Fabrication of Detector Arrays for the SPT-3G Receiver

NASA Astrophysics Data System (ADS)

Posada, C. M.; Ade, P. A. R.; Ahmed, Z.; Anderson, A. J.; Austermann, J. E.; Avva, J. S.; Thakur, R. Basu; Bender, A. N.; Benson, B. A.; Carlstrom, J. E.; Carter, F. W.; Cecil, T.; Chang, C. L.; Cliche, J. F.; Cukierman, A.; Denison, E. V.; de Haan, T.; Ding, J.; Divan, R.; Dobbs, M. A.; Dutcher, D.; Everett, W.; Foster, A.; Gannon, R. N.; Gilbert, A.; Groh, J. C.; Halverson, N. W.; Harke-Hosemann, A. H.; Harrington, N. L.; Henning, J. W.; Hilton, G. C.; Holzapfel, W. L.; Huang, N.; Irwin, K. D.; Jeong, O. B.; Jonas, M.; Khaire, T.; Kofman, A. M.; Korman, M.; Kubik, D.; Kuhlmann, S.; Kuo, C. L.; Lee, A. T.; Lowitz, A. E.; Meyer, S. S.; Michalik, D.; Miller, C. S.; Montgomery, J.; Nadolski, A.; Natoli, T.; Nguyen, H.; Noble, G. I.; Novosad, V.; Padin, S.; Pan, Z.; Pearson, J.; Rahlin, A.; Ruhl, J. E.; Saunders, L. J.; Sayre, J. T.; Shirley, I.; Shirokoff, E.; Smecher, G.; Sobrin, J. A.; Stan, L.; Stark, A. A.; Story, K. T.; Suzuki, A.; Tang, Q. Y.; Thompson, K. L.; Tucker, C.; Vale, L. R.; Vanderlinde, K.; Vieira, J. D.; Wang, G.; Whitehorn, N.; Yefremenko, V.; Yoon, K. W.; Young, M. R.

2018-05-01

The South Pole Telescope third-generation (SPT-3G) receiver was installed during the austral summer of 2016-2017. It is designed to measure the cosmic microwave background across three frequency bands centered at 95, 150, and 220 GHz. The SPT-3G receiver has ten focal plane modules, each with 269 pixels. Each pixel features a broadband sinuous antenna coupled to a niobium microstrip transmission line. In-line filters define the desired band-passes before the signal is coupled to six bolometers with Ti/Au/Ti/Au transition edge sensors (three bands × two polarizations). In total, the SPT-3G receiver is composed of 16,000 detectors, which are read out using a 68× frequency-domain multiplexing scheme. In this paper, we present the process employed in fabricating the detector arrays.

Behavioural endophenotypes in mice lacking the auxiliary GABAB receptor subunit KCTD16.

PubMed

Cathomas, Flurin; Sigrist, Hannes; Schmid, Luca; Seifritz, Erich; Gassmann, Martin; Bettler, Bernhard; Pryce, Christopher R

2017-01-15

Gamma-aminobutyric acid (GABA) is the major inhibitory neurotransmitter in the brain and is implicated in the pathophysiology of a number of neuropsychiatric disorders. The GABA B receptors are G-protein coupled receptors consisting of principle subunits and auxiliary potassium channel tetramerization domain (KCTD) subunits. The KCTD subunits 8, 12, 12b and 16 are cytosolic proteins that determine the kinetics of the GABA B receptor response. Previously, we demonstrated that Kctd12 null mutant mice (Kctd12 -/- ) exhibit increased auditory fear learning and that Kctd12 +/- mice show altered circadian activity, as well as increased intrinsic excitability in hippocampal pyramidal neurons. KCTD16 has been demonstrated to influence neuronal excitability by regulating GABA B receptor-mediated gating of postsynaptic ion channels. In the present study we investigated for behavioural endophenotypes in Kctd16 -/- and Kctd16 +/- mice. Compared with wild-type (WT) littermates, auditory and contextual fear conditioning were normal in both Kctd16 -/- and Kctd16 +/- mice. When fear memory was tested on the following day, Kctd16 -/- mice exhibited less extinction of auditory fear memory relative to WT and Kctd16 +/- mice, as well as more contextual fear memory relative to WT and, in particular, Kctd16 +/- mice. Relative to WT, both Kctd16 +/- and Kctd16 -/- mice exhibited normal circadian activity. This study adds to the evidence that auxillary KCTD subunits of GABA B receptors contribute to the regulation of behaviours that could constitute endophenotypes for hyper-reactivity to aversive stimuli in neuropsychiatric disorders. Copyright © 2016 Elsevier B.V. All rights reserved.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Marcianò, G.; Huang, D. T., E-mail: d.huang@beatson.gla.ac.uk

The Spt16–SSRP1 heterodimer is a histone chaperone that plays an important role in regulating chromatin assembly. Here, a crystal structure of the N-terminal domain of human Spt16 is presented and it is shown that this domain may contribute to histone binding. The histone chaperone FACT plays an important role in facilitating nucleosome assembly and disassembly during transcription. FACT is a heterodimeric complex consisting of Spt16 and SSRP1. The N-terminal domain of Spt16 resembles an inactive aminopeptidase. How this domain contributes to the histone chaperone activity of FACT remains elusive. Here, the crystal structure of the N-terminal domain (NTD) of humanmore » Spt16 is reported at a resolution of 1.84 Å. The structure adopts an aminopeptidase-like fold similar to those of the Saccharomyces cerevisiae and Schizosaccharomyces pombe Spt16 NTDs. Isothermal titration calorimetry analyses show that human Spt16 NTD binds histones H3/H4 with low-micromolar affinity, suggesting that Spt16 NTD may contribute to histone binding in the FACT complex. Surface-residue conservation and electrostatic analysis reveal a conserved acidic patch that may be involved in histone binding.« less

Subunits of ADA-two-A-containing (ATAC) or Spt-Ada-Gcn5-acetyltrasferase (SAGA) Coactivator Complexes Enhance the Acetyltransferase Activity of GCN5.

PubMed

Riss, Anne; Scheer, Elisabeth; Joint, Mathilde; Trowitzsch, Simon; Berger, Imre; Tora, László

2015-11-27

Histone acetyl transferases (HATs) play a crucial role in eukaryotes by regulating chromatin architecture and locus specific transcription. GCN5 (KAT2A) is a member of the GNAT (Gcn5-related N-acetyltransferase) family of HATs. In metazoans this enzyme is found in two functionally distinct coactivator complexes, SAGA (Spt Ada Gcn5 acetyltransferase) and ATAC (Ada Two A-containing). These two multiprotein complexes comprise complex-specific and shared subunits, which are organized in functional modules. The HAT module of ATAC is composed of GCN5, ADA2a, ADA3, and SGF29, whereas in the SAGA HAT module ADA2b is present instead of ADA2a. To better understand how the activity of human (h) hGCN5 is regulated in the two related, but different, HAT complexes we carried out in vitro HAT assays. We compared the activity of hGCN5 alone with its activity when it was part of purified recombinant hATAC or hSAGA HAT modules or endogenous hATAC or hSAGA complexes using histone tail peptides and full-length histones as substrates. We demonstrated that the subunit environment of the HAT complexes into which GCN5 incorporates determines the enhancement of GCN5 activity. On histone peptides we show that all the tested GCN5-containing complexes acetylate mainly histone H3K14. Our results suggest a stronger influence of ADA2b as compared with ADA2a on the activity of GCN5. However, the lysine acetylation specificity of GCN5 on histone tails or full-length histones was not changed when incorporated in the HAT modules of ATAC or SAGA complexes. Our results thus demonstrate that the catalytic activity of GCN5 is stimulated by subunits of the ADA2a- or ADA2b-containing HAT modules and is further increased by incorporation of the distinct HAT modules in the ATAC or SAGA holo-complexes. © 2015 by The American Society for Biochemistry and Molecular Biology, Inc.

Integrated performance of a frequency domain multiplexing readout in the SPT-3G receiver

NASA Astrophysics Data System (ADS)

Bender, A. N.; Ade, P. A. R.; Anderson, A. J.; Avva, J.; Ahmed, Z.; Arnold, K.; Austermann, J. E.; Basu Thakur, R.; Benson, B. A.; Bleem, L. E.; Byrum, K.; Carlstrom, J. E.; Carter, F. W.; Chang, C. L.; Cho, H. M.; Cliche, J. F.; Crawford, T. M.; Cukierman, A.; Czaplewski, D. A.; Ding, J.; Divan, R.; de Haan, T.; Dobbs, M. A.; Dutcher, D.; Everett, W.; Gilbert, A.; Groh, J. C.; Guyser, R.; Halverson, N. W.; Harke-Hosemann, A.; Harrington, N. L.; Hattori, K.; Henning, J. W.; Hilton, G. C.; Holzapfel, W. L.; Huang, N.; Irwin, K. D.; Jeong, O.; Khaire, T.; Korman, M.; Kubik, D.; Kuo, C. L.; Lee, A. T.; Leitch, E. M.; Lendinez, S.; Meyer, S. S.; Miller, C. S.; Montgomery, J.; Nadolski, A.; Natoli, T.; Nguyen, H.; Novosad, V.; Padin, S.; Pan, Z.; Pearson, J.; Posada, C. M.; Rahlin, A.; Reichardt, C. L.; Ruhl, J. E.; Saliwanchik, B. R.; Sayre, J. T.; Shariff, J. A.; Shirley, Ian; Shirokoff, E.; Smecher, G.; Sobrin, J.; Stan, L.; Stark, A. A.; Story, K.; Suzuki, A.; Tang, Q. Y.; Thompson, K. L.; Tucker, C.; Vanderlinde, K.; Vieira, J. D.; Wang, G.; Whitehorn, N.; Yefremenko, V.; Yoon, K. W.

2016-07-01

The third generation receiver for the South Pole Telescope, SPT-3G, will make extremely deep, arcminuteresolution maps of the temperature and polarization of the cosmic microwave background. The SPT-3G maps will enable studies of the B-mode polarization signature, constraining primordial gravitational waves as well as the effect of massive neutrinos on structure formation in the late universe. The SPT-3G receiver will achieve exceptional sensitivity through a focal plane of 16,000 transition-edge sensor bolometers, an order of magnitude more than the current SPTpol receiver. SPT-3G uses a frequency domain multiplexing (fMux) scheme to read out the focal plane, combining the signals from 64 bolometers onto a single pair of wires. The fMux readout facilitates the large number of detectors in the SPT-3G focal plane by limiting the thermal load due to readout wiring on the 250 millikelvin cryogenic stage. A second advantage of the fMux system is that the operation of each bolometer can be optimized. In addition to these benefits, the fMux readout introduces new challenges into the design and operation of the receiver. The bolometers are operated at a range of frequencies up to 5 MHz, requiring control of stray reactances over a large bandwidth. Additionally, crosstalk between multiplexed detectors will inject large false signals into the data if not adequately mitigated. SPT-3G is scheduled to deploy to the South Pole Telescope in late 2016. Here, we present the pre-deployment performance of the fMux readout system with the SPT-3G focal plane.

Power Electronics Development for the SPT-100 Thruster

NASA Technical Reports Server (NTRS)

Hamley, John A.; Hill, Gerald M.; Sankovic, John M.

1994-01-01

Russian electric propulsion technologies have recently become available on the world market. Of significant interest is the Stationary Plasma Thruster (SPT) which has a significant flight heritage in the former Soviet space program. The SPT has performance levels of up to 1600 seconds of specific impulse at a thrust efficiency of 0.50. Studies have shown that this level of performance is well suited for stationkeeping applications, and the SPT-100, with a 1.35 kW input power level, is presently being evaluated for use on Western commercial satellites. Under a program sponsored by the Innovative Science and Technology Division of the Ballistic Missile Defense Organization, a team of U.S. electric propulsion specialists observed the operation of the SPT-100 in Russia. Under this same program, power electronics were developed to operate the SPT-100 to characterize thruster performance and operation in the U.S. The power electronics consisted of a discharge, cathode heater, and pulse igniter power supplies to operate the thruster with manual flow control. A Russian designed matching network was incorporated in the discharge supply to ensure proper operation with the thruster. The cathode heater power supply and igniter were derived from ongoing development projects. No attempts were made to augment thruster electromagnet current in this effort. The power electronics successfully started and operated the SPT-100 thruster in performance tests at NASA Lewis, with minimal oscillations in the discharge current. The efficiency of the main discharge supply was measured at 0.92, and straightforward modifications were identified which could increase the efficiency to 0.94.

The yeast transcription elongation factor Spt4/5 is a sequence‐specific RNA binding protein

PubMed Central

Blythe, Amanda J.; Yazar‐Klosinski, Berra; Webster, Michael W.; Chen, Eefei; Vandevenne, Marylène; Bendak, Katerina; Mackay, Joel P.; Hartzog, Grant A.

2016-01-01

Abstract The heterodimeric transcription elongation factor Spt4/Spt5 (Spt4/5) tightly associates with RNAPII to regulate both transcriptional elongation and co‐transcriptional pre‐mRNA processing; however, the mechanisms by which Spt4/5 acts are poorly understood. Recent studies of the human and Drosophila Spt4/5 complexes indicate that they can bind nucleic acids in vitro. We demonstrate here that yeast Spt4/5 can bind in a sequence‐specific manner to single stranded RNA containing AAN repeats. Furthermore, we show that the major protein determinants for RNA‐binding are Spt4 together with the NGN domain of Spt5 and that the KOW domains are not required for RNA recognition. These findings attribute a new function to a domain of Spt4/5 that associates directly with RNAPII, making significant steps towards elucidating the mechanism behind transcriptional control by Spt4/5. PMID:27376968

Small Subunits of Serine Palmitoyltransferase (ssSPTs) and Their Physiological Roles

DTIC Science & Technology

2014-02-12

showing that organisms also have unique sphingoid base chain lengths. In insects, such as Drosophila melanogaster , the predominant chain lengths of the ... Drosophila melanogaster mutant defective in male meiotic cytokinesis (‘Ghiberti’) has a mutation in a gene with low homology to the ssSPT subunits of...INTRODUCTION: Sphingolipid metabolism in Drosophila melanogaster (fly) is an active area of research. It is a good model system to study the roles of

Spt6 Association with RNA Polymerase II Directs mRNA Turnover During Transcription.

PubMed

Dronamraju, Raghuvar; Hepperla, Austin J; Shibata, Yoichiro; Adams, Alexander T; Magnuson, Terry; Davis, Ian J; Strahl, Brian D

2018-06-21

Spt6 is an essential histone chaperone that mediates nucleosome reassembly during gene transcription. Spt6 also associates with RNA polymerase II (RNAPII) via a tandem Src2 homology domain. However, the significance of Spt6-RNAPII interaction is not well understood. Here, we show that Spt6 recruitment to genes and the nucleosome reassembly functions of Spt6 can still occur in the absence of its association with RNAPII. Surprisingly, we found that Spt6-RNAPII association is required for efficient recruitment of the Ccr4-Not de-adenylation complex to transcribed genes for essential degradation of a range of mRNAs, including mRNAs required for cell-cycle progression. These findings reveal an unexpected control mechanism for mRNA turnover during transcription facilitated by a histone chaperone. Copyright © 2018 Elsevier Inc. All rights reserved.

VizieR Online Data Catalog: SPT-GMOS spectroscopy of gal. in massive clusters (Bayliss+, 2017)

NASA Astrophysics Data System (ADS)

Bayliss, M. B.; Zengo, K.; Ruel, J.; Benson, B. A.; Bleem, L. E.; Bocquet, S.; Bulbul, E.; Brodwin, M.; Capasso, R.; Chiu, I.-N.; McDonald, M.; Rapetti, D.; Saro, A.; Stalder, B.; Stark, A. A.; Strazzullo, V.; Stubbs, C. W.; Zenteno, A.

2017-10-01

The majority of the data set used in this analysis comes from the SPT-GMOS spectroscopic survey (Bayliss+ 2016, J/ApJS/227/3), which consists of spectroscopic follow-up of 62 galaxy clusters from the SPT-SZ survey. The full SPT-GMOS sample includes 2243 galaxy spectra, 1579 of which are cluster member galaxies. In addition to previously published galaxy spectroscopy, we also present the first publication of new spectroscopy in the fields of three SPT galaxy clusters. We observed SPT-CLJ0000-5748, SPT-CLJ0516-5430, and SPT-CLJ2337-5942 with the Inamori Magellan Areal Camera and Spectrograph (IMACS) mounted on the Magellan-I (Baade) telescope at Las Campanas Observatory on the nights of 14-15 September 2012. (2 data files).

VizieR Online Data Catalog: Full spectroscopic data release of the SPT-GMOS (Bayliss+, 2016)

NASA Astrophysics Data System (ADS)

Bayliss, M. B.; Ruel, J.; Stubbs, C. W.; Allen, S. W.; Applegate, D. E.; Ashby, M. L. N.; Bautz, M.; Benson, B. A.; Bleem, L. E.; Bocquet, S.; Brodwin, M.; Capasso, R.; Carlstrom, J. E.; Chang, C. L.; Chiu, I.; Cho, H.-M.; Clocchiatti, A.; Crawford, T. M.; Crites, A. T.; de Haan, T.; Desai, S.; Dietrich, J. P.; Dobbs, M. A.; Doucouliagos, A. N.; Foley, R. J.; Forman, W. R.; Garmire, G. P.; George, E. M.; Gladders, M. D.; Gonzalez, A. H.; Gupta, N.; Halverson, N. W.; Hlavacek-Larrondo, J.; Hoekstra, H.; Holder, G. P.; Holzapfel, W. L.; Hou, Z.; Hrubes, J. D.; Huang, N.; Jones, C.; Keisler, R.; Knox, L.; Lee, A. T.; Leitch, E. M.; von der Linden, A.; Luong-van, D.; Mantz, A.; Marrone, D. P.; McDonald, M.; McMahon, J. J.; Meyer, S. S.; Mocanu, L. M.; Mohr, J. J.; Murray, S. S.; Padin, S.; Pryke, C.; Rapetti, D.; Reichardt, C. L.; Rest, A.; Ruhl, J. E.; Saliwanchik, B. R.; Saro, A.; Sayre, J. T.; Schaffer, K. K.; Schrabback, T.; Shirokoff, E.; Song, J.; Spieler, H. G.; Stalder, B.; Stanford, S. A.; Staniszewski, Z.; Stark, A. A.; Story, K. T.; Vanderlinde, K.; Vieira, J. D.; Vikhlinin, A.; Williamson, R.; Zenteno, A.

