevent the formation of axon collaterals in distinct populations of cortical neurons. Thus, the expression of layer-specific cues provides important constraints for the remodeling of local circuits during cortical development.

  9. LMO4 functions as a negative regulator of sensory organ formation in the mammalian cochlea.

    PubMed

    Deng, Min; Luo, Xiong-jian; Pan, Ling; Yang, Hua; Xie, Xiaoling; Liang, Guoqing; Huang, Liang; Hu, Fang; Kiernan, Amy E; Gan, Lin

    2014-07-23

    In mammals, formation of the auditory sensory organ (the organ of Corti) is restricted to a specialized area of the cochlea. However, the molecular mechanisms limiting sensory formation to this discrete region in the ventral cochlear duct are not well understood, nor is it known whether other regions of the cochlea have the competence to form the organ of Corti. Here we identify LMO4, a LIM-domain-only nuclear protein, as a negative regulator of sensory organ formation in the cochlea. Inactivation of Lmo4 in mice leads to an ectopic organ of Corti (eOC) located in the lateral cochlea. The eOC retains the features of the native organ, including inner and outer hair cells, supporting cells, and other nonsensory specialized cell types. However, the eOC shows an orientation opposite to the native organ, such that the eOC appears as a mirror-image duplication to the native organ of Corti. These data demonstrate a novel sensory competent region in the lateral cochlear duct that is regulated by LMO4 and may be amenable to therapeutic manipulation.

  10. Centriolar SAS-7 acts upstream of SPD-2 to regulate centriole assembly and pericentriolar material formation.

    PubMed

    Sugioka, Kenji; Hamill, Danielle R; Lowry, Joshua B; McNeely, Marie E; Enrick, Molly; Richter, Alyssa C; Kiebler, Lauren E; Priess, James R; Bowerman, Bruce

    2017-01-16

    The centriole/basal body is a eukaryotic organelle that plays essential roles in cell division and signaling. Among five known core centriole proteins, SPD-2/Cep192 is the first recruited to the site of daughter centriole formation and regulates the centriolar localization of the other components in C. elegans and in humans. However, the molecular basis for SPD-2 centriolar localization remains unknown. Here we describe a new centriole component, the coiled-coil protein SAS-7, as a regulator of centriole duplication, assembly and elongation. We also show that SAS-7 binds SPD-2 and regulates SPD-2 centriolar recruitment, while SAS-7 centriolar localization is SPD-2-independent. Furthermore, pericentriolar material (PCM) formation is abnormal in sas-7 mutants, and the PCM-dependent induction of cell polarity that defines the anterior-posterior body axis frequently fails. We conclude that SAS-7 functions at the earliest step in centriole duplication yet identified and plays important roles in the orchestration of centriole and PCM assembly.

  11. Centriolar SAS-7 acts upstream of SPD-2 to regulate centriole assembly and pericentriolar material formation

    PubMed Central

    Sugioka, Kenji; Hamill, Danielle R; Lowry, Joshua B; McNeely, Marie E; Enrick, Molly; Richter, Alyssa C; Kiebler, Lauren E; Priess, James R; Bowerman, Bruce

    2017-01-01

    The centriole/basal body is a eukaryotic organelle that plays essential roles in cell division and signaling. Among five known core centriole proteins, SPD-2/Cep192 is the first recruited to the site of daughter centriole formation and regulates the centriolar localization of the other components in C. elegans and in humans. However, the molecular basis for SPD-2 centriolar localization remains unknown. Here, we describe a new centriole component, the coiled-coil protein SAS-7, as a regulator of centriole duplication, assembly and elongation. Intriguingly, our genetic data suggest that SAS-7 is required for daughter centrioles to become competent for duplication, and for mother centrioles to maintain this competence. We also show that SAS-7 binds SPD-2 and regulates SPD-2 centriolar recruitment, while SAS-7 centriolar localization is SPD-2-independent. Furthermore, pericentriolar material (PCM) formation is abnormal in sas-7 mutants, and the PCM-dependent induction of cell polarity that defines the anterior-posterior body axis frequently fails. We conclude that SAS-7 functions at the earliest step in centriole duplication yet identified and plays important roles in the orchestration of centriole and PCM assembly. DOI: http://dx.doi.org/10.7554/eLife.20353.001 PMID:28092264