2016-11-01

The data presented in this paper follow the same observational design described by Ruel et al. (2014ApJ...792...45R). The final Gemini-S observing allocation for SPT-GMOS concluded at the end of the 2015B semester. Over the course of the entire survey we observed 121 individual spectroscopic masks targeting 62 SPT-SZ galaxy clusters (see table 1). All final data products from SPT-GMOS are publicly released via the Harvard Dataverse Network (http://dataverse.harvard.edu/dataverse/SPT_Clusters). The galaxy clusters observed in the SPT-GMOS are all drawn from the SPT-SZ survey, completed in 2011 November. The full SPT-SZ survey covered approximately 2500deg2 of the southern sky at 95, 150, and 220GHz with an angular resolution of ~1'. (4 data files).

Faithful chromosome transmission requires Spt4p, a putative regulator of chromatin structure in Saccharomyces cerevisiae.

PubMed Central

Basrai, M A; Kingsbury, J; Koshland, D; Spencer, F; Hieter, P

1996-01-01

A chromosome transmission fidelity (ctf) mutant, s138, of Saccharomyces cerevisiae was identified by its centromere (CEN) transcriptional readthrough phenotype, suggesting perturbed kinetochore integrity in vivo. The gene complementing the s138 mutation was found to be identical to the S. cerevisiae SPT4 gene. The s138 mutation is a missense mutation in the second of four conserved cysteine residues positioned similarly to those of zinc finger proteins, and we henceforth refer to the mutation of spt4-138. Both spt4-138 and spt4 delta strains missegregate a chromosome fragment at the permissive temperature, are temperature sensitive for growth at 37 degrees C, and upon a shift to the nonpermissive temperature show an accumulation of large budded cells, each with a nucleus. Previous studies suggest that Spt4p functions in a complex with Spt5p and Spt6p, and we determined that spt6-140 also causes missegregation of a chromosome fragment. Double mutants carrying spt4 delta 2::HIS3 and kinetochore mutation ndc10-42 or ctf13-30 show a synthetic conditional phenotype. Both spt4-138 and spt4 delta strains exhibit synergistic chromosome instability in combination with CEN DNA mutations and show in vitro defects in microtubule binding to minichromosomes. These results indicate that Spt4p plays a role in chromosome segregation. The results of in vivo genetic interactions with mutations in kinetochore proteins and CEN DNA and of in vitro biochemical assays suggest that Spt4p is important for kinetochore function. PMID:8649393

Immunogenicity and protective efficacy of a Salmonella Enteritidis sptP mutant as a live attenuated vaccine candidate.

PubMed

Lin, Zhijie; Tang, Peipei; Jiao, Yang; Kang, Xilong; Li, Qiuchun; Xu, Xiulong; Sun, Jun; Pan, Zhiming; Jiao, Xinan

2017-06-24

Salmonella enterica serovar Enteritidis (S. Enteritidis) is a highly adaptive pathogen in both humans and animals. As a Salmonella Type III secretion system (T3SS) effector, Salmonella protein tyrosine phosphatase (SptP) is critical for virulence in this genus. To investigate the feasibility of using C50336ΔsptP as a live attenuated oral vaccine in mice, we generated the sptP gene deletion mutant C50336ΔsptP in S. Enteritidis strain C50336 by λ-Red mediated recombination and evaluated the protective ability of the S. Enteritidis sptP mutant strain C50336ΔsptP against mice salmonellosis. We found that C50336ΔsptP was a highly immunogenic, effective, and safe vaccine in mice. Compared to wild-type C50336, C50336ΔsptP showed reduced virulence as confirmed by the 50% lethal dose (LD 50 ) in orally infected mice. C50336ΔsptP also showed decreased bacterial colonization both in vivo and in vitro. Immunization with C50336ΔsptP had no significant effect on body weight and did not result in obvious clinical symptoms relative to control animals treated with phosphate-buffered saline (PBS), but induced humoral and cellular immune responses at 12 and 26 days post inoculation. Immunization with 1 × 10 8 colony-forming units (CFU) C50336ΔsptP per mouse provided 100% protection against subsequent challenge with the wild-type C50336 strain, and immunized mice showed mild and temporary clinical symptoms as compared to those of control group. These results demonstrate that C50336ΔsptP can be a live attenuated oral vaccine for salmonellosis.

Supportive periodontal therapy (SPT) for maintaining the dentition in adults treated for periodontitis.

PubMed

Manresa, Carolina; Sanz-Miralles, Elena C; Twigg, Joshua; Bravo, Manuel

2018-01-01

Periodontitis is a bacterially-induced, chronic inflammatory disease that destroys the connective tissues and bone that support teeth. Active periodontal treatment aims to reduce the inflammatory response, primarily through eradication of bacterial deposits. Following completion of treatment and arrest of inflammation, supportive periodontal therapy (SPT) is employed to reduce the probability of re-infection and progression of the disease; to maintain teeth without pain, excessive mobility or persistent infection in the long term, and to prevent related oral diseases.According to the American Academy of Periodontology, SPT should include all components of a typical dental recall examination, and importantly should also include periodontal re-evaluation and risk assessment, supragingival and subgingival removal of bacterial plaque and calculus, and re-treatment of any sites showing recurrent or persistent disease. While the first four points might be expected to form part of the routine examination appointment for periodontally healthy patients, the inclusion of thorough periodontal evaluation, risk assessment and subsequent treatment - normally including mechanical debridement of any plaque or calculus deposits - differentiates SPT from routine care.Success of SPT has been reported in a number of long-term, retrospective studies. This review aimed to assess the evidence available from randomised controlled trials (RCTs). To determine the effects of supportive periodontal therapy (SPT) in the maintenance of the dentition of adults treated for periodontitis. Cochrane Oral Health's Information Specialist searched the following databases: Cochrane Oral Health's Trials Register (to 8 May 2017), the Cochrane Central Register of Controlled Trials (CENTRAL) (the Cochrane Library, 2017, Issue 5), MEDLINE Ovid (1946 to 8 May 2017), and Embase Ovid (1980 to 8 May 2017). The US National Institutes of Health Trials Registry (ClinicalTrials.gov) and the World Health Organization

Design and Assembly of SPT-3G Cold Readout Hardware

NASA Astrophysics Data System (ADS)

Avva, J. S.; Ade, P. A. R.; Ahmed, Z.; Anderson, A. J.; Austermann, J. E.; Thakur, R. Basu; Barron, D.; Bender, A. N.; Benson, B. A.; Carlstrom, J. E.; Carter, F. W.; Cecil, T.; Chang, C. L.; Cliche, J. F.; Cukierman, A.; Denison, E. V.; de Haan, T.; Ding, J.; Dobbs, M. A.; Dutcher, D.; Elleflot, T.; Everett, W.; Foster, A.; Gannon, R. N.; Gilbert, A.; Groh, J. C.; Halverson, N. W.; Harke-Hosemann, A. H.; Harrington, N. L.; Hasegawa, M.; Hattori, K.; Henning, J. W.; Hilton, G. C.; Holzapfel, W. L.; Hori, Y.; Huang, N.; Irwin, K. D.; Jeong, O. B.; Jonas, M.; Khaire, T.; Kofman, A. M.; Korman, M.; Kubik, D.; Kuhlmann, S.; Kuo, C. L.; Lee, A. T.; Lowitz, A. E.; Meyer, S. S.; Montgomery, J.; Nadolski, A.; Natoli, T.; Nguyen, H.; Nishino, H.; Noble, G. I.; Novosad, V.; Padin, S.; Pan, Z.; Pearson, J.; Posada, C. M.; Rahlin, A.; Rotermund, K.; Ruhl, J. E.; Saunders, L. J.; Sayre, J. T.; Shirley, I.; Shirokoff, E.; Smecher, G.; Sobrin, J. A.; Stark, A. A.; Story, K. T.; Suzuki, A.; Tang, Q. Y.; Thompson, K. L.; Tucker, C.; Vale, L. R.; Vanderlinde, K.; Vieira, J. D.; Wang, G.; Whitehorn, N.; Yefremenko, V.; Yoon, K. W.; Young, M. R.

2018-05-01

The third-generation upgrade to the receiver on the South Pole Telescope, SPT-3G, was installed at the South Pole during the 2016-2017 austral summer to measure the polarization of the cosmic microwave background. Increasing the number of detectors by a factor of 10 to ˜ 16,000 required the multiplexing factor to increase to 68 and the bandwidth of the frequency-division readout electronics to span 1.6-5.2 MHz. This increase necessitates low-thermal conductance, low-inductance cryogenic wiring. Our cold readout system consists of planar thin-film aluminum inductive-capacitive resonators, wired in series with the detectors, summed together, and connected to 4K SQUIDs by 10-μm -thick niobium-titanium (NbTi) broadside-coupled striplines. Here, we present an overview of the cold readout electronics for SPT-3G, including assembly details and characterization of electrical and thermal properties of the system. We report, for the NbTi striplines, values of R ≤ 10^{-4} Ω , L = 21 ± 1 nH , and C = 1.47± .02 nF . Additionally, the striplines' thermal conductivity is described by kA = 6.0± 0.3 T^{0.92 ± 0.04} μW mm K^{-1} . Finally, we provide projections for cross talk induced by parasitic impedances from the stripline and find that the median value of percentage cross talk from leakage current is 0.22 and 0.09% from wiring impedance.

The Arabidopsis mediator complex subunits MED16, MED14, and MED2 regulate mediator and RNA polymerase II recruitment to CBF-responsive cold-regulated genes.

PubMed

Hemsley, Piers A; Hurst, Charlotte H; Kaliyadasa, Ewon; Lamb, Rebecca; Knight, Marc R; De Cothi, Elizabeth A; Steele, John F; Knight, Heather

2014-01-01

The Mediator16 (MED16; formerly termed SENSITIVE TO FREEZING6 [SFR6]) subunit of the plant Mediator transcriptional coactivator complex regulates cold-responsive gene expression in Arabidopsis thaliana, acting downstream of the C-repeat binding factor (CBF) transcription factors to recruit the core Mediator complex to cold-regulated genes. Here, we use loss-of-function mutants to show that RNA polymerase II recruitment to CBF-responsive cold-regulated genes requires MED16, MED2, and MED14 subunits. Transcription of genes known to be regulated via CBFs binding to the C-repeat motif/drought-responsive element promoter motif requires all three Mediator subunits, as does cold acclimation-induced freezing tolerance. In addition, these three subunits are required for low temperature-induced expression of some other, but not all, cold-responsive genes, including genes that are not known targets of CBFs. Genes inducible by darkness also required MED16 but required a different combination of Mediator subunits for their expression than the genes induced by cold. Together, our data illustrate that plants control transcription of specific genes through the action of subsets of Mediator subunits; the specific combination defined by the nature of the stimulus but also by the identity of the gene induced.

Optical spectroscopy and velocity dispersions of galaxy clusters from the SPT-SZ survey

DOE Office of Scientific and Technical Information (OSTI.GOV)

Ruel, J.; Bayliss, M.; Bazin, G.

2014-09-01

We present optical spectroscopy of galaxies in clusters detected through the Sunyaev-Zel'dovich (SZ) effect with the South Pole Telescope (SPT). We report our own measurements of 61 spectroscopic cluster redshifts, and 48 velocity dispersions each calculated with more than 15 member galaxies. This catalog also includes 19 dispersions of SPT-observed clusters previously reported in the literature. The majority of the clusters in this paper are SPT-discovered; of these, most have been previously reported in other SPT cluster catalogs, and five are reported here as SPT discoveries for the first time. By performing a resampling analysis of galaxy velocities, we findmore » that unbiased velocity dispersions can be obtained from a relatively small number of member galaxies (≲ 30), but with increased systematic scatter. We use this analysis to determine statistical confidence intervals that include the effect of membership selection. We fit scaling relations between the observed cluster velocity dispersions and mass estimates from SZ and X-ray observables. In both cases, the results are consistent with the scaling relation between velocity dispersion and mass expected from dark-matter simulations. We measure a ∼30% log-normal scatter in dispersion at fixed mass, and a ∼10% offset in the normalization of the dispersion-mass relation when compared to the expectation from simulations, which is within the expected level of systematic uncertainty.« less

SPT-3G: a next-generation cosmic microwave background polarization experiment on the South Pole telescope

NASA Astrophysics Data System (ADS)

Benson, B. A.; Ade, P. A. R.; Ahmed, Z.; Allen, S. W.; Arnold, K.; Austermann, J. E.; Bender, A. N.; Bleem, L. E.; Carlstrom, J. E.; Chang, C. L.; Cho, H. M.; Cliche, J. F.; Crawford, T. M.; Cukierman, A.; de Haan, T.; Dobbs, M. A.; Dutcher, D.; Everett, W.; Gilbert, A.; Halverson, N. W.; Hanson, D.; Harrington, N. L.; Hattori, K.; Henning, J. W.; Hilton, G. C.; Holder, G. P.; Holzapfel, W. L.; Irwin, K. D.; Keisler, R.; Knox, L.; Kubik, D.; Kuo, C. L.; Lee, A. T.; Leitch, E. M.; Li, D.; McDonald, M.; Meyer, S. S.; Montgomery, J.; Myers, M.; Natoli, T.; Nguyen, H.; Novosad, V.; Padin, S.; Pan, Z.; Pearson, J.; Reichardt, C.; Ruhl, J. E.; Saliwanchik, B. R.; Simard, G.; Smecher, G.; Sayre, J. T.; Shirokoff, E.; Stark, A. A.; Story, K.; Suzuki, A.; Thompson, K. L.; Tucker, C.; Vanderlinde, K.; Vieira, J. D.; Vikhlinin, A.; Wang, G.; Yefremenko, V.; Yoon, K. W.

2014-07-01

We describe the design of a new polarization sensitive receiver, spt-3g, for the 10-meter South Pole Telescope (spt). The spt-3g receiver will deliver a factor of ~20 improvement in mapping speed over the current receiver, spt-pol. The sensitivity of the spt-3g receiver will enable the advance from statistical detection of B-mode polarization anisotropy power to high signal-to-noise measurements of the individual modes, i.e., maps. This will lead to precise (~0.06 eV) constraints on the sum of neutrino masses with the potential to directly address the neutrino mass hierarchy. It will allow a separation of the lensing and inflationary B-mode power spectra, improving constraints on the amplitude and shape of the primordial signal, either through spt-3g data alone or in combination with bicep2/keck, which is observing the same area of sky. The measurement of small-scale temperature anisotropy will provide new constraints on the epoch of reionization. Additional science from the spt-3g survey will be significantly enhanced by the synergy with the ongoing optical Dark Energy Survey (des), including: a 1% constraint on the bias of optical tracers of large-scale structure, a measurement of the differential Doppler signal from pairs of galaxy clusters that will test General Relativity on ~200Mpc scales, and improved cosmological constraints from the abundance of clusters of galaxies

Ubiquitin fusion constructs allow the expression and purification of multi-KOW domain complexes of the Saccharomyces cerevisiae transcription elongation factor Spt4/5.

PubMed

Blythe, Amanda; Gunasekara, Sanjika; Walshe, James; Mackay, Joel P; Hartzog, Grant A; Vrielink, Alice

2014-08-01

Spt4/5 is a hetero-dimeric transcription elongation factor that can both inhibit and promote transcription elongation by RNA polymerase II (RNAPII). However, Spt4/5's mechanism of action remains elusive. Spt5 is an essential protein and the only universally-conserved RNAP-associated transcription elongation factor. The protein contains multiple Kyrpides, Ouzounis and Woese (KOW) domains. These domains, in other proteins, are thought to bind RNA although there is little direct evidence in the literature to support such a function in Spt5. This could be due, at least in part, to difficulties in expressing and purifying recombinant Spt5. When expressed in Escherichia coli (E. coli), Spt5 is innately insoluble. Here we report a new approach for the successful expression and purification of milligram quantities of three different multi-KOW domain complexes of Saccharomyces cerevisiae Spt4/5 for use in future functional studies. Using the E. coli strain Rosetta2 (DE3) we have developed strategies for co-expression of Spt4 and multi-KOW domain Spt5 complexes from the bi-cistronic pET-Duet vector. In a second strategy, Spt4/5 was expressed via co-transformation of Spt4 in the vector pET-M11 with Spt5 ubiquitin fusion constructs in the vector pHUE. We characterized the multi-KOW domain Spt4/5 complexes by Western blot, limited proteolysis, circular dichroism, SDS-PAGE and size exclusion chromatography-multiangle light scattering and found that the proteins are folded with a Spt4:Spt5 hetero-dimeric stoichiometry of 1:1. These expression constructs encompass a larger region of Spt5 than has previously been reported, and will provide the opportunity to elucidate the biological function of the multi-KOW containing Spt5. Copyright © 2014 Elsevier Inc. All rights reserved.

Preventative effect of OMZ-SPT on lipopolysaccharide-induced acute lung injury and inflammation via nuclear factor-kappa B signaling in mice

DOE Office of Scientific and Technical Information (OSTI.GOV)

Wang, Ting; Department of Respiratory Medicine, Children's Hospital of Chongqing Medical University, Chongqing, 400014; Hou, Wanru

Acute lung injury (ALI) is an early pathophysiologic change in acute respiratory distress syndrome and its management can be challenging. Omalizumab (Xolair™) is a recombinant DNA-derived, humanized antibody. OMZ-SPT is a polypeptide on the heavy chain of omalizumab monoclonal antibody. Here, we found that intramuscular administration of OMZ-SPT significantly improved survival and attenuated lung inflammation in female C57BL/6 mice suffering from lipopolysaccharide (LPS)-induced ALI. We also demonstrated that OMZ-SPT can inhibit expression of the inflammatory cytokines tumor necrosis factor-α, interleukin-1β and interleukin-6 by ELISA in mice suffering from LPS-induced ALI and a mouse macrophage line (RAW264.7 cells). In addition, we showedmore » that OMZ-SPT inhibited LPS-induced activation of nuclear factor-kappa B (NF-κB) signaling and total expression of NF-κB by western blotting. These data suggest that OMZ-SPT could be a novel therapeutic choice for ALI. - Highlights: • OMZ-SPT is a polypeptide on the heavy chain of omalizumab monoclonal antibody. • Omalizumab (Xolair™) have anti-inflammatory effects. • OMZ-SPT can inhibit inflammatory responses and lung injury in LPS-induced ALI mice. • Protective effect of OMZ-SPT on ALI is due to inhibition of NF-κB signaling. • OMZ-SPT could be a novel therapeutic choice for ALI.« less

Component-resolved diagnosis of plant food allergy by SPT.

PubMed

Asero, R; Jimeno, L; Barber, D

2008-12-01

Fruits and vegetables may contain both labile and stable allergens. The former induce only OAS, whereas stable allergens may induce systemic reactions. Component-resolved diagnosis (CRD) of allergy to plant foods is therefore essential for the clinical management of allergic patients. 80 adults allergic to plant foods underwent SPT with purified natural date palm profilin (Pho d 2), purified Mal d 1, a peach extract containing uniquely LTP, and with a kiwi extract containing uniquely stable allergens. 58 (72%) patients were monosensitized: 24 to Mal d 1, 24 to profilin, 7 to LTP, and 3 to kiwi. 22 patients were multi-sensitised: 14 to Mal d 1 and profilin, 2 to Mal d 1 and kiwi, 1 to LTP and profilin, 3 to LTP and Mal d 1, and 2 to LTP, Mal d 1 and profilin. Mal d 1 and LTP sensitisation were associated with apple and peach allergy, respectively, whereas profilin sensitisation was associated with allergy to melon, watermelon, banana, tomato and citrus fruits. 18/21 kiwi-allergic patients were sensitised to one of the cross-reacting allergens, but 2/18 reacted to kiwi-specific allergens as well. In patients with allergy to plant-derived foods CRD can be performed by SPT with purified allergen proteins. In the future, the availability of a larger number of purified natural or recombinant allergens for SPT will represent a simple means to classify food-allergic patients properly on the first visit.

Fabrication of large dual-polarized multichroic TES bolometer arrays for CMB measurements with the SPT-3G camera

DOE Office of Scientific and Technical Information (OSTI.GOV)

Posada, C. M.; Ade, P. A. R.; Ahmed, Z.

2015-08-11

This work presents the procedures used by Argonne National Laboratory to fabricate large arrays of multichroic transition-edge sensor (TES) bolometers for cosmic microwave background (CMB) measurements. These detectors will be assembled into the focal plane for the SPT-3G camera, the third generation CMB camera to be installed in the South Pole Telescope. The complete SPT-3G camera will have approximately 2690 pixels, for a total of 16,140 TES bolometric detectors. Each pixel is comprised of a broad-band sinuous antenna coupled to a Nb microstrip line. In-line filters are used to define the different band-passes before the millimeter-wavelength signal is fed tomore » the respective Ti/Au TES bolometers. There are six TES bolometer detectors per pixel, which allow for measurements of three band-passes (95 GHz, 150 GHz and 220 GHz) and two polarizations. The steps involved in the monolithic fabrication of these detector arrays are presented here in detail. Patterns are defined using a combination of stepper and contact lithography. The misalignment between layers is kept below 200 nm. The overall fabrication involves a total of 16 processes, including reactive and magnetron sputtering, reactive ion etching, inductively coupled plasma etching and chemical etching.« less

The Arabidopsis Mediator Complex Subunits MED16, MED14, and MED2 Regulate Mediator and RNA Polymerase II Recruitment to CBF-Responsive Cold-Regulated Genes[C][W][OPEN

PubMed Central

Hemsley, Piers A.; Hurst, Charlotte H.; Kaliyadasa, Ewon; Lamb, Rebecca; Knight, Marc R.; De Cothi, Elizabeth A.; Steele, John F.; Knight, Heather

2014-01-01

The Mediator16 (MED16; formerly termed SENSITIVE TO FREEZING6 [SFR6]) subunit of the plant Mediator transcriptional coactivator complex regulates cold-responsive gene expression in Arabidopsis thaliana, acting downstream of the C-repeat binding factor (CBF) transcription factors to recruit the core Mediator complex to cold-regulated genes. Here, we use loss-of-function mutants to show that RNA polymerase II recruitment to CBF-responsive cold-regulated genes requires MED16, MED2, and MED14 subunits. Transcription of genes known to be regulated via CBFs binding to the C-repeat motif/drought-responsive element promoter motif requires all three Mediator subunits, as does cold acclimation–induced freezing tolerance. In addition, these three subunits are required for low temperature–induced expression of some other, but not all, cold-responsive genes, including genes that are not known targets of CBFs. Genes inducible by darkness also required MED16 but required a different combination of Mediator subunits for their expression than the genes induced by cold. Together, our data illustrate that plants control transcription of specific genes through the action of subsets of Mediator subunits; the specific combination defined by the nature of the stimulus but also by the identity of the gene induced. PMID:24415770

Low-Power Operation and Plasma Characterization of a Qualification Model SPT-140 Hall Thruster for NASA Science Missions

NASA Technical Reports Server (NTRS)

Garner, Charles E.; Jorns, Benjamin A.; van Derventer, Steven; Hofer, Richard R.; Rickard, Ryan; Liang, Raymond; Delgado, Jorge

2015-01-01

Hall thruster systems based on commercial product lines can potentially lead to lower cost electric propulsion (EP) systems for deep space science missions. A 4.5-kW SPT-140 Hall thruster presently under qualification testing by SSL leverages the substantial heritage of the SPT-100 being flown on Russian and US commercial satellites. The Jet Propulsion Laboratory is exploring the use of commercial EP systems, including the SPT-140, for deep space science missions, and initiated a program to evaluate the SPT-140 in the areas of low power operation and thruster operating life. A qualification model SPT-140 designated QM002 was evaluated for operation and plasma properties along channel centerline, from 4.5 kW to 0.8 kW. Additional testing was performed on a development model SPT-140 designated DM4 to evaluate operation with a Moog proportional flow control valve (PFCV). The PFCV was commanded by an SSL engineering model PPU-140 Power Processing Unit (PPU). Performance measurements on QM002 at 0.8 kW discharge power were 50 mN of thrust at a total specific impulse of 1250 s, a total thruster efficiency of 0.38, and discharge current oscillations of under 3% of the mean current. Steady-state operation at 0.8 kW was demonstrated during a 27 h firing. The SPT-140 DM4 was operated in closed-loop control of the discharge current with the PFCV and PPU over discharge power levels of 0.8-4.5 kW. QM002 and DM4 test data indicate that the SPT-140 design is a viable candidate for NASA missions requiring power throttling down to low thruster input power.

Optical–SZE scaling relations for DES optically selected clusters within the SPT-SZ Survey

DOE PAGES

Saro, A.; Bocquet, S.; Mohr, J.; ...

2017-03-15

We study the Sunyaev-Zel'dovich effect (SZE) signature in South Pole Telescope (SPT) data for an ensemble of 719 optically identified galaxy clusters selected from 124.6 degmore » $^2$ of the Dark Energy Survey (DES) science verification data, detecting a stacked SZE signal down to richness $$\\lambda\\sim20$$. The SZE signature is measured using matched-filtered maps of the 2500 deg$^2$ SPT-SZ survey at the positions of the DES clusters, and the degeneracy between SZE observable and matched-filter size is broken by adopting as priors SZE and optical mass-observable relations that are either calibrated using SPT selected clusters or through the Arnaud et al. (2010, A10) X-ray analysis. We measure the SPT signal to noise $$\\zeta$$-$$\\lambda$$, relation and two integrated Compton-$y$ $$Y_\\textrm{500}$$-$$\\lambda$$ relations for the DES-selected clusters and compare these to model expectations accounting for the SZE-optical center offset distribution. For clusters with $$\\lambda > 80$$, the two SPT calibrated scaling relations are consistent with the measurements, while for the A10-calibrated relation the measured SZE signal is smaller by a factor of $$0.61 \\pm 0.12$$ compared to the prediction. For clusters at $$20 < \\lambda < 80$$, the measured SZE signal is smaller by a factor of $$\\sim$$0.20-0.80 (between 2.3 and 10~$$\\sigma$$ significance) compared to the prediction, with the SPT calibrated scaling relations and larger $$\\lambda$$ clusters showing generally better agreement. We quantify the required corrections to achieve consistency, showing that there is a richness dependent bias that can be explained by some combination of contamination of the observables and biases in the estimated masses. We discuss possible physical effects, as contamination from line-of-sight projections or from point sources, larger offsets in the SZE-optical centering or larger scatter in the $$\\lambda$$-mass relation at lower richnesses.« less

Optical–SZE scaling relations for DES optically selected clusters within the SPT-SZ Survey

DOE Office of Scientific and Technical Information (OSTI.GOV)

Saro, A.; Bocquet, S.; Mohr, J.

We study the Sunyaev-Zel'dovich effect (SZE) signature in South Pole Telescope (SPT) data for an ensemble of 719 optically identified galaxy clusters selected from 124.6 degmore » $^2$ of the Dark Energy Survey (DES) science verification data, detecting a stacked SZE signal down to richness $$\\lambda\\sim20$$. The SZE signature is measured using matched-filtered maps of the 2500 deg$^2$ SPT-SZ survey at the positions of the DES clusters, and the degeneracy between SZE observable and matched-filter size is broken by adopting as priors SZE and optical mass-observable relations that are either calibrated using SPT selected clusters or through the Arnaud et al. (2010, A10) X-ray analysis. We measure the SPT signal to noise $$\\zeta$$-$$\\lambda$$, relation and two integrated Compton-$y$ $$Y_\\textrm{500}$$-$$\\lambda$$ relations for the DES-selected clusters and compare these to model expectations accounting for the SZE-optical center offset distribution. For clusters with $$\\lambda > 80$$, the two SPT calibrated scaling relations are consistent with the measurements, while for the A10-calibrated relation the measured SZE signal is smaller by a factor of $$0.61 \\pm 0.12$$ compared to the prediction. For clusters at $$20 < \\lambda < 80$$, the measured SZE signal is smaller by a factor of $$\\sim$$0.20-0.80 (between 2.3 and 10~$$\\sigma$$ significance) compared to the prediction, with the SPT calibrated scaling relations and larger $$\\lambda$$ clusters showing generally better agreement. We quantify the required corrections to achieve consistency, showing that there is a richness dependent bias that can be explained by some combination of contamination of the observables and biases in the estimated masses. We discuss possible physical effects, as contamination from line-of-sight projections or from point sources, larger offsets in the SZE-optical centering or larger scatter in the $$\\lambda$$-mass relation at lower richnesses.« less

The Mini-SPT (Space Particle Telescope) for dual use: Precision flux measurement of low energy proton electron and heavy ion with tracking capability and A compact, low-cost realtime local radiation hazard/alarm detector to be used on board a satellite

NASA Astrophysics Data System (ADS)

Alpat, Behcet; Ergin, Tulun; Kalemci, Emrah

2016-07-01

The Mini-SPT project is the first, and most important, step towards the ambitious goal of creating a low-cost, compact, radiation hardened and high performance space particle telescope that can be mounted, in the near future, as standard particle detector on any satellite. Mini-SPT will be capable of providing high quality physics data on local space environment. In particular high precision flux measurement and tracking of low energy protons and electrons on different orbits with same instrumentation is of paramount importance for studies as geomagnetically trapped fluxes and space weather dynamics, dark matter search, low energy proton anisotropy and its effects on ICs as well as the solar protons studies. In addition, it will provide real-time "differentiable warnings" about the local space radiation hazard to other electronics systems on board the hosting satellite, including different criticality levels and alarm signals to activate mitigation techniques whenever this is strictly necessary to protect them from temporary/permanent failures. A real-time warning system will help satellite subsystems to save significant amount of power and memory with respect to other conventional techniques where the "mitigation" solutions are required to be active during entire mission life. The Mini-SPT will combine the use of technologies developed in cutting-edge high energy physics experiments (including technology from CMS experiments at CERN) and the development of new charged particle detecting systems for their use for the first time in space. The Mini-SPT essential objective is, by using for the first time in space SIPMs (Silicon Photomultipliers) technology for TOF and energy measurements, the production of high quality data with a good time, position and energy resolutions. The mini-SPT will consists of three main sub-units: a- A tracking and dE/dX measuring sub-detector which will be based on silicon pixel detectors (SPD) coupled to the rad-hard chip ROC-DIG (Read

A measurement of the cosmic microwave background damping tail from the 2500-square-degree SPT-SZ survey

DOE Office of Scientific and Technical Information (OSTI.GOV)

Story, K. T.; Keisler, R.; Benson, B. A.

2013-12-10

We present a measurement of the cosmic microwave background (CMB) temperature power spectrum using data from the recently completed South Pole Telescope Sunyaev-Zel'dovich (SPT-SZ) survey. This measurement is made from observations of 2540 deg{sup 2} of sky with arcminute resolution at 150 GHz, and improves upon previous measurements using the SPT by tripling the sky area. We report CMB temperature anisotropy power over the multipole range 650 < ℓ < 3000. We fit the SPT bandpowers, combined with the 7 yr Wilkinson Microwave Anisotropy Probe (WMAP7) data, with a six-parameter ΛCDM cosmological model and find that the two datasets aremore » consistent and well fit by the model. Adding SPT measurements significantly improves ΛCDM parameter constraints; in particular, the constraint on θ {sub s} tightens by a factor of 2.7. The impact of gravitational lensing is detected at 8.1σ, the most significant detection to date. This sensitivity of the SPT+WMAP7 data to lensing by large-scale structure at low redshifts allows us to constrain the mean curvature of the observable universe with CMB data alone to be Ω{sub k}=−0.003{sub −0.018}{sup +0.014}. Using the SPT+WMAP7 data, we measure the spectral index of scalar fluctuations to be n{sub s} = 0.9623 ± 0.0097 in the ΛCDM model, a 3.9σ preference for a scale-dependent spectrum with n{sub s} < 1. The SPT measurement of the CMB damping tail helps break the degeneracy that exists between the tensor-to-scalar ratio r and n{sub s} in large-scale CMB measurements, leading to an upper limit of r < 0.18 (95% C.L.) in the ΛCDM+r model. Adding low-redshift measurements of the Hubble constant (H {sub 0}) and the baryon acoustic oscillation (BAO) feature to the SPT+WMAP7 data leads to further improvements. The combination of SPT+WMAP7+H {sub 0}+BAO constrains n{sub s} = 0.9538 ± 0.0081 in the ΛCDM model, a 5.7σ detection of n{sub s} < 1, and places an upper limit of r < 0.11 (95% C.L.) in the ΛCDM+r model. These new

Characterization of 16S rRNA Processing with Pre-30S Subunit Assembly Intermediates from E. coli.

PubMed

Smith, Brian A; Gupta, Neha; Denny, Kevin; Culver, Gloria M

2018-06-08

Ribosomal RNA (rRNA) is a major component of ribosomes and is fundamental to the process of translation. In bacteria, 16S rRNA is a component of the small ribosomal subunit and plays a critical role in mRNA decoding. rRNA maturation entails the removal of intervening spacer sequences contained within the pre-rRNA transcript by nucleolytic enzymes. Enzymatic activities involved in maturation of the 5'-end of 16S rRNA have been identified, but those involved in 3'-end maturation of 16S rRNA are more enigmatic. Here, we investigate molecular details of 16S rRNA maturation using purified in vivo-formed small subunit (SSU) assembly intermediates (pre-SSUs) from wild-type Escherichia coli that contain precursor 16S rRNA (17S rRNA). Upon incubation of pre-SSUs with E. coli S100 cell extracts or purified enzymes implicated in 16S rRNA processing, the 17S rRNA is processed into additional intermediates and mature 16S rRNA. These results illustrate that exonucleases RNase R, RNase II, PNPase, and RNase PH can process the 3'-end of pre-SSUs in vitro. However, the endonuclease YbeY did not exhibit nucleolytic activity with pre-SSUs under these conditions. Furthermore, these data demonstrate that multiple pathways facilitate 16S rRNA maturation with pre-SSUs in vitro, with the dominant pathways entailing complete processing of the 5'-end of 17S rRNA prior to 3'-end maturation or partial processing of the 5'-end with concomitant processing of the 3'-end. These results reveal the multifaceted nature of SSU biogenesis and suggest that E. coli may be able to escape inactivation of any one enzyme by using an existing complementary pathway. Copyright © 2018 Elsevier Ltd. All rights reserved.

Modular Extensions of Unitary Braided Fusion Categories and 2+1D Topological/SPT Orders with Symmetries

NASA Astrophysics Data System (ADS)

Lan, Tian; Kong, Liang; Wen, Xiao-Gang

2017-04-01

A finite bosonic or fermionic symmetry can be described uniquely by a symmetric fusion category E. In this work, we propose that 2+1D topological/SPT orders with a fixed finite symmetry E are classified, up to {E_8} quantum Hall states, by the unitary modular tensor categories C over E and the modular extensions of each C. In the case C=E, we prove that the set M_{ext}(E) of all modular extensions of E has a natural structure of a finite abelian group. We also prove that the set M_{ext}(C) of all modular extensions of E, if not empty, is equipped with a natural M_{ext}(C)-action that is free and transitive. Namely, the set M_{ext}(C) is an M_{ext}(E)-torsor. As special cases, we explain in detail how the group M_{ext}(E) recovers the well-known group-cohomology classification of the 2+1D bosonic SPT orders and Kitaev's 16 fold ways. We also discuss briefly the behavior of the group M_{ext}(E) under the symmetry-breaking processes and its relation to Witt groups.