  12. Wood Formation in Trees Is Increased by Manipulating PXY-Regulated Cell Division.

    PubMed

    Etchells, J Peter; Mishra, Laxmi S; Kumar, Manoj; Campbell, Liam; Turner, Simon R

    2015-04-20

    The woody tissue of trees is composed of xylem cells that arise from divisions of stem cells within the cambial meristem. The rate of xylem cell formation is dependent upon the rate of cell division within the cambium and is controlled by both genetic and environmental factors. In the annual plant Arabidopsis, signaling between a peptide ligand CLE41 and a receptor kinase PXY controls cambial cell divisions; however, the pathway regulating secondary growth in trees has not been identified. Here, we show that an aspen receptor kinase PttPXY and its peptide ligand PttCLE41 are functional orthologs and act to control a multifunctional pathway that regulates both the rate of cambial cell division and woody tissue organization. Ectopic overexpression of PttPXY and PttCLE41 genes in hybrid aspen resulted in vascular tissue abnormalities and poor plant growth. In contrast, precise tissue-specific overexpression generated trees that exhibited a 2-fold increase in the rate of wood formation, were taller, and possessed larger leaves compared to the controls. Our results demonstrate that the PXY-CLE pathway has evolved to regulate secondary growth and manipulating this pathway can result in dramatically increased tree growth and productivity.

  13. Proteomic Analysis of Fetal Ovaries Reveals That Primordial Follicle Formation and Transition Are Differentially Regulated

    PubMed Central

    Xu, Mengmeng; Che, Long; Yang, Zhenguo; Zhang, Pan; Shi, Jiankai; Li, Jian; Lin, Yan; Fang, Zhengfeng; Che, Lianqiang; Feng, Bin; Wu, De

    2017-01-01

    Primordial follicle formation represents a critical phase of the initiation of embryonic reproductive organ development, while the primordial follicle transition into primary follicle determines whether oestrus or ovulation will occur in female animals. To identify molecular mechanism of new proteins which are involved in ovarian development, we employed 2D-DIGE to compare the protein expression profiles of primordial follicles and primary follicles of fetal ovaries in pigs. Fetal ovaries were collected at distinct time-points of the gestation cycle (g55 and g90). The identified proteins at the g55 time-point are mainly involved in the development of anatomical structures [reticulocalbin-1 (RCN1), reticulocalbin-3 (RCN3)], cell differentiation (actin), and stress response [heterogeneous nuclear ribonucleoprotein K (HNRNPK)]. Meanwhile, at the g90 stage, the isolated proteins with altered expression levels were mainly associated with cell proliferation [major vault protein (MVP)] and stress response [heat shock-related 70 kDa protein 2 (HSPA2)]. In conclusion, our work revealed that primordial follicle formation is regulated by RCN1, RCN3, actin, and HNRNPK, while the primordial follicle transformation to primary follicle is regulated by MVP and HSPA2. Therefore, our results provide further information for the prospective understanding of the molecular mechanism(s) involved in the regulation of the ovarian follicle development. PMID:28265575

  14. Epigenetic gene regulation in the adult mammalian brain: multiple roles in memory formation.

    PubMed

    Lubin, Farah D

    2011-07-01

    Brain-derived neurotrophic factor (bdnf) is one of numerous gene products necessary for long-term memory formation and dysregulation of bdnf has been implicated in the pathogenesis of cognitive and mental disorders. Recent work indicates that epigenetic-regulatory mechanisms including the markings of histone proteins and associated DNA remain labile throughout the life-span and represent an attractive molecular process contributing to gene regulation in the brain. In this review, important information will be discussed on epigenetics as a set of newly identified dynamic transcriptional mechanisms serving to regulate gene expression changes in the adult brain with particular emphasis on bdnf transcriptional readout in learning and memory formation. This review will also highlight evidence for the role of epigenetics in aberrant bdnf gene regulation in the pathogenesis of cognitive dysfunction associated with seizure disorders, Rett syndrome, Schizophrenia, and Alzheimer's disease. Such research offers novel concepts for understanding epigenetic transcriptional mechanisms subserving adult cognition and mental health, and furthermore promises novel avenues for therapeutic approach in the clinic.

  15. F actin bundles in Drosophila bristles. I. Two filament cross-links are involved in bundling

    PubMed Central

    1995-01-01

    Transverse sections though Drosophila bristles reveal 7-11 nearly round, plasma membrane-associated bundles of actin filaments. These filaments are hexagonally packed and in a longitudinal section they show a 12-nm periodicity in both the 1.1 and 1.0 views. From earlier studies this periodicity is attributable to cross-links and indicates that the filaments are maximally cross-linked, singed mutants also have 7-11 bundles, but the bundles are smaller, flattened, and the filaments within the bundles are randomly packed (not hexagonal); no periodicity can be detected in longitudinal sections. Another mutant, forked (f36a), also has 7-11 bundles but even though the bundles are very small, the filaments within them are hexagonally packed and display a 12-nm periodicity in longitudinal section. The singed-forked double mutant lacks filament bundles. Thus there are at least two species of cross-links between adjacent actin filaments. Hints of why two species of cross-links are necessary can be gleaned by studying bristle formation. Bristles sprout with only microtubules within them. A little later in development actin filaments appear. At early stages the filaments in the bundles are randomly packed. Later the filaments in the bundles become hexagonally packed and maximally cross-linked. We consider that the forked proteins may be necessary early in development to tie the filaments together in a bundle so that they can be subsequently zippered together by fascin (the singed gene product). PMID:7622563

  16. Theory of electron current filamentation instability and ion density filamentation in the early development of a DPF discharge

    SciTech Connect

    Guillory, J.; Rose, D. V.; Lerner, E. J.

    2009-01-21

    Two-dimensional simulations of the initial stages of plasma formation in a dense plasma focus show the formation, in a few tens of nanoseconds, of a dense layer of plasma (n{sub e}{approx}10{sup 18} cm{sup -3},T{sub e}{approx}3 eV) in a thin layer surrounding the insulator-covered central anode of the focus device, and carrying axially-directed current at rather high current density.Earlier work on the filamentation of dense cathode plasma in high-power diodes seems to indicate that the anode plasma current layer in a dense plasma focus (DPF) device could be subject to the same instability, creating a growth of axially-directed filaments in the current density. The growth rate for resistive-thermal-driven filamentation, e.g. at 30 torr and {approx}3 eV electron temperature, exceeds the that due to non-thermal current (JxB) driving, and is determined by electron dynamics, so its evolution is quicker than the response-time of the ions.Nonetheless, with such a growing current-density perturbation as a seed and its increasing rippling of the azimuthal magnetic field as a driver, the ions will eventually take part in the azimuthal bunching, forming filaments in the ion density as well. The resistive-thermal-driven filamentation fields thus serve to 'hurry-up' the development of ion density filamentation, as shown approximately in the work presented here. This theory predicts, for light ions, a relatively early ({<=}250 ns) development of visible filaments along the anode, perhaps even before the main rundown phase of the focus plasma motion, and these filaments may persist during the 'liftoff' phase of the current layer to form the rundown phase of the plasma front. This work is supported by Larwenceville Plasma Physics.

  17. Cloning and characterization of a type-A response regulator differentially expressed during adventitious shoot formation in Pinus pinea L.

    PubMed

    Cortizo, M; Alvarez, J M; Rodríguez, A; Fernández, B; Ordás, R J

    2010-08-15

    Type-A response regulators play an important role in cytokinin-induced adventitious shoot formation, acting as negative regulators of cytokinin signal transduction. In this work, we obtained the full-length cDNA clone of a type-A response regulator from the conifer Pinus pinea, designated PipiRR1. The derived peptide sequence showed all the characteristic motifs found in angiosperms. Gene expression analysis showed that the gene was differentially expressed during adventitious shoot formation in P. pinea cotyledons, suggesting that PipiRR1 may play a role in caulogenesis in conifers. This is the first type-A response regulator identified in gymnosperms.

  18. Role of WNT16 in the regulation of periosteal bone formation in female mice.

    PubMed

    Wergedal, Jon E; Kesavan, Chandrasekhar; Brommage, Robert; Das, Subhashri; Mohan, Subburaman