Development of specification for the superpave simple performance tests (SPT).

DOT National Transportation Integrated Search

2009-05-16

This report describes the development and establishment of a proposed Simple Performance : Test (SPT) specification in order to contribute to the asphalt materials technology in the state of : Michigan. The properties and characteristic of materials,...

16 CFR 460.13 - Fact sheets.

Code of Federal Regulations, 2012 CFR

2012-01-01

... heat flow. The higher the R-value, the greater the insulating power. Compare insulation R-values before... INSULATION § 460.13 Fact sheets. If you are a manufacturer, you must give retailers and installers fact sheets for the insulation products you sell to them. Each sheet must contain what is listed here. You can...

16 CFR 460.13 - Fact sheets.

Code of Federal Regulations, 2010 CFR

2010-01-01

... heat flow. The higher the R-value, the greater the insulating power. Compare insulation R-values before... INSULATION § 460.13 Fact sheets. If you are a manufacturer, you must give retailers and installers fact sheets for the insulation products you sell to them. Each sheet must contain what is listed here. You can...

16 CFR 460.13 - Fact sheets.

Code of Federal Regulations, 2014 CFR

2014-01-01

... heat flow. The higher the R-value, the greater the insulating power. Compare insulation R-values before... INSULATION § 460.13 Fact sheets. If you are a manufacturer, you must give retailers and installers fact sheets for the insulation products you sell to them. Each sheet must contain what is listed here. You can...

16 CFR 460.13 - Fact sheets.

Code of Federal Regulations, 2013 CFR

2013-01-01

... heat flow. The higher the R-value, the greater the insulating power. Compare insulation R-values before... INSULATION § 460.13 Fact sheets. If you are a manufacturer, you must give retailers and installers fact sheets for the insulation products you sell to them. Each sheet must contain what is listed here. You can...

16 CFR 460.13 - Fact sheets.

Code of Federal Regulations, 2011 CFR

2011-01-01

... heat flow. The higher the R-value, the greater the insulating power. Compare insulation R-values before... INSULATION § 460.13 Fact sheets. If you are a manufacturer, you must give retailers and installers fact sheets for the insulation products you sell to them. Each sheet must contain what is listed here. You can...

Mediator Recruitment to Heat Shock Genes Requires Dual Hsf1 Activation Domains and Mediator Tail Subunits Med15 and Med16*

PubMed Central

Kim, Sunyoung; Gross, David S.

2013-01-01

The evolutionarily conserved Mediator complex is central to the regulation of gene transcription in eukaryotes because it serves as a physical and functional interface between upstream regulators and the Pol II transcriptional machinery. Nonetheless, its role appears to be context-dependent, and the detailed mechanism by which it governs the expression of most genes remains unknown. Here we investigate Mediator involvement in HSP (heat shock protein) gene regulation in the yeast Saccharomyces cerevisiae. We find that in response to thermal upshift, subunits representative of each of the four Mediator modules (Head, Middle, Tail, and Kinase) are rapidly, robustly, and selectively recruited to the promoter regions of HSP genes. Their residence is transient, returning to near-background levels within 90 min. Hsf1 (heat shock factor 1) plays a central role in recruiting Mediator, as indicated by the fact that truncation of either its N- or C-terminal activation domain significantly reduces Mediator occupancy, whereas removal of both activation domains abolishes it. Likewise, ablation of either of two Mediator Tail subunits, Med15 or Med16, reduces Mediator recruitment to HSP promoters, whereas deletion of both abolishes it. Accompanying the loss of Mediator, recruitment of RNA polymerase II is substantially diminished. Interestingly, Mediator antagonizes Hsf1 occupancy of non-induced promoters yet facilitates enhanced Hsf1 association with activated ones. Collectively, our observations indicate that Hsf1, via its dual activation domains, recruits holo-Mediator to HSP promoters in response to acute heat stress through cooperative physical and/or functional interactions with the Tail module. PMID:23447536

NASA's Hubble Spots Embryonic Galaxy SPT0615-JD

NASA Image and Video Library

2018-01-11

This Hubble Space Telescope image shows the farthest galaxy yet seen in an image that has been stretched and amplified by a phenomenon called gravitational lensing. The embryonic galaxy, named SPT0615-JD, existed when the universe was just 500 million years old. Though a few other primitive galaxies have been seen at this early epoch, they have essentially all looked like red dots, given their small size and tremendous distances. However, in this case, the gravitational field of a massive foreground galaxy cluster, called SPT-CL J0615-5746, not only amplified the light from the background galaxy but also smeared the image of it into an arc (about 2 arcseconds long). Image analysis shows that the galaxy weighs in at no more than 3 billion solar masses (roughly 1/100th the mass of our fully grown Milky Way galaxy). It is less than 2,500 light-years across, half the size of the Small Magellanic Cloud, a satellite galaxy of our Milky Way. The object is considered prototypical of young galaxies that emerged during the epoch shortly after the big bang. https://photojournal.jpl.nasa.gov/catalog/PIA22079

Analysis of Subcellular RNA Fractions Revealed a Transcription-Independent Effect of Tumor Necrosis Factor Alpha on Splicing, Mediated by Spt5.

PubMed

Diamant, Gil; Eisenbaum, Tal; Leshkowitz, Dena; Dikstein, Rivka

2016-05-01

The proinflammatory cytokine tumor necrosis factor alpha (TNF-α) modulates the expression of many genes, primarily through activation of NF-κB. Here, we examined the global effects of the elongation factor Spt5 on nascent and mature mRNAs of TNF-α-induced cells using chromatin and cytosolic subcellular fractions. We identified several classes of TNF-α-induced genes controlled at the level of transcription, splicing, and chromatin retention. Spt5 was found to facilitate splicing and chromatin release in genes displaying high induction rates. Further analysis revealed striking effects of TNF-α on the splicing of 25% of expressed genes; the vast majority were not transcriptionally induced. Splicing enhancement of noninduced genes by TNF-α was transient and independent of NF-κB. Investigating the underlying basis, we found that Spt5 is required for the splicing facilitation of the noninduced genes. In line with this, Spt5 interacts with Sm core protein splicing factors. Furthermore, following TNF-α treatment, levels of RNA polymerase II (Pol II) but not Spt5 are reduced from the splicing-induced genes, suggesting that these genes become enriched with a Pol II-Spt5 form. Our findings revealed the Pol II-Spt5 complex as a highly competent coordinator of cotranscriptional splicing. Copyright © 2016, American Society for Microbiology. All Rights Reserved.

TMEM16A is associated with voltage-gated calcium channels in mouse retina and its function is disrupted upon mutation of the auxiliary α2δ4 subunit

PubMed Central

Caputo, Antonella; Piano, Ilaria; Demontis, Gian Carlo; Bacchi, Niccolò; Casarosa, Simona; Santina, Luca Della; Gargini, Claudia

2015-01-01

Photoreceptors rely upon highly specialized synapses to efficiently transmit signals to multiple postsynaptic targets. Calcium influx in the presynaptic terminal is mediated by voltage-gated calcium channels (VGCC). This event triggers neurotransmitter release, but also gates calcium-activated chloride channels (TMEM), which in turn regulate VGCC activity. In order to investigate the relationship between VGCC and TMEM channels, we analyzed the retina of wild type (WT) and Cacna2d4 mutant mice, in which the VGCC auxiliary α2δ4 subunit carries a nonsense mutation, disrupting the normal channel function. Synaptic terminals of mutant photoreceptors are disarranged and synaptic proteins as well as TMEM16A channels lose their characteristic localization. In parallel, calcium-activated chloride currents are impaired in rods, despite unaltered TMEM16A protein levels. Co-immunoprecipitation revealed the interaction between VGCC and TMEM16A channels in the retina. Heterologous expression of these channels in tsA-201 cells showed that TMEM16A associates with the CaV1.4 subunit, and the association persists upon expression of the mutant α2δ4 subunit. Collectively, our experiments show association between TMEM16A and the α1 subunit of VGCC. Close proximity of these channels allows optimal function of the photoreceptor synaptic terminal under physiological conditions, but also makes TMEM16A channels susceptible to changes occurring to calcium channels. PMID:26557056

Highly conserved small subunit residues influence rubisco large subunit catalysis.

PubMed

Genkov, Todor; Spreitzer, Robert J

2009-10-30

The chloroplast enzyme ribulose 1,5-bisphosphate carboxylase/oxygenase (Rubisco) catalyzes the rate-limiting step of photosynthetic CO(2) fixation. With a deeper understanding of its structure-function relationships and competitive inhibition by O(2), it may be possible to engineer an increase in agricultural productivity and renewable energy. The chloroplast-encoded large subunits form the active site, but the nuclear-encoded small subunits can also influence catalytic efficiency and CO(2)/O(2) specificity. To further define the role of the small subunit in Rubisco function, the 10 most conserved residues in all small subunits were substituted with alanine by transformation of a Chlamydomonas reinhardtii mutant that lacks the small subunit gene family. All the mutant strains were able to grow photosynthetically, indicating that none of the residues is essential for function. Three of the substitutions have little or no effect (S16A, P19A, and E92A), one primarily affects holoenzyme stability (L18A), and the remainder affect catalysis with or without some level of associated structural instability (Y32A, E43A, W73A, L78A, P79A, and F81A). Y32A and E43A cause decreases in CO(2)/O(2) specificity. Based on the x-ray crystal structure of Chlamydomonas Rubisco, all but one (Glu-92) of the conserved residues are in contact with large subunits and cluster near the amino- or carboxyl-terminal ends of large subunit alpha-helix 8, which is a structural element of the alpha/beta-barrel active site. Small subunit residues Glu-43 and Trp-73 identify a possible structural connection between active site alpha-helix 8 and the highly variable small subunit loop between beta-strands A and B, which can also influence Rubisco CO(2)/O(2) specificity.

Maps of the Southern Millimeter-wave Sky from Combined 2500 deg$^2$ SPT-SZ and Planck Temperature Data

DOE Office of Scientific and Technical Information (OSTI.GOV)

Chown, R.; et al.

We present three maps of the millimeter-wave sky created by combining data from the South Pole Telescope (SPT) and the Planck satellite. We use data from the SPT-SZ survey, a survey of 2540 deg$^2$ of the the sky with arcminute resolution in three bands centered at 95, 150, and 220 GHz, and the full-mission Planck temperature data in the 100, 143, and 217 GHz bands. A linear combination of the SPT-SZ and Planck data is computed in spherical harmonic space, with weights derived from the noise of both instruments. This weighting scheme results in Planck data providing most of themore » large-angular-scale information in the combined maps, with the smaller-scale information coming from SPT-SZ data. A number of tests have been done on the maps. We find their angular power spectra to agree very well with theoretically predicted spectra and previously published results.« less

Cross-correlation of gravitational lensing from DES Science Verification data with SPT and Planck lensing

DOE Office of Scientific and Technical Information (OSTI.GOV)

Kirk, D.; Omori, Y.; Benoit-Lévy, A.

We measure the cross-correlation between weak lensing of galaxy images and of the cosmic microwave background (CMB). The effects of gravitational lensing on different sources will be correlated if the lensing is caused by the same mass fluctuations. We use galaxy shape measurements from 139 deg(2) of the Dark Energy Survey (DES) Science Verification data and overlapping CMB lensing from the South Pole Telescope (SPT) and Planck. The DES source galaxies have a median redshift of z(med) similar to 0.7, while the CMB lensing kernel is broad and peaks at z similar to 2. The resulting cross-correlation is maximally sensitivemore » to mass fluctuations at z similar to 0.44. Assuming the Planck 2015 best-fitting cosmology, the amplitude of the DESxSPT cross-power is found to be A(SPT) = 0.88 +/- 0.30 and that from DESxPlanck to be A(Planck) = 0.86 +/- 0.39, where A = 1 corresponds to the theoretical prediction. These are consistent with the expected signal and correspond to significances of 2.9 sigma and 2.2 sigma, respectively. We demonstrate that our results are robust to a number of important systematic effects including the shear measurement method, estimator choice, photo-z uncertainty and CMB lensing systematics. We calculate a value of A = 1.08 +/- 0.36 for DESxSPT when we correct the observations with a simple intrinsic alignment model. With three measurements of this cross-correlation now existing in the literature, there is not yet reliable evidence for any deviation from the expected LCDM level of cross-correlation. We provide forecasts for the expected signal-to-noise ratio of the combination of the five-year DES survey and SPT-3G.« less

Overexpression of stress-inducible OsBURP16, the β subunit of polygalacturonase 1, decreases pectin content and cell adhesion and increases abiotic stress sensitivity in rice

PubMed Central

Liu, Huanhuan; Ma, Yan; Chen, Na; Guo, Siyi; Liu, Huili; Guo, Xiaoyu; Chong, Kang; Xu, Yunyuan

2014-01-01

Polygalacturonase (PG), one of the hydrolases responsible for cell wall pectin degradation, is involved in organ consenescence and biotic stress in plants. PG1 is composed of a catalytic subunit, PG2, and a non-catalytic PG1β subunit. OsBURP16 belongs to the PG1β-like subfamily of BURP-family genes and encodes one putative PG1β subunit precursor in rice (Oryza sativa L.). Transcription of OsBURP16 is induced by cold, salinity and drought stresses, as well as by abscisic acid (ABA) treatment. Analysis of plant survival rates, relative ion leakage rates, accumulation levels of H2O2 and water loss rates of leaves showed that overexpression of OsBURP16 enhanced sensitivity to cold, salinity and drought stresses compared with controls. Young leaves of Ubi::OsBURP16 transgenic plants showed reduced cell adhesion and increased cuticular transpiration rate. Mechanical strength measurement of Ubi::OsBURP16 plants showed that reduced force was required to break leaves as compared with wild type. Transgenic rice showed enhanced PG activity and reduced pectin content. All these results suggested that overexpression of OsBURP16 caused pectin degradation and affected cell wall integrity as well as transpiration rate, which decreased tolerance to abiotic stresses. The cell wall is a barrier against biotic and abiotic stresses. Overexpression of stress-inducible OsBURP16, the beta-subunit of polygalacturonase 1, decreases pectin contents and cell adhesion in rice. Analyses of plant survival, ion leakage, H2O2 levels, and leaf water loss showed that these effects of overexpression were accompanied by enhanced sensitivity to cold, salinity and drought compared to the wild-type. Our data therefore provide new information on links between polygalacturonase activity and abiotic stress resistance in rice. PMID:24237159

Structure–function studies of histone H3/H4 tetramer maintenance during transcription by chaperone Spt2

DOE Office of Scientific and Technical Information (OSTI.GOV)

Chen, Shoudeng; Rufiange, Anne; Huang, Hongda

Cells use specific mechanisms such as histone chaperones to abrogate the inherent barrier that the nucleosome poses to transcribing polymerases. The current model postulates that nucleosomes can be transiently disrupted to accommodate passage of RNA polymerases and that histones H3 and H4 possess their own chaperones dedicated to the recovery of nucleosomes. Here, we determined the crystal structure of the conserved C terminus of human Suppressors of Ty insertions 2 (hSpt2C) chaperone bound to an H3/H4 tetramer. The structural studies demonstrate that hSpt2C is bound to the periphery of the H3/H4 tetramer, mimicking the trajectory of nucleosomal-bound DNA. These structuralmore » studies have been complemented with in vitro binding and in vivo functional studies on mutants that disrupt key intermolecular contacts involving two acidic patches and hydrophobic residues on Spt2C. We show that contacts between both human and yeast Spt2C with the H3/H4 tetramer are required for the suppression of H3/ H4 exchange as measured by H3K56ac and new H3 deposition. Furthermore, these interactions are also crucial for the inhibition of spurious transcription from within coding regions. In conclusion, together, our data indicate that Spt2 interacts with the periphery of the H3/H4 tetramer and promotes its recycling in the wake of RNA polymerase.« less

Structure–function studies of histone H3/H4 tetramer maintenance during transcription by chaperone Spt2

DOE PAGES

Chen, Shoudeng; Rufiange, Anne; Huang, Hongda; ...

2015-06-15

Cells use specific mechanisms such as histone chaperones to abrogate the inherent barrier that the nucleosome poses to transcribing polymerases. The current model postulates that nucleosomes can be transiently disrupted to accommodate passage of RNA polymerases and that histones H3 and H4 possess their own chaperones dedicated to the recovery of nucleosomes. Here, we determined the crystal structure of the conserved C terminus of human Suppressors of Ty insertions 2 (hSpt2C) chaperone bound to an H3/H4 tetramer. The structural studies demonstrate that hSpt2C is bound to the periphery of the H3/H4 tetramer, mimicking the trajectory of nucleosomal-bound DNA. These structuralmore » studies have been complemented with in vitro binding and in vivo functional studies on mutants that disrupt key intermolecular contacts involving two acidic patches and hydrophobic residues on Spt2C. We show that contacts between both human and yeast Spt2C with the H3/H4 tetramer are required for the suppression of H3/ H4 exchange as measured by H3K56ac and new H3 deposition. Furthermore, these interactions are also crucial for the inhibition of spurious transcription from within coding regions. In conclusion, together, our data indicate that Spt2 interacts with the periphery of the H3/H4 tetramer and promotes its recycling in the wake of RNA polymerase.« less

Improvement of Lead Tolerance of Saccharomyces cerevisiae by Random Mutagenesis of Transcription Regulator SPT3.