    2015-03-01

    In this study, we evaluated the role of WNT16 in regulating bone size, an important determinant of bone strength. Mice with targeted disruption of the Wnt16 gene exhibited a 24% reduction in tibia cross-sectional area at 12 weeks of age compared with that of littermate wild-type (WT) mice. Histomorphometric studies revealed that the periosteal bone formation rate and mineral apposition rate were reduced (P < .05) by 55% and 32%, respectively, in Wnt16 knockout (KO) vs WT mice at 12 weeks of age. In contrast, the periosteal tartrate resistant acid phosphatase-labeled surface was increased by 20% in the KO mice. Because mechanical strain is an important physiological regulator of periosteal bone formation (BF), we determined whether mechanical loading-induced periosteal BF is compromised in Wnt16 KO mice. Application of 4800-μe strain to the right tibia using a 4-point bending loading method for 2 weeks (2-Hz frequency, 36 cycles per day, 6 days/wk) produced a significant increase in cross-sectional area (11% above that of the unloaded left tibia, P < .05, n = 6) in the WT but not in the KO mice (-0.2% change). Histomorphometric analyses revealed increases in the periosteal bone formation rate and mineral apposition rate in the loaded bones of WT but not KO mice. Wnt16 KO mice showed significant (20%-70%) reductions in the expression levels of markers of canonical (β-catenin and Axin2) but not noncanonical (Nfatc1 and Tnnt2) WNT signaling in the periosteum at 5 weeks of age. Our findings suggest that WNT16 acting via canonical WNT signaling regulates mechanical strain-induced periosteal BF and bone size.

  19. RALFL34 regulates formative cell divisions in Arabidopsis pericycle during lateral root initiation.

    PubMed

    Murphy, Evan; Vu, Lam Dai; Van den Broeck, Lisa; Lin, Zhefeng; Ramakrishna, Priya; van de Cotte, Brigitte; Gaudinier, Allison; Goh, Tatsuaki; Slane, Daniel; Beeckman, Tom; Inzé, Dirk; Brady, Siobhan M; Fukaki, Hidehiro; De Smet, Ive

    2016-08-01

    In plants, many signalling molecules, such as phytohormones, miRNAs, transcription factors, and small signalling peptides, drive growth and development. However, very few small signalling peptides have been shown to be necessary for lateral root development. Here, we describe the role of the peptide RALFL34 during early events in lateral root development, and demonstrate its specific importance in orchestrating formative cell divisions in the pericycle. Our results further suggest that this small signalling peptide acts on the transcriptional cascade leading to a new lateral root upstream of GATA23, an important player in lateral root formation. In addition, we describe a role for ETHYLENE RESPONSE FACTORs (ERFs) in regulating RALFL34 expression. Taken together, we put forward RALFL34 as a new, important player in lateral root initiation.

  20. Electrical coupling regulates layer 1 interneuron microcircuit formation in the neocortex

    PubMed Central

    Yao, Xing-Hua; Wang, Min; He, Xiang-Nan; He, Fei; Zhang, Shu-Qing; Lu, Wenlian; Qiu, Zi-Long; Yu, Yong-Chun

    2016-01-01

    The coexistence of electrical and chemical synapses among interneurons is essential for interneuron function in the neocortex. However, it remains largely unclear whether electrical coupling between interneurons influences chemical synapse formation and microcircuit assembly during development. Here, we show that electrical and GABAergic chemical connections robustly develop between interneurons in neocortical layer 1 over a similar time course. Electrical coupling promotes action potential generation and synchronous firing between layer 1 interneurons. Furthermore, electrically coupled interneurons exhibit strong GABA-A receptor-mediated synchronous synaptic activity. Disruption of electrical coupling leads to a loss of bidirectional, but not unidirectional, GABAergic connections. Moreover, a reduction in electrical coupling induces an increase in excitatory synaptic inputs to layer 1 interneurons. Together, these findings strongly suggest that electrical coupling between neocortical interneurons plays a critical role in regulating chemical synapse development and precise formation of circuits. PMID:27510304

  1. Regulation of gemma formation in the copper moss Scopelophila cataractae by environmental copper concentrations.

    PubMed

    Nomura, Toshihisa; Hasezawa, Seiichiro

    2011-09-01

    Considerable attention has recently been focused on the use of hyperaccumulator plants for the phytoremediation of soils contaminated with heavy metals. The moss, Scopelophila cataractae (Mitt.) Broth., is a typical hyperaccumulator that is usually observed only in copper-rich environments and which accumulates high concentrations of copper in its tissues. However, many of the physiological processes and mechanisms for metal hyperaccumulation in S. cataractae remain unknown. To address this issue, we examined the mechanisms regulating gemma formation, which is considered the main strategy by which S. cataractae relocates to new copper-rich areas. From this study we found that treatment of S. cataractae with high concentrations of copper suppressed gemma formation but promoted protonemal growth. The suppressive effect was not observed by treatment with heavy metals other than copper. These results suggest the importance of copper-sensitive asexual reproduction in the unique life strategy of the copper moss, S. cataractae.

  2. Regulation of ceramide channel formation and disassembly: Insights on the initiation of apoptosis

    PubMed Central

    Abou-Ghali, Majdouline; Stiban, Johnny

    2015-01-01

    Sphingolipid research has surged in the past two decades and has produced a wide variety of evidence supporting the role of this class of molecules in mediating cellular growth, differentiation, senescence, and apoptosis. Ceramides are a subgroup of sphingolipids (SLs) that are directly involved in the process of initiation of apoptosis. We, and others, have recently shown that ceramides are capable of the formation of protein-permeable channels in mitochondrial outer membranes under physiological conditions. These pores are indeed good candidates for the pathway of release of pro-apoptotic proteins from the mitochondrial intermembrane space (IMS) into the cytosol to initiate intrinsic apoptosis. Here, we review recent findings on the regulation of ceramide channel formation and disassembly, highlighting possible implications on the initiation of the intrinsic apoptotic pathway. PMID:26587005

  3. RALFL34 regulates formative cell divisions in Arabidopsis pericycle during lateral root initiation

    PubMed Central

    Murphy, Evan; Vu, Lam Dai; Van den Broeck, Lisa; Lin, Zhefeng; Ramakrishna, Priya; van de Cotte, Brigitte; Gaudinier, Allison; Goh, Tatsuaki; Slane, Daniel; Beeckman, Tom; Inzé, Dirk; Brady, Siobhan M.; Fukaki, Hidehiro; De Smet, Ive

    2016-01-01

    In plants, many signalling molecules, such as phytohormones, miRNAs, transcription factors, and small signalling peptides, drive growth and development. However, very few small signalling peptides have been shown to be necessary for lateral root development. Here, we describe the role of the peptide RALFL34 during early events in lateral root development, and demonstrate its specific importance in orchestrating formative cell divisions in the pericycle. Our results further suggest that this small signalling peptide acts on the transcriptional cascade leading to a new lateral root upstream of GATA23, an important player in lateral root formation. In addition, we describe a role for ETHYLENE RESPONSE FACTORs (ERFs) in regulating RALFL34 expression. Taken together, we put forward RALFL34 as a new, important player in lateral root initiation. PMID:27521602

  4. Self-regulating galaxy formation. Part 1: HII disk and Lyman alpha pressure

    NASA Technical Reports Server (NTRS)

    Cox, D. P.

    1983-01-01

    Assuming a simple but physically based prototype for behavior of interstellar material during formation of a disk galaxy, coupled with the lowest order description of infall, a scenario is developed for self-regulated disk galaxy formation. Radiation pressure, particularly that of Lyman depha (from fluorescence conversion Lyman continuum), is an essential component, maintaining an inflated disk and stopping infall when only a small fraction of the overall perturbation has joined the disk. The resulting galaxies consist of a two dimensional family whose typical scales and surface density are expressable in terms of fundamental constants. The model leads naturally to galaxies with a rich circumgalactic environment and flat rotation curves (but is weak in its analysis of the subsequent evolution of halo material).

  5. Self-regulation of charged defect compensation and formation energy pinning in semiconductors

    PubMed Central

    Yang, Ji-Hui; Yin, Wan-Jian; Park, Ji-Sang; Wei, Su-Huai

    2015-01-01

    Current theoretical analyses of defect properties without solving the detailed balance equations often estimate Fermi-level pinning position by omitting free carriers and assume defect concentrations can be always tuned by atomic chemical potentials. This could be misleading in some circumstance. Here we clarify that: (1) Because the Fermi-level pinning is determined not only by defect states but also by free carriers from band-edge states, band-edge states should be treated explicitly in the same footing as the defect states in practice; (2) defect formation energy, thus defect density, could be pinned and independent on atomic chemical potentials due to the entanglement of atomic chemical potentials and Fermi energy, in contrast to the usual expectation that defect formation energy can always be tuned by varying the atomic chemical potentials; and (3) the charged defect compensation behavior, i.e., most of donors are compensated by acceptors or vice versa, is self-regulated when defect formation energies are pinned. The last two phenomena are more dominant in wide-gap semiconductors or when the defect formation energies are small. Using NaCl and CH3NH3PbI3 as examples, we illustrate these unexpected behaviors. Our analysis thus provides new insights that enrich the understanding of the defect physics in semiconductors and insulators. PMID:26584670