PubMed

Zhu, Liying; Gao, Shan; Zhang, Hongman; Huang, He; Jiang, Ling

2018-01-01

Bioremediation of heavy metal pollution with biomaterials such as bacteria and fungi usually suffer from limitations because of microbial sensitivity to high concentration of heavy metals. Herein, we adopted a novel random mutagenesis technique called RAISE to manipulate the transcription regulator SPT3 of Saccharomyces cerevisiae to improve cell lead tolerance. The best strain Mutant VI was selected from the random mutagenesis libraries on account of the growth performance, with higher specific growth rate than the control strain (0.068 vs. 0.040 h -1 ) at lead concentration as high as 1.8 g/L. Combined with the transcriptome analysis of S. cerevisiae, expressing the SPT3 protein was performed to make better sense of the global regulatory effects of SPT3. The data analysis revealed that 57 of S. cerevisiae genes were induced and 113 genes were suppressed, ranging from those for trehalose synthesis, carbon metabolism, and nucleotide synthesis to lead resistance. Especially, the accumulation of intracellular trehalose in S. cerevisiae under certain conditions of stress is considered important to lead resistance. The above results represented that SPT3 was acted as global transcription regulator in the exponential phase of strain and accordingly improved heavy metal tolerance in the heterologous host S. cerevisiae. The present study provides a route to complex phenotypes that are not readily accessible by traditional methods.

[Protective activity of S-PT84, a heat-killed preparation of Lactobacillus pentosus, against oral and gastric candidiasis in an experimental murine model].

PubMed

Hayama, Kazumi; Ishijima, Sanae; Ono, Yoshiko; Izumo, Takayuki; Ida, Masayuki; Shibata, Hiroshi; Abe, Shigeru

2014-01-01

The effect of S-PT84, a heat-killed preparation of Lactobacillus pentosus on growth of Candida albicans was examined in vitro and in vivo. The mycelial growth was effectively inhibited by S-PT84 and seemed to bind to the hyphae. We assessed the potential of S-PT84 for treatment of oral and gastric candidiasis using a murine model. When 2 mg of S-PT84 was administered three times into the oral cavity of orally Candida infected mice, the score of lesions on the tongue was improved on day 2. When 50 μl and 200 μl of S-PT84 (10 mg/ml) were administered three times into the oral cavity (0.5 mg × 3) and the stomach (2 mg × 3) of the same mouse model, the number of viable Candida cells in the stomach was reduced significantly on day 2. These findings suggest the possibility that S-PT84 has potential as a food ingredient supporting anti-Candida treatment, especially for Candida infection in the gastrointestinal tract.

Overexpression of stress-inducible OsBURP16, the β subunit of polygalacturonase 1, decreases pectin content and cell adhesion and increases abiotic stress sensitivity in rice.

PubMed

Liu, Huanhuan; Ma, Yan; Chen, Na; Guo, Siyi; Liu, Huili; Guo, Xiaoyu; Chong, Kang; Xu, Yunyuan

2014-05-01

Polygalacturonase (PG), one of the hydrolases responsible for cell wall pectin degradation, is involved in organ consenescence and biotic stress in plants. PG1 is composed of a catalytic subunit, PG2, and a non-catalytic PG1β subunit. OsBURP16 belongs to the PG1β-like subfamily of BURP-family genes and encodes one putative PG1β subunit precursor in rice (Oryza sativa L.). Transcription of OsBURP16 is induced by cold, salinity and drought stresses, as well as by abscisic acid (ABA) treatment. Analysis of plant survival rates, relative ion leakage rates, accumulation levels of H2 O2 and water loss rates of leaves showed that overexpression of OsBURP16 enhanced sensitivity to cold, salinity and drought stresses compared with controls. Young leaves of Ubi::OsBURP16 transgenic plants showed reduced cell adhesion and increased cuticular transpiration rate. Mechanical strength measurement of Ubi::OsBURP16 plants showed that reduced force was required to break leaves as compared with wild type. Transgenic rice showed enhanced PG activity and reduced pectin content. All these results suggested that overexpression of OsBURP16 caused pectin degradation and affected cell wall integrity as well as transpiration rate, which decreased tolerance to abiotic stresses. © 2013 The Authors. Plant, Cell & Environment published by John Wiley & Sons Ltd.

Architecture of the Saccharomyces cerevisiae SAGA transcription coactivator complex

PubMed Central

Han, Yan; Luo, Jie; Ranish, Jeffrey; Hahn, Steven

2014-01-01

The conserved transcription coactivator SAGA is comprised of several modules that are involved in activator binding, TBP binding, histone acetylation (HAT) and deubiquitination (DUB). Crosslinking and mass spectrometry, together with genetic and biochemical analyses, were used to determine the molecular architecture of the SAGA-TBP complex. We find that the SAGA Taf and Taf-like subunits form a TFIID-like core complex at the center of SAGA that makes extensive interactions with all other SAGA modules. SAGA-TBP binding involves a network of interactions between subunits Spt3, Spt8, Spt20, and Spt7. The HAT and DUB modules are in close proximity, and the DUB module modestly stimulates HAT function. The large activator-binding subunit Tra1 primarily connects to the TFIID-like core via its FAT domain. These combined results were used to derive a model for the arrangement of the SAGA subunits and its interactions with TBP. Our results provide new insight into SAGA function in gene regulation, its structural similarity with TFIID, and functional interactions between the SAGA modules. PMID:25216679

Neighborhood of 16S rRNA nucleotides U788/U789 in the 30S ribosomal subunit determined by site-directed crosslinking.

PubMed

Mundus, D; Wollenzien, P

1998-11-01

Site-specific photo crosslinking has been used to investigate the RNA neighborhood of 16S rRNA positions U788/ U789 in Escherichia coli 30S subunits. For these studies, site-specific psoralen (SSP) which contains a sulfhydryl group on a 17 A side chain was first added to nucleotides U788/U789 using a complementary guide DNA by annealing and phototransfer. Modified RNA was purified from the DNA and unmodified RNA. For some experiments, the SSP, which normally crosslinks at an 8 A distance, was derivitized with azidophenacylbromide (APAB) resulting in the photoreactive azido moiety at a maximum of 25 A from the 4' position on psoralen (SSP25APA). 16S rRNA containing SSP, SSP25APA or control 16S rRNA were reconstituted and 30S particles were isolated. The reconstituted subunits containing SSP or SSP25APA had normal protein composition, were active in tRNA binding and had the usual pattern of chemical reactivity except for increased kethoxal reactivity at G791 and modest changes in four other regions. Irradiation of the derivatized 30S subunits in activation buffer produced several intramolecular RNA crosslinks that were visualized and separated by gel electrophoresis and characterized by primer extension. Four major crosslink sites made by the SSP reagent were identified at positions U561/U562, U920/U921, C866 and U723; a fifth major crosslink at G693 was identified when the SSP25APA reagent was used. A number of additional crosslinks of lower frequency were seen, particularly with the APA reagent. These data indicate a central location close to the decoding region and central pseudoknot for nucleotides U788/U789 in the activated 30S subunit.

16 CFR 5.63 - Evidence; transcript; in camera orders; proposed findings of fact and conclusions of law.

Code of Federal Regulations, 2010 CFR

2010-01-01

... 16 Commercial Practices 1 2010-01-01 2010-01-01 false Evidence; transcript; in camera orders; proposed findings of fact and conclusions of law. 5.63 Section 5.63 Commercial Practices FEDERAL TRADE... findings of fact and conclusions of law. Sections 3.43, 3.44, 3.45, and 3.46 of the Commission's Rules of...

MASS CALIBRATION AND COSMOLOGICAL ANALYSIS OF THE SPT-SZ GALAXY CLUSTER SAMPLE USING VELOCITY DISPERSION σ {sub v} AND X-RAY Y {sub X} MEASUREMENTS

DOE Office of Scientific and Technical Information (OSTI.GOV)

Bocquet, S.; Saro, A.; Mohr, J. J.

2015-02-01

We present a velocity-dispersion-based mass calibration of the South Pole Telescope Sunyaev-Zel'dovich effect survey (SPT-SZ) galaxy cluster sample. Using a homogeneously selected sample of 100 cluster candidates from 720 deg{sup 2} of the survey along with 63 velocity dispersion (σ {sub v}) and 16 X-ray Y {sub X} measurements of sample clusters, we simultaneously calibrate the mass-observable relation and constrain cosmological parameters. Our method accounts for cluster selection, cosmological sensitivity, and uncertainties in the mass calibrators. The calibrations using σ {sub v} and Y {sub X} are consistent at the 0.6σ level, with the σ {sub v} calibration preferring ∼16% highermore » masses. We use the full SPT{sub CL} data set (SZ clusters+σ {sub v}+Y {sub X}) to measure σ{sub 8}(Ω{sub m}/0.27){sup 0.3} = 0.809 ± 0.036 within a flat ΛCDM model. The SPT cluster abundance is lower than preferred by either the WMAP9 or Planck+WMAP9 polarization (WP) data, but assuming that the sum of the neutrino masses is ∑m {sub ν} = 0.06 eV, we find the data sets to be consistent at the 1.0σ level for WMAP9 and 1.5σ for Planck+WP. Allowing for larger ∑m {sub ν} further reconciles the results. When we combine the SPT{sub CL} and Planck+WP data sets with information from baryon acoustic oscillations and Type Ia supernovae, the preferred cluster masses are 1.9σ higher than the Y {sub X} calibration and 0.8σ higher than the σ {sub v} calibration. Given the scale of these shifts (∼44% and ∼23% in mass, respectively), we execute a goodness-of-fit test; it reveals no tension, indicating that the best-fit model provides an adequate description of the data. Using the multi-probe data set, we measure Ω{sub m} = 0.299 ± 0.009 and σ{sub 8} = 0.829 ± 0.011. Within a νCDM model we find ∑m {sub ν} = 0.148 ± 0.081 eV. We present a consistency test of the cosmic growth rate using SPT clusters. Allowing both the growth index γ and the dark energy

Identification of nucleotides in E. coli 16S rRNA essential for ribosome subunit association.

PubMed

Pulk, Arto; Maiväli, Ulo; Remme, Jaanus

2006-05-01

The ribosome consists of two unequal subunits, which associate via numerous intersubunit contacts. Medium-resolution structural studies have led to grouping of the intersubunit contacts into 12 directly visualizable intersubunit bridges. Most of the intersubunit interactions involve RNA. We have used an RNA modification interference approach to determine Escherichia coli 16S rRNA positions that are essential for the association of functionally active 70S ribosomes. Modification of the N1 position of A702, A1418, and A1483 with DMS, and of the N3 position of U793, U1414, and U1495 with CMCT in 30S subunits strongly interferes with 70S ribosome formation. Five of these positions localize into previously recognized intersubunit bridges, namely, B2a (U1495), B2b (U793), B3 (A1483), B5 (A1418), and B7a (A702). The remaining position displaying interference, U1414, forms a base pair with G1486, which is a part of bridge B3. We contend that these five intersubunit bridges are essential for reassociation of the 70S ribosome, thus forming the functional core of the intersubunit contacts.

Identification of nucleotides in E. coli 16S rRNA essential for ribosome subunit association

PubMed Central

Pulk, Arto; Maiväli, Ülo; Remme, Jaanus

2006-01-01

The ribosome consists of two unequal subunits, which associate via numerous intersubunit contacts. Medium-resolution structural studies have led to grouping of the intersubunit contacts into 12 directly visualizable intersubunit bridges. Most of the intersubunit interactions involve RNA. We have used an RNA modification interference approach to determine Escherichia coli 16S rRNA positions that are essential for the association of functionally active 70S ribosomes. Modification of the N1 position of A702, A1418, and A1483 with DMS, and of the N3 position of U793, U1414, and U1495 with CMCT in 30S subunits strongly interferes with 70S ribosome formation. Five of these positions localize into previously recognized intersubunit bridges, namely, B2a (U1495), B2b (U793), B3 (A1483), B5 (A1418), and B7a (A702). The remaining position displaying interference, U1414, forms a base pair with G1486, which is a part of bridge B3. We contend that these five intersubunit bridges are essential for reassociation of the 70S ribosome, thus forming the functional core of the intersubunit contacts. PMID:16556933

Large arrays of dual-polarized multichroic TES detectors for CMB measurements with the SPT-3G receiver

DOE PAGES

Holland, Wayne S.; Zmuidzinas, Jonas; Posada, Chrystian M.; ...

2016-07-19

Now, detectors for cosmic microwave background (CMB) experiments are background limited, so a straightforward alternative to improve sensitivity is to increase the number of detectors. Large arrays of multichroic pixels constitute an economical approach to increasing the number of detectors within a given focal plane area. We present the fabrication of large arrays of dual-polarized multichroic transition-edge-sensor (TES) bolometers for the South Pole Telescope third-generation CMB receiver (SPT-3G). The complete SPT-3G receiver will have 2690 pixels, each with six detectors, allowing for individual measurement of three spectral bands (centered at 95 GHz, 150 GHz and 220 GHz) in two orthogonalmore » polarizations. In total, the SPT-3G focal plane will have 16140 detectors. Each pixel is comprised of a broad-band sinuous antenna coupled to a niobium microstrip transmission line. In-line filters are used to define the different band-passes before the millimeter-wavelength signal is fed to the respective Ti/Au TES sensors. Detectors are read out using a 64x frequency domain multiplexing (fMux) scheme. The microfabrication of the SPT-3G detector arrays involves a total of 18 processes, including 13 lithography steps. Together with the fabrication process, the effect of processing on the Ti/Au TES's T-c is discussed. In addition, detectors fabricated with Ti/Au TES films with Tc between 400 mK 560 mK are presented and their thermal characteristics are evaluated. Optical characterization of the arrays is presented as well, indicating that the response of the detectors is in good agreement with the design values for all three spectral bands (95 GHz, 150 GHz, and 220 GHz). The measured optical efficiency of the detectors is between 0.3 and 0.8. Our results discussed here are extracted from a batch of research of development wafers used to develop the baseline process for the fabrication of the arrays of detectors to be deployed with the SPT-3G receiver. Results from

Large arrays of dual-polarized multichroic TES detectors for CMB measurements with the SPT-3G receiver

DOE Office of Scientific and Technical Information (OSTI.GOV)

Holland, Wayne S.; Zmuidzinas, Jonas; Posada, Chrystian M.

Now, detectors for cosmic microwave background (CMB) experiments are background limited, so a straightforward alternative to improve sensitivity is to increase the number of detectors. Large arrays of multichroic pixels constitute an economical approach to increasing the number of detectors within a given focal plane area. We present the fabrication of large arrays of dual-polarized multichroic transition-edge-sensor (TES) bolometers for the South Pole Telescope third-generation CMB receiver (SPT-3G). The complete SPT-3G receiver will have 2690 pixels, each with six detectors, allowing for individual measurement of three spectral bands (centered at 95 GHz, 150 GHz and 220 GHz) in two orthogonalmore » polarizations. In total, the SPT-3G focal plane will have 16140 detectors. Each pixel is comprised of a broad-band sinuous antenna coupled to a niobium microstrip transmission line. In-line filters are used to define the different band-passes before the millimeter-wavelength signal is fed to the respective Ti/Au TES sensors. Detectors are read out using a 64x frequency domain multiplexing (fMux) scheme. The microfabrication of the SPT-3G detector arrays involves a total of 18 processes, including 13 lithography steps. Together with the fabrication process, the effect of processing on the Ti/Au TES's T-c is discussed. In addition, detectors fabricated with Ti/Au TES films with Tc between 400 mK 560 mK are presented and their thermal characteristics are evaluated. Optical characterization of the arrays is presented as well, indicating that the response of the detectors is in good agreement with the design values for all three spectral bands (95 GHz, 150 GHz, and 220 GHz). The measured optical efficiency of the detectors is between 0.3 and 0.8. Our results discussed here are extracted from a batch of research of development wafers used to develop the baseline process for the fabrication of the arrays of detectors to be deployed with the SPT-3G receiver. Results from

Large arrays of dual-polarized multichroic TES detectors for CMB measurements with the SPT-3G receiver

NASA Astrophysics Data System (ADS)

Posada, Chrystian M.; Ade, Peter A. R.; Anderson, Adam J.; Avva, Jessica; Ahmed, Zeeshan; Arnold, Kam S.; Austermann, Jason; Bender, Amy N.; Benson, Bradford A.; Bleem, Lindsey; Byrum, Karen; Carlstrom, John E.; Carter, Faustin W.; Chang, Clarence; Cho, Hsiao-Mei; Cukierman, Ari; Czaplewski, David A.; Ding, Junjia; Divan, Ralu N. S.; de Haan, Tijmen; Dobbs, Matt; Dutcher, Daniel; Everett, Wenderline; Gannon, Renae N.; Guyser, Robert J.; Halverson, Nils W.; Harrington, Nicholas L.; Hattori, Kaori; Henning, Jason W.; Hilton, Gene C.; Holzapfel, William L.; Huang, Nicholas; Irwin, Kent D.; Jeong, Oliver; Khaire, Trupti; Korman, Milo; Kubik, Donna L.; Kuo, Chao-Lin; Lee, Adrian T.; Leitch, Erik M.; Lendinez Escudero, Sergi; Meyer, Stephan S.; Miller, Christina S.; Montgomery, Joshua; Nadolski, Andrew; Natoli, Tyler J.; Nguyen, Hogan; Novosad, Valentyn; Padin, Stephen; Pan, Zhaodi; Pearson, John E.; Rahlin, Alexandra; Reichardt, Christian L.; Ruhl, John E.; Saliwanchik, Benjamin; Shirley, Ian; Sayre, James T.; Shariff, Jamil A.; Shirokoff, Erik D.; Stan, Liliana; Stark, Antony A.; Sobrin, Joshua; Story, Kyle; Suzuki, Aritoki; Tang, Qing Yang; Thakur, Ritoban B.; Thompson, Keith L.; Tucker, Carole E.; Vanderlinde, Keith; Vieira, Joaquin D.; Wang, Gensheng; Whitehorn, Nathan; Yefremenko, Volodymyr; Yoon, Ki Won

2016-07-01

Detectors for cosmic microwave background (CMB) experiments are now essentially background limited, so a straightforward alternative to improve sensitivity is to increase the number of detectors. Large arrays of multichroic pixels constitute an economical approach to increasing the number of detectors within a given focal plane area. Here, we present the fabrication of large arrays of dual-polarized multichroic transition-edge-sensor (TES) bolometers for the South Pole Telescope third-generation CMB receiver (SPT-3G). The complete SPT-3G receiver will have 2690 pixels, each with six detectors, allowing for individual measurement of three spectral bands (centered at 95 GHz, 150 GHz and 220 GHz) in two orthogonal polarizations. In total, the SPT-3G focal plane will have 16140 detectors. Each pixel is comprised of a broad-band sinuous antenna coupled to a niobium microstrip transmission line. In-line filters are used to define the different band-passes before the millimeter-wavelength signal is fed to the respective Ti/Au TES sensors. Detectors are read out using a 64x frequency domain multiplexing (fMux) scheme. The microfabrication of the SPT-3G detector arrays involves a total of 18 processes, including 13 lithography steps. Together with the fabrication process, the effect of processing on the Ti/Au TES's Tc is discussed. In addition, detectors fabricated with Ti/Au TES films with Tc between 400 mK 560 mK are presented and their thermal characteristics are evaluated. Optical characterization of the arrays is presented as well, indicating that the response of the detectors is in good agreement with the design values for all three spectral bands (95 GHz, 150 GHz, and 220 GHz). The measured optical efficiency of the detectors is between 0.3 and 0.8. Results discussed here are extracted from a batch of research of development wafers used to develop the baseline process for the fabrication of the arrays of detectors to be deployed with the SPT-3G receiver. Results from

Effects of the β1 auxiliary subunit on modification of Rat Na{sub v}1.6 sodium channels expressed in HEK293 cells by the pyrethroid insecticides tefluthrin and deltamethrin

DOE Office of Scientific and Technical Information (OSTI.GOV)

He, Bingjun; Soderlund, David M., E-mail: dms6@cornell.edu

We expressed rat Na{sub v}1.6 sodium channels with or without the rat β1 subunit in human embryonic kidney (HEK293) cells and evaluated the effects of the pyrethroid insecticides tefluthrin and deltamethrin on whole-cell sodium currents. In assays with the Na{sub v}1.6 α subunit alone, both pyrethroids prolonged channel inactivation and deactivation and shifted the voltage dependence of channel activation and steady-state inactivation toward hyperpolarization. Maximal shifts in activation were ~ 18 mV for tefluthrin and ~ 24 mV for deltamethrin. These compounds also caused hyperpolarizing shifts of ~ 10–14 mV in the voltage dependence of steady-state inactivation and increased inmore » the fraction of sodium current that was resistant to inactivation. The effects of pyrethroids on the voltage-dependent gating greatly increased the size of sodium window currents compared to unmodified channels; modified channels exhibited increased probability of spontaneous opening at membrane potentials more negative than the normal threshold for channel activation and incomplete channel inactivation. Coexpression of Na{sub v}1.6 with the β1 subunit had no effect on the kinetic behavior of pyrethroid-modified channels but had divergent effects on the voltage-dependent gating of tefluthrin- or deltamethrin-modified channels, increasing the size of tefluthrin-induced window currents but decreasing the size of corresponding deltamethrin-induced currents. Unexpectedly, the β1 subunit did not confer sensitivity to use-dependent channel modification by either tefluthrin or deltamethrin. We conclude from these results that functional reconstitution of channels in vitro requires careful attention to the subunit composition of channel complexes to ensure that channels in vitro are faithful functional and pharmacological models of channels in neurons. - Highlights: • We expressed Na{sub v}1.6 sodium channels with or without β1 subunits in HEK293 cells. • Tefluthrin and

Cosmological constraints from Galaxy Clusters in 2500 square-degree SPT-SZ survey

DOE Office of Scientific and Technical Information (OSTI.GOV)

Haan, T. de; Benson, B. A.; Bleem, L. E.

We present cosmological parameter constraints obtained from galaxy clusters identified by their SunyaevZel'dovich effect signature in the 2500 square-degree South Pole Telescope Sunyaev Zel'dovich (SPT-SZ) survey. We consider the 377 cluster candidates identified at z > 0.25 with a detection significance greater than five, corresponding to the 95% purity threshold for the survey. We compute constraints on cosmological models using the measured cluster abundance as a function of mass and redshift. We include additional constraints from multi-wavelength observations, including Chandra X-ray data for 82 clusters and a weak lensing-based prior on the normalization of the mass-observable scaling relations. Assuming amore » spatially flat Lambda CDM cosmology, we combine the cluster data with a prior on H-0 and find sigma(8)= 0.784. +/- 0.039 and Omega(m) = 0.289. +/- 0.042, with the parameter combination sigma(8) (Omega(m)/0.27)(0.3) = 0.797 +/- 0.031. These results are in good agreement with constraints from the cosmic microwave background (CMB) from SPT, WMAP, and Planck, as well as with constraints from other cluster data sets. We also consider several extensions to Lambda CDM, including models in which the equation of state of dark energy w, the species-summed neutrino mass, and/or the effective number of relativistic species (N-eff) are free parameters. When combined with constraints from the Planck CMB, H-0, baryon acoustic oscillation, and SNe, adding the SPT cluster data improves the w constraint by 14%, to w = -1.023 +/- 0.042.« less

Tracking protons from respiratory chain complexes to ATP synthase c-subunit: The critical role of serine and threonine residues.

PubMed

Panfoli, Isabella; Ponassi, Marco; Ravera, Silvia; Calzia, Daniela; Beitia, Maider; Morelli, Alessandro; Rosano, Camillo

2017-01-22

F 1 F o -ATP synthase is a multisubunit enzyme responsible for the synthesis of ATP. Among its multiple subunits (8 in E. coli, 17 in yeast S. cerevisiae, 16 in vertebrates), two subunits a and c are known to play a central role controlling the H + flow through the inner mitochondrial membrane which allows the subsequent synthesis of ATP, but the pathway followed by H + within the two proteins is still a matter of debate. In fact, even though the structure of ATP synthase is now well defined, the molecular mechanisms determining the function of both F 1 and F O domains are still largely unknown. In this study, we propose a pathway for proton migration along the ATP synthase by hydrogen-bonded chain mechanism, with a key role of serine and threonine residues, by X-ray diffraction data on the subunit a of E. coli Fo. Copyright © 2016 Elsevier Inc. All rights reserved.

Tracking protons from respiratory chain complexes to ATP synthase c-subunit: The critical role of serine and threonine residues

DOE Office of Scientific and Technical Information (OSTI.GOV)

Panfoli, Isabella; Ponassi, Marco; Ravera, Silvia

F{sub 1}F{sub o}-ATP synthase is a multisubunit enzyme responsible for the synthesis of ATP. Among its multiple subunits (8 in E. coli, 17 in yeast S. cerevisiae, 16 in vertebrates), two subunits a and c are known to play a central role controlling the H{sup +} flow through the inner mitochondrial membrane which allows the subsequent synthesis of ATP, but the pathway followed by H{sup +} within the two proteins is still a matter of debate. In fact, even though the structure of ATP synthase is now well defined, the molecular mechanisms determining the function of both F{sub 1} andmore » F{sub O} domains are still largely unknown. In this study, we propose a pathway for proton migration along the ATP synthase by hydrogen-bonded chain mechanism, with a key role of serine and threonine residues, by X-ray diffraction data on the subunit a of E. coli Fo.« less

Pho dynamically interacts with Spt5 to facilitate transcriptional switches at the hsp70 locus.

PubMed

Pereira, Allwyn; Paro, Renato

2017-12-06

Numerous target genes of the Polycomb group (PcG) are transiently activated by a stimulus and subsequently repressed. However, mechanisms by which PcG proteins regulate such target genes remain elusive. We employed the heat shock-responsive hsp70 locus in Drosophila to study the chromatin dynamics of PRC1 and its interplay with known regulators of the locus before, during and after heat shock. We detected mutually exclusive binding patterns for HSF and PRC1 at the hsp70 locus. We found that Pleiohomeotic (Pho), a DNA-binding PcG member, dynamically interacts with Spt5, an elongation factor. The dynamic interaction switch between Pho and Spt5 is triggered by the recruitment of HSF to chromatin. Mutation in the protein-protein interaction domain (REPO domain) of Pho interferes with the dynamics of its interaction with Spt5. The transcriptional kinetics of the heat shock response is negatively affected by a mutation in the REPO domain of Pho. We propose that a dynamic interaction switch between PcG proteins and an elongation factor enables stress-inducible genes to efficiently switch between ON/OFF states in the presence/absence of the activating stimulus.

The Initiation Factor TFE and the Elongation Factor Spt4/5 Compete for the RNAP Clamp during Transcription Initiation and Elongation

PubMed Central

Grohmann, Dina; Nagy, Julia; Chakraborty, Anirban; Klose, Daniel; Fielden, Daniel; Ebright, Richard H.; Michaelis, Jens; Werner, Finn

2011-01-01

Summary TFIIE and the archaeal homolog TFE enhance DNA strand separation of eukaryotic RNAPII and the archaeal RNAP during transcription initiation by an unknown mechanism. We have developed a fluorescently labeled recombinant M. jannaschii RNAP system to probe the archaeal transcription initiation complex, consisting of promoter DNA, TBP, TFB, TFE, and RNAP. We have localized the position of the TFE winged helix (WH) and Zinc ribbon (ZR) domains on the RNAP using single-molecule FRET. The interaction sites of the TFE WH domain and the transcription elongation factor Spt4/5 overlap, and both factors compete for RNAP binding. Binding of Spt4/5 to RNAP represses promoter-directed transcription in the absence of TFE, which alleviates this effect by displacing Spt4/5 from RNAP. During elongation, Spt4/5 can displace TFE from the RNAP elongation complex and stimulate processivity. Our results identify the RNAP “clamp” region as a regulatory hot spot for both transcription initiation and transcription elongation. PMID:21777815

The equine LH/CGβ subunit combines divergent intracellular traits of the human LHβ and CGβ subunits

PubMed Central

Cohen, Limor; Bousfield, George R; Ben-Menahem, David

2017-01-01

The pituitary LHβ and placental CGβ subunits are products of different genes in primates. The major structural difference between the two subunits is in the carboxy-terminal region, where the short carboxyl sequence of hLHβ is replaced by a longer O-glycosylated carboxy-terminal peptide (CTP) in hCGβ. In association with this structural deviation, there are marked differences in the secretion kinetics and polarized routing of the two subunits. In equids, however, the CGβ and LHβ subunits are products of the same gene expressed in the placenta and pituitary (eLH/CGβ), and both contain a CTP. This unusual expression pattern intrigued us and led to our study of eLH/CGβ subunit secretion by transfected CHO and MDCK cells. In continuous labeling and pulse chase experiments, the secretion of the eLH/CGβ subunit from the transfected CHO cells was inefficient (medium recovery of 16–25%) and slow (t1/2 >6.5 hrs). This indicated that, the secretion of the eLH/CGβ subunit resembles that of hLHβ rather than hCGβ. In MDCK cells grown on Transwell filters, the eLH/CGβ subunit was preferentially secreted from the apical side, similar to the hCGβ subunit secretory route (~65% of the total protein secreted). Taken together, these data suggested that secretion of the eLH/CGβ subunit integrates features of both hLHβ and hCGβ subunits. We propose that the evolution of this intracellular behavior may fulfill the physiological demands for biosynthesis of the eLH/CGβ subunit in the pituitary as well as in the placenta. PMID:25796287

SPT0346-52: Negligible AGN Activity in a Compact, Hyper-starburst Galaxy at z = 5.7

NASA Astrophysics Data System (ADS)

Ma, Jingzhe; Gonzalez, Anthony. H.; Vieira, J. D.; Aravena, M.; Ashby, M. L. N.; Béthermin, M.; Bothwell, M. S.; Brandt, W. N.; de Breuck, C.; Carlstrom, J. E.; Chapman, S. C.; Gullberg, B.; Hezaveh, Y.; Litke, K.; Malkan, M.; Marrone, D. P.; McDonald, M.; Murphy, E. J.; Spilker, J. S.; Sreevani, J.; Stark, A. A.; Strandet, M.; Wang, S. X.

2016-12-01

We present Chandra ACIS-S and Australia Telescope Compact Array (ATCA) radio continuum observations of the strongly lensed dusty, star-forming galaxy SPT-S J034640-5204.9 (hereafter SPT0346-52) at z = 5.656. This galaxy has also been observed with ALMA, HST, Spitzer, Herschel, Atacama Pathfinder EXperiment, and the Very Large Telescope. Previous observations indicate that if the infrared (IR) emission is driven by star formation, then the inferred lensing-corrected star formation rate (SFR) (˜4500 M ⊙ yr-1) and SFR surface density ΣSFR (˜2000 M ⊙ yr-1 kpc-2) are both exceptionally high. It remained unclear from the previous data, however, whether a central active galactic nucleus (AGN) contributes appreciably to the IR luminosity. The Chandra upper limit shows that SPT0346-52 is consistent with being star formation dominated in the X-ray, and any AGN contribution to the IR emission is negligible. The ATCA radio continuum upper limits are also consistent with the FIR-to-radio correlation for star-forming galaxies with no indication of an additional AGN contribution. The observed prodigious intrinsic IR luminosity of (3.6 ± 0.3) × 1013 L ⊙ originates almost solely from vigorous star formation activity. With an intrinsic source size of 0.61 ± 0.03 kpc, SPT0346-52 is confirmed to have one of the highest ΣSFR of any known galaxy. This high ΣSFR, which approaches the Eddington limit for a radiation pressure supported starburst, may be explained by a combination of very high star formation efficiency and gas fraction.

Cross-correlation of gravitational lensing from DES Science Verification data with SPT and Planck lensing

NASA Astrophysics Data System (ADS)

Kirk, D.; Omori, Y.; Benoit-Lévy, A.; Cawthon, R.; Chang, C.; Larsen, P.; Amara, A.; Bacon, D.; Crawford, T. M.; Dodelson, S.; Fosalba, P.; Giannantonio, T.; Holder, G.; Jain, B.; Kacprzak, T.; Lahav, O.; MacCrann, N.; Nicola, A.; Refregier, A.; Sheldon, E.; Story, K. T.; Troxel, M. A.; Vieira, J. D.; Vikram, V.; Zuntz, J.; Abbott, T. M. C.; Abdalla, F. B.; Becker, M. R.; Benson, B. A.; Bernstein, G. M.; Bernstein, R. A.; Bleem, L. E.; Bonnett, C.; Bridle, S. L.; Brooks, D.; Buckley-Geer, E.; Burke, D. L.; Capozzi, D.; Carlstrom, J. E.; Rosell, A. Carnero; Kind, M. Carrasco; Carretero, J.; Crocce, M.; Cunha, C. E.; D'Andrea, C. B.; da Costa, L. N.; Desai, S.; Diehl, H. T.; Dietrich, J. P.; Doel, P.; Eifler, T. F.; Evrard, A. E.; Flaugher, B.; Frieman, J.; Gerdes, D. W.; Goldstein, D. A.; Gruen, D.; Gruendl, R. A.; Honscheid, K.; James, D. J.; Jarvis, M.; Kent, S.; Kuehn, K.; Kuropatkin, N.; Lima, M.; March, M.; Martini, P.; Melchior, P.; Miller, C. J.; Miquel, R.; Nichol, R. C.; Ogando, R.; Plazas, A. A.; Reichardt, C. L.; Roodman, A.; Rozo, E.; Rykoff, E. S.; Sako, M.; Sanchez, E.; Scarpine, V.; Schubnell, M.; Sevilla-Noarbe, I.; Simard, G.; Smith, R. C.; Soares-Santos, M.; Sobreira, F.; Suchyta, E.; Swanson, M. E. C.; Tarle, G.; Thomas, D.; Wechsler, R. H.; Weller, J.

2016-06-01

We measure the cross-correlation between weak lensing of galaxy images and of the cosmic microwave background (CMB). The effects of gravitational lensing on different sources will be correlated if the lensing is caused by the same mass fluctuations. We use galaxy shape measurements from 139 deg2 of the Dark Energy Survey (DES) Science Verification data and overlapping CMB lensing from the South Pole Telescope (SPT) and Planck. The DES source galaxies have a median redshift of zmed ˜ 0.7, while the CMB lensing kernel is broad and peaks at z ˜ 2. The resulting cross-correlation is maximally sensitive to mass fluctuations at z ˜ 0.44. Assuming the Planck 2015 best-fitting cosmology, the amplitude of the DES×SPT cross-power is found to be ASPT = 0.88 ± 0.30 and that from DES×Planck to be APlanck = 0.86 ± 0.39, where A = 1 corresponds to the theoretical prediction. These are consistent with the expected signal and correspond to significances of 2.9σ and 2.2σ, respectively. We demonstrate that our results are robust to a number of important systematic effects including the shear measurement method, estimator choice, photo-z uncertainty and CMB lensing systematics. We calculate a value of A = 1.08 ± 0.36 for DES×SPT when we correct the observations with a simple intrinsic alignment model. With three measurements of this cross-correlation now existing in the literature, there is not yet reliable evidence for any deviation from the expected LCDM level of cross-correlation. We provide forecasts for the expected signal-to-noise ratio of the combination of the five-year DES survey and SPT-3G.

The Subunit Principle in Scar Face Revision.

PubMed

Elshahat, Ahmed; Lashin, Riham

2017-06-01

Facial scaring is considered one of the most difficult cosmetic problems for any plastic surgeon to solve. The condition is more difficult if the direction of the scar is not parallel to relaxed skin tension lines. Attempts to manage this difficult situation included revisions using geometric designs, Z plasties or W plasties to camouflage the straight line visible scaring. The use of long-lasting resorbable sutures was tried too. Recently, the use of botulinum toxin during revision improved the results. Fractional CO2 lasers, microfat grafts, and platelet-rich plasma were added to the armamentarium. The scar is least visible if placed in the junction between the facial subunits. The aim of this study is to investigate the use of the subunit principle to improve the results of scar revision. Four patients were included in this study. Tissue expansion of the intact part of the subunit allowed shifting the scar to the junction between the affected subunit and the adjacent one. Tissue expansion, delivery of the expanders, and advancement of the flaps were successful in all patients. The fact that this is a 2-stage procedure and sacrifices some of the intact skin from the affected facial subunit, makes this technique reserved to patients with ugly facial scars who are ambitious to improve their appearance.