  6. An XIST-related small RNA regulates KRAS G-quadruplex formation beyond X-inactivation.

    PubMed

    Chang, Yuli C; Chiu, Chien-Chih; Yuo, Chung-Yee; Chan, Wen-Ling; Chang, Ya-Sian; Chang, Wen-Hsin; Wu, Shou-Mei; Chou, Han-Lin; Liu, Ta-Chih; Lu, Chi-Yu; Yang, Wen-Kuang; Chang, Jan-Gowth

    2016-12-27

    X-inactive-specific transcript (XIST), a long non-coding RNA, is essential for the initiation of X-chromosome inactivation. However, little is known about other roles of XIST in the physiological process in eukaryotic cells. In this study, the bioinformatics approaches revealed XIST could be processed into a small non-coding RNA XPi2. The XPi2 RNA was confirmed by a northern blot assay; its expression was gender-independent, suggesting the role of XPi2 was beyond X-chromosome inactivation. The pull-down assay combined with LC-MS-MS identified two XPi2-associated proteins, nucleolin and hnRNP A1, connected to the formation of G-quadruplex. Moreover, the microarray data showed the knockdown of XPi2 down-regulated the KRAS pathway. Consistently, we tested the expression of ten genes, including KRAS, which was correlated with a G-quadruplex formation and found the knockdown of XPi2 caused a dramatic decrease in the transcription level of KRAS among the ten genes. The results of CD/NMR assay also supported the interaction of XPi2 and the polypurine-polypyrimidine element of KRAS. Accordingly, XPi2 may stimulate the KRAS expression by attenuating G-quadruplex formation. Our present work sheds light on the novel role of small RNA XPi2 in modulating the G-quadruplex formation which may play some essential roles in the KRAS- associated carcinogenesis.

  7. An XIST-related small RNA regulates KRAS G-quadruplex formation beyond X-inactivation

    PubMed Central

    Chang, Yuli C.; Chiu, Chien-Chih; Yuo, Chung-Yee; Chan, Wen-Ling; Chang, Ya-Sian; Chang, Wen-Hsin; Wu, Shou-Mei; Chou, Han-Lin; Liu, Ta-Chih; Lu, Chi-Yu; Yang, Wen-Kuang; Chang, Jan-Gowth

    2016-01-01

    X-inactive-specific transcript (XIST), a long non-coding RNA, is essential for the initiation of X-chromosome inactivation. However, little is known about other roles of XIST in the physiological process in eukaryotic cells. In this study, the bioinformatics approaches revealed XIST could be processed into a small non-coding RNA XPi2. The XPi2 RNA was confirmed by a northern blot assay; its expression was gender-independent, suggesting the role of XPi2 was beyond X-chromosome inactivation. The pull-down assay combined with LC-MS-MS identified two XPi2-associated proteins, nucleolin and hnRNP A1, connected to the formation of G-quadruplex. Moreover, the microarray data showed the knockdown of XPi2 down-regulated the KRAS pathway. Consistently, we tested the expression of ten genes, including KRAS, which was correlated with a G-quadruplex formation and found the knockdown of XPi2 caused a dramatic decrease in the transcription level of KRAS among the ten genes. The results of CD/NMR assay also supported the interaction of XPi2 and the polypurine-polypyrimidine element of KRAS. Accordingly, XPi2 may stimulate the KRAS expression by attenuating G-quadruplex formation. Our present work sheds light on the novel role of small RNA XPi2 in modulating the G-quadruplex formation which may play some essential roles in the KRAS- associated carcinogenesis. PMID:27880931

  8. Self-regulation of charged defect compensation and formation energy pinning in semiconductors

    NASA Astrophysics Data System (ADS)

    Yang, Ji-Hui; Yin, Wan-Jian; Park, Ji-Sang; Wei, Su-Huai

    2015-11-01

    Current theoretical analyses of defect properties without solving the detailed balance equations often estimate Fermi-level pinning position by omitting free carriers and assume defect concentrations can be always tuned by atomic chemical potentials. This could be misleading in some circumstance. Here we clarify that: (1) Because the Fermi-level pinning is determined not only by defect states but also by free carriers from band-edge states, band-edge states should be treated explicitly in the same footing as the defect states in practice; (2) defect formation energy, thus defect density, could be pinned and independent on atomic chemical potentials due to the entanglement of atomic chemical potentials and Fermi energy, in contrast to the usual expectation that defect formation energy can always be tuned by varying the atomic chemical potentials; and (3) the charged defect compensation behavior, i.e., most of donors are compensated by acceptors or vice versa, is self-regulated when defect formation energies are pinned. The last two phenomena are more dominant in wide-gap semiconductors or when the defect formation energies are small. Using NaCl and CH3NH3PbI3 as examples, we illustrate these unexpected behaviors. Our analysis thus provides new insights that enrich the understanding of the defect physics in semiconductors and insulators.

  9. Self-regulation of charged defect compensation and formation energy pinning in semiconductors

    SciTech Connect

    Yang, Ji -Hui; Yin, Wan -Jian; Park, Ji -Sang; Wei, Su -Huai

    2015-11-20

    Current theoretical analyses of defect properties without solving the detailed balance equations often estimate Fermi-level pinning position by omitting free carriers and assume defect concentrations can be always tuned by atomic chemical potentials. This could be misleading in some circumstance. Here we clarify that: (1) Because the Fermi-level pinning is determined not only by defect states but also by free carriers from band-edge states, band-edge states should be treated explicitly in the same footing as the defect states in practice; (2) defect formation energy, thus defect density, could be pinned and independent on atomic chemical potentials due to the entanglement of atomic chemical potentials and Fermi energy, in contrast to the usual expectation that defect formation energy can always be tuned by varying the atomic chemical potentials; and (3) the charged defect compensation behavior, i.e., most of donors are compensated by acceptors or vice versa, is self-regulated when defect formation energies are pinned. The last two phenomena are more dominant in wide-gap semiconductors or when the defect formation energies are small. Using NaCl and CH3NH3PbI3 as examples, we illustrate these unexpected behaviors. Our analysis thus provides new insights that enrich the understanding of the defect physics in semiconductors and insulators.

  10. Self-regulation of charged defect compensation and formation energy pinning in semiconductors

    DOE PAGES

    Yang, Ji -Hui; Yin, Wan -Jian; Park, Ji -Sang; ...

    2015-11-20

    Current theoretical analyses of defect properties without solving the detailed balance equations often estimate Fermi-level pinning position by omitting free carriers and assume defect concentrations can be always tuned by atomic chemical potentials. This could be misleading in some circumstance. Here we clarify that: (1) Because the Fermi-level pinning is determined not only by defect states but also by free carriers from band-edge states, band-edge states should be treated explicitly in the same footing as the defect states in practice; (2) defect formation energy, thus defect density, could be pinned and independent on atomic chemical potentials due to the entanglementmore » of atomic chemical potentials and Fermi energy, in contrast to the usual expectation that defect formation energy can always be tuned by varying the atomic chemical potentials; and (3) the charged defect compensation behavior, i.e., most of donors are compensated by acceptors or vice versa, is self-regulated when defect formation energies are pinned. The last two phenomena are more dominant in wide-gap semiconductors or when the defect formation energies are small. Using NaCl and CH3NH3PbI3 as examples, we illustrate these unexpected behaviors. Our analysis thus provides new insights that enrich the understanding of the defect physics in semiconductors and insulators.« less

  11. Targeting P38 Pathway Regulates Bony Formation via MSC Recruitment during Mandibular Distraction Osteogenesis in Rats

    PubMed Central

    Yang, Zi-hui; Wu, Bao-lei; Ye, Chen; Jia, Sen; Yang, Xin-jie; Hou, Rui; Lei, De-lin; Wang, Lei