Intracellular APP Domain Regulates Serine-Palmitoyl-CoA Transferase Expression and Is Affected in Alzheimer's Disease

PubMed Central

Grimm, Marcus O. W.; Grösgen, Sven; Rothhaar, Tatjana L.; Burg, Verena K.; Hundsdörfer, Benjamin; Haupenthal, Viola J.; Friess, Petra; Müller, Ulrike; Fassbender, Klaus; Riemenschneider, Matthias; Grimm, Heike S.; Hartmann, Tobias

2011-01-01

Lipids play an important role as risk or protective factors in Alzheimer's disease (AD), a disease biochemically characterized by the accumulation of amyloid beta peptides (Aβ), released by proteolytic processing of the amyloid precursor protein (APP). Changes in sphingolipid metabolism have been associated to the development of AD. The key enzyme in sphingolipid de novo synthesis is serine-palmitoyl-CoA transferase (SPT). In the present study we identified a new physiological function of APP in sphingolipid synthesis. The APP intracellular domain (AICD) was found to decrease the expression of the SPT subunit SPTLC2, the catalytic subunit of the SPT heterodimer, resulting in that decreased SPT activity. AICD function was dependent on Fe65 and SPTLC2 levels are increased in APP knock-in mice missing a functional AICD domain. SPTLC2 levels are also increased in familial and sporadic AD postmortem brains, suggesting that SPT is involved in AD pathology. PMID:21660213

SPT-CL J2040–4451: An SZ-selected galaxy cluster at x=1.478 with significant ongoing star formation

DOE Office of Scientific and Technical Information (OSTI.GOV)

Bayliss, M. B.; Ashby, M. L. N.; Ruel, J.

2014-09-18

SPT-CL J2040-4451-spectroscopically confirmed at z = 1.478-is the highest-redshift galaxy cluster yet discovered via the Sunyaev-Zel'dovich effect. SPT-CL J2040-4451 was a candidate galaxy cluster identified in the first 720 deg(2) of the South Pole Telescope Sunyaev-Zel'dovich (SPT-SZ) survey, and has been confirmed in follow-up imaging and spectroscopy. From multi-object spectroscopy with Magellan-I/Baade+ IMACS we measure spectroscopic redshifts for 15 cluster member galaxies, all of which have strong [O Pi]lambda lambda 3727 emission. SPT-CL J2040-4451 has an SZ-measured mass of M-500,(SZ) = 3.2 ± 0.8 x 10 14M(circle dot) h(-1) 70, corresponding to M-200,M- (SZ) = 5.8 ± 1.4 x 10more » 14M(circle dot) h(70-)(1.) The velocity dispersion measured entirely from blue star-forming members is sv = 1500 ± 520 km s -1. The prevalence of star-forming cluster members (galaxies with > 1.5M(circle dot) yr -1 implies that this massive, high-redshift cluster is experiencing a phase of active star formation, and supports recent results showing a marked increase in star formation occurring in galaxy clusters at z greater than or similar to 1.4. We also compute the probability of finding a cluster as rare as this in the SPT-SZ survey to be > 99%, indicating that its discovery is not in tension with the concordance Lambda CDM cosmological model.« less

Structural and functional homology between the RNAPI subunits A14/A43 and the archaeal RNAP subunits E/F

PubMed Central

Meka, Hedije; Daoust, Gregoire; Bourke Arnvig, Kristine; Werner, Finn; Brick, Peter; Onesti, Silvia

2003-01-01

In the archaeal RNA polymerase and the eukaryotic RNA polymerase II, two subunits (E/F and RPB4/RPB7, respectively) form a heterodimer that reversibly associates with the core of the enzyme. Recently it has emerged that this heterodimer also has a counterpart in the other eukaryotic RNA polymerases: in particular two subunits of RNA polymerase I (A14 and A43) display genetic and biochemical characteristics that are similar to those of the RPB4 and RPB7 subunits, despite the fact that only A43 shows some sequence homology to RPB7. We demonstrate that the sequence of A14 strongly suggests the presence of a HRDC domain, a motif that is found at the C-terminus of a number of helicases and RNases. The same motif is also seen in the structure of the F subunit, suggesting a structural link between A14 and the RPB4/C17/subunit F family, even in the absence of direct sequence homology. We show that it is possible to co-express and co-purify large amounts of the recombinant A14/A43 heterodimer, indicating a tight and specific interaction between the two subunits. To shed light on the function of the heterodimer, we performed gel mobility shift assays and showed that the A14/A43 heterodimer binds single-stranded RNA in a similar way to the archaeal E/F complex. PMID:12888498

Joint measurement of lensing-galaxy correlations using SPT and DES SV data

DOE PAGES

Baxter, E. J.

2016-07-04

We measure the correlation of galaxy lensing and cosmic microwave background lensing with a set of galaxies expected to trace the matter density field. The measurements are performed using pre-survey Dark Energy Survey (DES) Science Verification optical imaging data and millimeter-wave data from the 2500 square degree South Pole Telescope Sunyaev-Zel'dovich (SPT-SZ) survey. The two lensing-galaxy correlations are jointly fit to extract constraints on cosmological parameters, constraints on the redshift distribution of the lens galaxies, and constraints on the absolute shear calibration of DES galaxy lensing measurements. We show that an attractive feature of these fits is that they are fairly insensitive to the clustering bias of the galaxies used as matter tracers. The measurement presented in this work confirms that DES and SPT data are consistent with each other and with the currently favoredmore » $$\\Lambda$$CDM cosmological model. In conclusion, it also demonstrates that joint lensing-galaxy correlation measurement considered here contains a wealth of information that can be extracted using current and future surveys.« less

Joint measurement of lensing-galaxy correlations using SPT and DES SV data

DOE Office of Scientific and Technical Information (OSTI.GOV)

Baxter, E. J.

We measure the correlation of galaxy lensing and cosmic microwave background lensing with a set of galaxies expected to trace the matter density field. The measurements are performed using pre-survey Dark Energy Survey (DES) Science Verification optical imaging data and millimeter-wave data from the 2500 square degree South Pole Telescope Sunyaev-Zel'dovich (SPT-SZ) survey. The two lensing-galaxy correlations are jointly fit to extract constraints on cosmological parameters, constraints on the redshift distribution of the lens galaxies, and constraints on the absolute shear calibration of DES galaxy lensing measurements. We show that an attractive feature of these fits is that they are fairly insensitive to the clustering bias of the galaxies used as matter tracers. The measurement presented in this work confirms that DES and SPT data are consistent with each other and with the currently favoredmore » $$\\Lambda$$CDM cosmological model. In conclusion, it also demonstrates that joint lensing-galaxy correlation measurement considered here contains a wealth of information that can be extracted using current and future surveys.« less

Joint measurement of lensing–galaxy correlations using SPT and DES SV data

DOE Office of Scientific and Technical Information (OSTI.GOV)

Baxter, E.; Clampitt, J.; Giannantonio, T.

We measure the correlation of galaxy lensing and cosmic microwave background lensing with a set of galaxies expected to trace the matter density field. The measurements are performed using pre-survey Dark Energy Survey (DES) Science Verification optical imaging data and millimetre-wave data from the 2500 sq. deg. South Pole Telescope Sunyaev–Zel'dovich (SPT-SZ) survey. The two lensing–galaxy correlations are jointly fit to extract constraints on cosmological parameters, constraints on the redshift distribution of the lens galaxies, and constraints on the absolute shear calibration of DES galaxy-lensing measurements. We show that an attractive feature of these fits is that they are fairlymore » insensitive to the clustering bias of the galaxies used as matter tracers. The measurement presented in this work confirms that DES and SPT data are consistent with each other and with the currently favoured Λ cold dark matter cosmological model. It also demonstrates that joint lensing–galaxy correlation measurement considered here contains a wealth of information that can be extracted using current and future surveys.« less

SPT-CL J2040–4451: An SZ-selected galaxy cluster at z = 1.478 with significant ongoing star formation

DOE Office of Scientific and Technical Information (OSTI.GOV)

Bayliss, M. B.; Ruel, J.; Ashby, M. L. N.

2014-10-10

SPT-CL J2040–4451—spectroscopically confirmed at z = 1.478—is the highest-redshift galaxy cluster yet discovered via the Sunyaev-Zel'dovich effect. SPT-CL J2040–4451 was a candidate galaxy cluster identified in the first 720 deg{sup 2} of the South Pole Telescope Sunyaev-Zel'dovich (SPT-SZ) survey, and has been confirmed in follow-up imaging and spectroscopy. From multi-object spectroscopy with Magellan-I/Baade+IMACS we measure spectroscopic redshifts for 15 cluster member galaxies, all of which have strong [O II] λλ3727 emission. SPT-CL J2040–4451 has an SZ-measured mass of M {sub 500,} {sub SZ} = 3.2 ± 0.8 × 10{sup 14} M {sub ☉} h {sub 70}{sup −1}, corresponding to Mmore » {sub 200,} {sub SZ} = 5.8 ± 1.4 × 10{sup 14} M {sub ☉} h {sub 70}{sup −1}. The velocity dispersion measured entirely from blue star-forming members is σ {sub v} = 1500 ± 520 km s{sup –1}. The prevalence of star-forming cluster members (galaxies with >1.5 M {sub ☉} yr{sup –1}) implies that this massive, high-redshift cluster is experiencing a phase of active star formation, and supports recent results showing a marked increase in star formation occurring in galaxy clusters at z ≳ 1.4. We also compute the probability of finding a cluster as rare as this in the SPT-SZ survey to be >99%, indicating that its discovery is not in tension with the concordance ΛCDM cosmological model.« less

A MEASUREMENT OF SECONDARY COSMIC MICROWAVE BACKGROUND ANISOTROPIES FROM THE 2500 SQUARE-DEGREE SPT-SZ SURVEY

DOE Office of Scientific and Technical Information (OSTI.GOV)

George, E. M.; Reichardt, C. L.; Aird, K. A.

2015-01-28

We present measurements of secondary cosmic microwave background (CMB) anisotropies and cosmic infrared background (CIB) fluctuations using data from the South Pole Telescope (SPT) covering the complete 2540 deg(2) SPT-SZ survey area. Data in the three SPT-SZ frequency bands centered at 95, 150, and 220 GHz, are used to produce six angular power spectra (three single-frequency auto-spectra and three cross-spectra) covering the multipole range 2000 < ℓ < 11, 000 (angular scales 5' gsim θ gsim 1'). These are the most precise measurements of the angular power spectra at ℓ > 2500 at these frequencies. The main contributors to the power spectra at these angular scales and frequencies are the primary CMB, CIB, thermal and kinematic Sunyaev-Zel'dovich effects (tSZ and kSZ), and radio galaxies. We include a constraint on the tSZ power from a measurement of the tSZ bispectrum from 800 deg(2) of the SPT-SZ survey. We measure the tSZ power at 143  GHz to bemore » $$D^{\\rm tSZ}_{3000} = 4.08^{+0.58}_{-0.67}\\,\\mu {\\rm K}^2{}$$ and the kSZ power to be $$D^{\\rm kSZ}_{3000} = 2.9 \\pm 1.3\\, \\mu {\\rm K}^2{}$$. The data prefer positive kSZ power at 98.1% CL. We measure a correlation coefficient of $$\\xi = 0.113^{+0.057}_{-0.054}$$ between sources of tSZ and CIB power, with ξ < 0 disfavored at a confidence level of 99.0%. The constraint on kSZ power can be interpreted as an upper limit on the duration of reionization. When the post-reionization homogeneous kSZ signal is accounted for, we find an upper limit on the duration Δz < 5.4  at 95% CL.« less

A measurement of secondary cosmic microwave background anisotropies from the 2500 square-degree SPT-SZ survey

DOE PAGES

George, E. M.; Reichardt, C. L.; Aird, K. A.; ...

2015-01-28

Here, we present measurements of secondary cosmic microwave background (CMB) anisotropies and cosmic infrared background (CIB) fluctuations using data from the South Pole Telescope (SPT) covering the complete 2540 deg2 SPT-SZ survey area. Data in the three SPT-SZ frequency bands centered at 95, 150, and 220 GHz, are used to produce six angular power spectra (three single-frequency auto-spectra and three cross-spectra) covering the multipole range 2000 < ℓ < 11, 000 (angular scales 5' gsim θ gsim 1'). These are the most precise measurements of the angular power spectra at ℓ > 2500 at these frequencies. The main contributors to the power spectra at these angular scales and frequencies are the primary CMB, CIB, thermal and kinematic Sunyaev-Zel'dovich effects (tSZ and kSZ), and radio galaxies. We include a constraint on the tSZ power from a measurement of the tSZ bispectrum from 800 deg2 of the SPT-SZ survey. We measure the tSZ power at 143 GHz to bemore » $$D^{\\rm tSZ}_{3000} = 4.08^{+0.58}_{-0.67}\\,\\mu {\\rm K}^2{}$$ and the kSZ power to be $$D^{\\rm kSZ}_{3000} = 2.9 \\pm 1.3\\, \\mu {\\rm K}^2{}$$. The data prefer positive kSZ power at 98.1% CL. We measure a correlation coefficient of $$\\xi = 0.113^{+0.057}_{-0.054}$$ between sources of tSZ and CIB power, with ξ < 0 disfavored at a confidence level of 99.0%. The constraint on kSZ power can be interpreted as an upper limit on the duration of reionization. When the post-reionization homogeneous kSZ signal is accounted for, we find an upper limit on the duration Δz < 5.4 at 95% CL.« less

Performance Evaluation of the SPT-140

NASA Technical Reports Server (NTRS)

Manzella, David; Sarmiento, Charles; Sankovic, John; Haag, Tom

1997-01-01

As part of an on-going cooperative program with industry, an engineering model SPT-140 Hall thruster, which may be suitable for orbit insertion and station-keeping of geosynchronous communication satellites, was evaluated with respect to thrust and radiated electromagnetic interference at the NASA Lewis Research Center. Performance measurements were made using a laboratory model propellant feed system and commercial power supplies. The engine was operated in a space simulation chamber capable of providing background pressures of 4 x 10(exp -6) Torr or less during thruster operation. Thrust was measured at input powers ranging from 1.5 to 5 kilowatts with two different output filter configurations. The broadband electromagnetic emission spectra generated by the engine was also measured for a range of frequencies from 0.01 to 18,000 Mhz. These results are compared to the noise threshold of the measurement system and MIL-STD-461C where appropriate.

Cosmological constraints from galaxy clusters in the 2500 square-degree SPT-SZ survey

DOE PAGES

Haan, T. de; Benson, B. A.; Bleem, L. E.; ...

2016-11-18

Here, we present cosmological parameter constraints obtained from galaxy clusters identified by their Sunyaev–Zel’dovich effect signature in the 2500 square-degree South Pole Telescope Sunyaev Zel’dovich (SPT-SZ) survey. We consider the 377 cluster candidates identified atmore » $$z\\gt 0.25$$ with a detection significance greater than five, corresponding to the 95% purity threshold for the survey. We compute constraints on cosmological models using the measured cluster abundance as a function of mass and redshift. We include additional constraints from multi-wavelength observations, including Chandra X-ray data for 82 clusters and a weak lensing-based prior on the normalization of the mass-observable scaling relations. Assuming a spatially flat ΛCDM cosmology, we combine the cluster data with a prior on H (0) and find $${\\sigma }_{8}=0.784\\pm 0.039$$ and $${{\\rm{\\Omega }}}_{m}=0.289\\pm 0.042$$, with the parameter combination $${\\sigma }_{8}{({{\\rm{\\Omega }}}_{m}/0.27)}^{0.3}=0.797\\pm 0.031$$. These results are in good agreement with constraints from the cosmic microwave background (CMB) from SPT, WMAP, and Planck, as well as with constraints from other cluster data sets. We also consider several extensions to ΛCDM, including models in which the equation of state of dark energy w, the species-summed neutrino mass, and/or the effective number of relativistic species ($${N}_{\\mathrm{eff}}$$) are free parameters. When combined with constraints from the Planck CMB, H (0), baryon acoustic oscillation, and SNe, adding the SPT cluster data improves the w constraint by 14%, to $$w=-1.023\\pm 0.042$$.« less

Cosmological constraints from galaxy clusters in the 2500 square-degree SPT-SZ survey

DOE Office of Scientific and Technical Information (OSTI.GOV)

Haan, T. de; Benson, B. A.; Bleem, L. E.

Here, we present cosmological parameter constraints obtained from galaxy clusters identified by their Sunyaev–Zel’dovich effect signature in the 2500 square-degree South Pole Telescope Sunyaev Zel’dovich (SPT-SZ) survey. We consider the 377 cluster candidates identified atmore » $$z\\gt 0.25$$ with a detection significance greater than five, corresponding to the 95% purity threshold for the survey. We compute constraints on cosmological models using the measured cluster abundance as a function of mass and redshift. We include additional constraints from multi-wavelength observations, including Chandra X-ray data for 82 clusters and a weak lensing-based prior on the normalization of the mass-observable scaling relations. Assuming a spatially flat ΛCDM cosmology, we combine the cluster data with a prior on H (0) and find $${\\sigma }_{8}=0.784\\pm 0.039$$ and $${{\\rm{\\Omega }}}_{m}=0.289\\pm 0.042$$, with the parameter combination $${\\sigma }_{8}{({{\\rm{\\Omega }}}_{m}/0.27)}^{0.3}=0.797\\pm 0.031$$. These results are in good agreement with constraints from the cosmic microwave background (CMB) from SPT, WMAP, and Planck, as well as with constraints from other cluster data sets. We also consider several extensions to ΛCDM, including models in which the equation of state of dark energy w, the species-summed neutrino mass, and/or the effective number of relativistic species ($${N}_{\\mathrm{eff}}$$) are free parameters. When combined with constraints from the Planck CMB, H (0), baryon acoustic oscillation, and SNe, adding the SPT cluster data improves the w constraint by 14%, to $$w=-1.023\\pm 0.042$$.« less

Cross-correlation of gravitational lensing from DES Science Verification data with SPT and Planck lensing

DOE Office of Scientific and Technical Information (OSTI.GOV)

Kirk, D.; Omori, Y.; Benoit-Lévy, A.