    2016-01-01

    Distraction osteogenesis (DO) is a widely used self-tissue engineering. However, complications and discomfort due to the long treatment period are still the bottleneck of DO. Novel strategies to accelerate bone formation in DO are still needed. P38 is capable of regulating the osteogenic differentiation of both mesenchymal stem cells (MSCs) and osteoblasts, which are crucial to bone regeneration. However, it is not clear whether targeting p38 could regulate bony formation in DO. The purpose of the current work was to investigate the effects of local application of either p38 agonist anisomycin or p38 inhibitor SB203580 in a rat model of DO. 30 adult rats were randomly divided into 3 groups: (A) rats injected with DMSO served as the control group; (B) rats injected with p38 agonist anisomycin; (C) rats injected with p38 inhibitor SB203580. All the rats were subjected to mandibular distraction and the injection was performed daily during this period. The distracted mandibles were harvested on days 15 and 30 after surgery and subjected to the following analysis. Micro-computed tomography and histological evaluation results showed that local application of p38 agonist anisomycin increased new bone formation in DO, whereas p38 inhibitor SB203580 decreased it. Immunohistochemical analysis suggested that anisomycin promoted MSC recruitment in the distraction gap. In conclusion, this study demonstrated that local application of p38 agonist anisomycin can increase new bone formation during DO. This study may lead to a novel cell-based strategy for the improvement of bone regeneration. PMID:27766028

  12. Characterization of Key Helicobacter pylori Regulators Identifies a Role for ArsRS in Biofilm Formation

    PubMed Central

    Servetas, Stephanie L.; Carpenter, Beth M.; Haley, Kathryn P.; Gilbreath, Jeremy J.; Gaddy, Jennifer A.

    2016-01-01

    ABSTRACT Helicobacter pylori must be able to rapidly respond to fluctuating conditions within the stomach. Despite this need for constant adaptation, H. pylori encodes few regulatory proteins. Of the identified regulators, the ferric uptake regulator (Fur), the nickel response regulator (NikR), and the two-component acid response system (ArsRS) are each paramount to the success of this pathogen. While numerous studies have individually examined these regulatory proteins, little is known about their combined effect. Therefore, we constructed a series of isogenic mutant strains that contained all possible single, double, and triple regulatory mutations in Fur, NikR, and ArsS. A growth curve analysis revealed minor variation in growth kinetics across the strains; these were most pronounced in the triple mutant and in strains lacking ArsS. Visual analysis showed that strains lacking ArsS formed large aggregates and a biofilm-like matrix at the air-liquid interface. Biofilm quantification using crystal violet assays and visualization via scanning electron microscopy (SEM) showed that all strains lacking ArsS or containing a nonphosphorylatable form of ArsR (ArsR-D52N mutant) formed significantly more biofilm than the wild-type strain. Molecular characterization of biofilm formation showed that strains containing mutations in the ArsRS pathway displayed increased levels of cell aggregation and adherence, both of which are key to biofilm development. Furthermore, SEM analysis revealed prevalent coccoid cells and extracellular matrix formation in the ArsR-D52N, ΔnikR ΔarsS, and Δfur ΔnikR ΔarsS mutant strains, suggesting that these strains may have an exacerbated stress response that further contributes to biofilm formation. Thus, H. pylori ArsRS has a previously unrecognized role in biofilm formation. IMPORTANCE Despite a paucity of regulatory proteins, adaptation is key to the survival of H. pylori within the stomach. While prior studies have focused on individual

  13. High-Resolution Observations of a Filament showing Activated Barb

    NASA Astrophysics Data System (ADS)

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

    2012-07-01

    Analysis of a filament showing an activated barb using observations from the Dutch Open Telescope (DOT) on 2010 August 20 are presented. The DOT takes Doppler images in Hα, among other wavelengths, in a region about 110 × 110 arcsec^{2} in area, at a cadence of 30~seconds. The offline image restoration technique of speckle reconstruction is applied to obtain diffraction limited images. The filament developed a new barb in 10~minutes, which disappeared within the next 35~minutes. Such a rapid formation and disappearance of a filament barb is unusual, and has not been reported earlier. Line-of-sight velocity maps were constructed from the Doppler images of the target filament. We observe flows in the filament spine towards the barb location prior to its formation, and flows in the barb towards the spine during its disappearance. Photospheric magnetograms from Heliospheric Magnetic Imager on board the Solar Dynamics Observatory, at a cadence of 45~seconds, were used to determine the changes in magnetic flux in the region surrounding the barb location. The variation of magnetic flux in this duration supports the view that barbs are rooted in minor magnetic polarity. Our analysis shows that barbs can be short-lived and formation and disappearance of the barb was associated with cancellation of magnetic flux.

  14. Rab35 regulates cadherin-mediated adherens junction formation and myoblast fusion

    PubMed Central

    Charrasse, Sophie; Comunale, Franck; De Rossi, Sylvain; Echard, Arnaud; Gauthier-Rouvière, Cécile

    2013-01-01

    Cadherins are homophilic cell–cell adhesion molecul