We measure the cross-correlation between weak lensing of galaxy images and of the cosmic microwave background (CMB). The effects of gravitational lensing on different sources will be correlated if the lensing is caused by the same mass fluctuations. We use galaxy shape measurements from 139 degmore » $$^{2}$$ of the Dark Energy Survey (DES) Science Verification data and overlapping CMB lensing from the South Pole Telescope (SPT) and Planck. The DES source galaxies have a median redshift of $$z_{\\rm med} {\\sim} 0.7$$, while the CMB lensing kernel is broad and peaks at $$z{\\sim}2$$. The resulting cross-correlation is maximally sensitive to mass fluctuations at $$z{\\sim}0.44$$. Assuming the Planck 2015 best-fit cosmology, the amplitude of the DES$$\\times$$SPT cross-power is found to be $$A = 0.88 \\pm 0.30$$ and that from DES$$\\times$$Planck to be $$A = 0.86 \\pm 0.39$$, where $A=1$ corresponds to the theoretical prediction. These are consistent with the expected signal and correspond to significances of $$2.9 \\sigma$$ and $$2.2 \\sigma$$ respectively. We demonstrate that our results are robust to a number of important systematic effects including the shear measurement method, estimator choice, photometric redshift uncertainty and CMB lensing systematics. Significant intrinsic alignment of galaxy shapes would increase the cross-correlation signal inferred from the data; we calculate a value of $$A = 1.08 \\pm 0.36$$ for DES$$\\times$$SPT when we correct the observations with a simple IA model. With three measurements of this cross-correlation now existing in the literature, there is not yet reliable evidence for any deviation from the expected LCDM level of cross-correlation, given the size of the statistical uncertainties and the significant impact of systematic errors, particularly IAs. We provide forecasts for the expected signal-to-noise of the combination of the five-year DES survey and SPT-3G.« less

Simplified Numerical Description of SPT Operations

NASA Technical Reports Server (NTRS)

Manzella, David H.

1995-01-01

A simplified numerical model of the plasma discharge within the SPT-100 stationary plasma thruster was developed to aid in understanding thruster operation. A one dimensional description was used. Non-axial velocities were neglected except for the azimuthal electron velocity. A nominal operating condition of 4.5 mg/s of xenon anode flow was considered with 4.5 Amperes of discharge current, and a peak radial magnetic field strength of 130 Gauss. For these conditions, the calculated results indicated ionization fractions of 0.99 near the thruster exit with a potential drop across the discharge of approximately 250 Volts. Peak calculated electron temperatures were found to be sensitive to the choice of total ionization cross section for ionization of atomic xenon by electron bombardment and ranged from 51 eV to 60 eV. The calculated ionization fraction, potential drop, and electron number density agree favorably with previous experiments. Calculated electron temperatures are higher than previously measured.

The GPI transamidase complex subunit PbGPI16 of Plasmodium berghei is important for inducing experimental cerebral malaria.

PubMed

Liu, Qingyang; Zhao, Yan; Zheng, Li; Zhu, Xiaotong; Cui, Liwang; Cao, Yaming

2018-05-21

In animal models of experimental cerebral malaria (ECM), the glycosylphosphatidylinositols (GPIs) and GPI anchors are the major factors that induce nuclear factor kappa B (NF-κB) activation and proinflammatory responses, which contribute to malaria pathogenesis. GPIs and GPI anchors are transported to the cell surface via a process called GPI transamidation, which involves the GPI transamidase (GPI-T) complex. In this study, we showed that GPI16, one of the GPI-T subunits, is highly conserved among Plasmodium species. Genetic knockout of pbgpi16 ( Δpbgpi16 ) in the rodent malaria parasite Plasmodium berghei ANKA strain led to a significant reduction of the amount of GPIs in the membranes of merozoites as well as surface display of several GPI-anchored merozoite surface proteins. Compared with the wild-type parasites, Δpbgpi16 parasites in C57BL/6 mice caused much less NF-κB activation and elicited a substantially attenuated T helper type 1 response. As a result, Δpbgpi16 -infected mice displayed much less severe brain pathology and considerably fewer Δpbgpi16 -infected mice died from ECM. This study corroborated the GPI toxin as a significant inducer of ECM and further suggested that vaccines against parasite GPIs may be a promising strategy to limit the severity of malaria. Copyright © 2018 American Society for Microbiology.

A MEASUREMENT OF SECONDARY COSMIC MICROWAVE BACKGROUND ANISOTROPIES FROM THE 2500 SQUARE-DEGREE SPT-SZ SURVEY

DOE Office of Scientific and Technical Information (OSTI.GOV)

George, E. M.; Reichardt, C. L.; Harrington, N. L.

2015-02-01

We present measurements of secondary cosmic microwave background (CMB) anisotropies and cosmic infrared background (CIB) fluctuations using data from the South Pole Telescope (SPT) covering the complete 2540 deg{sup 2} SPT-SZ survey area. Data in the three SPT-SZ frequency bands centered at 95, 150, and 220 GHz, are used to produce six angular power spectra (three single-frequency auto-spectra and three cross-spectra) covering the multipole range 2000 < ℓ < 11, 000 (angular scales 5' ≳ θ ≳ 1'). These are the most precise measurements of the angular power spectra at ℓ > 2500 at these frequencies. The main contributors to the powermore » spectra at these angular scales and frequencies are the primary CMB, CIB, thermal and kinematic Sunyaev-Zel'dovich effects (tSZ and kSZ), and radio galaxies. We include a constraint on the tSZ power from a measurement of the tSZ bispectrum from 800 deg{sup 2} of the SPT-SZ survey. We measure the tSZ power at 143  GHz to be D{sub 3000}{sup tSZ}=4.08{sub −0.67}{sup +0.58} μK{sup 2} and the kSZ power to be D{sub 3000}{sup kSZ}=2.9±1.3 μK{sup 2}. The data prefer positive kSZ power at 98.1% CL. We measure a correlation coefficient of ξ=0.113{sub −0.054}{sup +0.057} between sources of tSZ and CIB power, with ξ < 0 disfavored at a confidence level of 99.0%. The constraint on kSZ power can be interpreted as an upper limit on the duration of reionization. When the post-reionization homogeneous kSZ signal is accounted for, we find an upper limit on the duration Δz < 5.4  at 95% CL.« less

16 CFR 5.63 - Evidence; transcript; in camera orders; proposed findings of fact and conclusions of law.

Code of Federal Regulations, 2012 CFR

2012-01-01

... 16 Commercial Practices 1 2012-01-01 2012-01-01 false Evidence; transcript; in camera orders; proposed findings of fact and conclusions of law. 5.63 Section 5.63 Commercial Practices FEDERAL TRADE... Concerning Postemployment Conflict of Interest § 5.63 Evidence; transcript; in camera orders; proposed...

16 CFR 5.63 - Evidence; transcript; in camera orders; proposed findings of fact and conclusions of law.

Code of Federal Regulations, 2014 CFR

2014-01-01

... 16 Commercial Practices 1 2014-01-01 2014-01-01 false Evidence; transcript; in camera orders; proposed findings of fact and conclusions of law. 5.63 Section 5.63 Commercial Practices FEDERAL TRADE... Concerning Postemployment Conflict of Interest § 5.63 Evidence; transcript; in camera orders; proposed...

16 CFR 5.63 - Evidence; transcript; in camera orders; proposed findings of fact and conclusions of law.

Code of Federal Regulations, 2013 CFR

2013-01-01

... 16 Commercial Practices 1 2013-01-01 2013-01-01 false Evidence; transcript; in camera orders; proposed findings of fact and conclusions of law. 5.63 Section 5.63 Commercial Practices FEDERAL TRADE... Concerning Postemployment Conflict of Interest § 5.63 Evidence; transcript; in camera orders; proposed...

16 CFR 5.63 - Evidence; transcript; in camera orders; proposed findings of fact and conclusions of law.

Code of Federal Regulations, 2011 CFR

2011-01-01

... 16 Commercial Practices 1 2011-01-01 2011-01-01 false Evidence; transcript; in camera orders; proposed findings of fact and conclusions of law. 5.63 Section 5.63 Commercial Practices FEDERAL TRADE... Concerning Postemployment Conflict of Interest § 5.63 Evidence; transcript; in camera orders; proposed...

Utah State Office of Education Fingertip Facts, 2015-16

ERIC Educational Resources Information Center

Utah State Office of Education, 2016

2016-01-01

Fingertip Facts is a compendium of some of the most frequently requested data sets from the Utah State Office of Education. This year's Fingertip Facts includes the following data sets: SAGE Testing, 2014-15; 2015 Public Education General Fund; 2014-15 Public School Enrollment Demographics; Public Schools by Grade Level, 2014-15; Number of…

VizieR Online Data Catalog: SPT-SZ survey galaxy clusters optical spectroscopy (Ruel+, 2014)

NASA Astrophysics Data System (ADS)

Ruel, J.; Bazin, G.; Bayliss, M.; Brodwin, M.; Foley, R. J.; Stalder, B.; Aird, K. A.; Armstrong, R.; Ashby, M. L. N.; Bautz, M.; Benson, B. A.; Bleem, L. E.; Bocquet, S.; Carlstrom, J. E.; Chang, C. L.; Chapman, S. C.; Cho, H. M.; Clocchiatti, A.; Crawford, T. M.; Crites, A. T.; de Haan, T.; Desai, S.; Dobbs, M. A.; Dudley, J. P.; Forman, W. R.; George, E. M.; Gladders, M. D.; Gonzalez, A. H.; Halverson, N. W.; Harrington, N. L.; High, F. W.; Holder, G. P.; Holzapfel, W. L.; Hrubes, J. D.; Jones, C.; Joy, M.; Keisler, R.; Knox, L.; Lee, A. T.; Leitch, E. M.; Liu, J.; Lueker, M.; Luong-van, D.; Mantz, A.; Marrone, D. P.; McDonald, M.; McMahon, J. J.; Mehl, J.; Meyer, S. S.; Mocanu, L.; Mohr, J. J.; Montroy, T. E.; Murray, S. S.; Natoli, T.; Nurgaliev, D.; Padin, S.; Plagge, T.; Pryke, C.; Reichardt, C. L.; Rest, A.; Ruhl, J. E.; Saliwanchik, B. R.; Saro, A.; Sayre, J. T.; Schaffer, K. K.; Shaw, L.; Shirokoff, E.; Song, J.; Suhada, R.; Spieler, H. G.; Stanford, S. A.; Staniszewski, Z.; Starsk, A. A.; Story, K.; Stubbs, C. W.; van Engelen, A.; Vanderlinde, K.; Vieira, J. D.; Vikhlinin, A.; Williamson, R.; Zahn, O.; Zenteno, A.

2017-04-01

Most of the galaxy clusters for which we report spectroscopic observations were published as SPT cluster detections (and new discoveries) in Vanderlinde et al. (2010ApJ...722.1180V), Williamson et al. (2011ApJ...738..139W), and Reichardt et al. (2013, J/ApJ/763/127); we refer the reader to those publications for details of the SPT observations. The spectroscopic observations presented in this work are the first of our ongoing follow-up program. The data were taken from 2008 to 2012 using the Gemini Multi Object Spectrograph (GMOS; Hook et al. 2004PASP..116..425H) on Gemini South, the Focal Reducer and low dispersion Spectrograph (FORS2; Appenzeller et al. 1998Msngr..94....1A) on VLT Antu, the Inamori Magellan Areal Camera and Spectrograph (IMACS; Dressler et al. 2006SPIE.6269E..0FD) on Magellan Baade, and the Low Dispersion Survey Spectrograph (LDSS339; Allington-Smith et al. 1994PASP..106..983A) on Magellan Clay. (2 data files).

Bright monomeric photoactivatable red fluorescent protein for two-color super-resolution sptPALM of live cells.

PubMed

Subach, Fedor V; Patterson, George H; Renz, Malte; Lippincott-Schwartz, Jennifer; Verkhusha, Vladislav V

2010-05-12

Rapidly emerging techniques of super-resolution single-molecule microscopy of living cells rely on the continued development of genetically encoded photoactivatable fluorescent proteins. On the basis of monomeric TagRFP, we have developed a photoactivatable TagRFP protein that is initially dark but becomes red fluorescent after violet light irradiation. Compared to other monomeric dark-to-red photoactivatable proteins including PAmCherry, PATagRFP has substantially higher molecular brightness, better pH stability, substantially less sensitivity to blue light, and better photostability in both ensemble and single-molecule modes. Spectroscopic analysis suggests that PATagRFP photoactivation is a two-step photochemical process involving sequential one-photon absorbance by two distinct chromophore forms. True monomeric behavior, absence of green fluorescence, and single-molecule performance in live cells make PATagRFP an excellent protein tag for two-color imaging techniques, including conventional diffraction-limited photoactivation microscopy, super-resolution photoactivated localization microscopy (PALM), and single particle tracking PALM (sptPALM) of living cells. Two-color sptPALM imaging was demonstrated using several PATagRFP tagged transmembrane proteins together with PAGFP-tagged clathrin light chain. Analysis of the resulting sptPALM images revealed that single-molecule transmembrane proteins, which are internalized into a cell via endocytosis, colocalize in space and time with plasma membrane domains enriched in clathrin light-chain molecules.

Variation in GABA-A subunit gene copy number in an autistic patient with mosaic 4 p duplication (p12p16).

PubMed

Kakinuma, Hiroaki; Ozaki, Mamoru; Sato, Hitoshi; Takahashi, Hiroaki

2008-09-05

Autism has been associated with chromosomal aberrations, including duplications at chromosome 4, and the identification of genetic factors contributing to the etiology of this disease is the focus of much research. Here we report a Japanese girl with mosaic of chromosome 4p duplication, mos 46,XX,dup(4)(p12p16)[54]/46,XX[6], who was diagnosed with autism at 3 years of age. Fluorescence in situ hybridization (FISH) with probes covering the region spanning a cluster of the gamma aminobutyric acid A (GABA-A) receptor subunit genes in the proximal short arm of chromosome 4 demonstrated total three signals for the GABRG1, GABRA4, and GABRA2 genes, but only two signals for GABRB1. This suggests that aberrant copy number of the GABA-A receptor subunit genes may contribute to the etiology of autism in this patient. 2007 Wiley-Liss, Inc.

Na+, K+-ATPase β1 subunit associates with α1 subunit modulating a "higher-NKA-in-hyposmotic media" response in gills of euryhaline milkfish, Chanos chanos.

PubMed

Hu, Yau-Chung; Chu, Keng-Fu; Yang, Wen-Kai; Lee, Tsung-Han

2017-10-01

The euryhaline milkfish (Chanos chanos) is a popular aquaculture species that can be cultured in fresh water, brackish water, or seawater in Southeast Asia. In gills of the milkfish, Na + , K + -ATPase (i.e., NKA; sodium pump) responds to salinity challenges including changes in mRNA abundance, protein amount, and activity. The functional pump is composed of a heterodimeric protein complex composed of α- and β-subunits. Among the NKA genes, α1-β1 isozyme comprises the major form of NKA subunits in mammalian osmoregulatory organs; however, most studies on fish gills have focused on the α1 subunit and did not verify the α1-β1 isozyme. Based on the sequenced milkfish transcriptome, an NKA β1 subunit gene was identified that had the highest amino acid homology to β233, a NKA β1 subunit paralog originally identified in the eel. Despite this high level of homology to β233, phylogenetic analysis and the fact that only a single NKA β1 subunit gene exists in the milkfish suggest that the milkfish gene should be referred to as the NKA β1 subunit gene. The results of accurate domain prediction of the β1 subunit, co-localization of α1 and β1 subunits in epithelial ionocytes, and co-immunoprecipitation of α1 and β1 subunits, indicated the formation of a α1-β1 complex in milkfish gills. Moreover, when transferred to hyposmotic media (fresh water) from seawater, parallel increases in branchial mRNA and protein expression of NKA α1 and β1 subunits suggested their roles in hypo-osmoregulation of euryhaline milkfish. This study molecularly characterized the NKA β1 subunit and provided the first evidence for an NKA α1-β1 association in gill ionocytes of euryhaline teleosts.

The p40 Subunit of Interleukin (IL)-12 Promotes Stabilization and Export of the p35 Subunit

PubMed Central

Jalah, Rashmi; Rosati, Margherita; Ganneru, Brunda; Pilkington, Guy R.; Valentin, Antonio; Kulkarni, Viraj; Bergamaschi, Cristina; Chowdhury, Bhabadeb; Zhang, Gen-Mu; Beach, Rachel Kelly; Alicea, Candido; Broderick, Kate E.; Sardesai, Niranjan Y.; Pavlakis, George N.; Felber, Barbara K.

2013-01-01

IL-12 is a 70-kDa heterodimeric cytokine composed of the p35 and p40 subunits. To maximize cytokine production from plasmid DNA, molecular steps controlling IL-12p70 biosynthesis at the posttranscriptional and posttranslational levels were investigated. We show that the combination of RNA/codon-optimized gene sequences and fine-tuning of the relative expression levels of the two subunits within a cell resulted in increased production of the IL-12p70 heterodimer. We found that the p40 subunit plays a critical role in enhancing the stability, intracellular trafficking, and export of the p35 subunit. This posttranslational regulation mediated by the p40 subunit is conserved in mammals. Based on these findings, dual gene expression vectors were generated, producing an optimal ratio of the two subunits, resulting in a ∼1 log increase in human, rhesus, and murine IL-12p70 production compared with vectors expressing the wild type sequences. Such optimized DNA plasmids also produced significantly higher levels of systemic bioactive IL-12 upon in vivo DNA delivery in mice compared with plasmids expressing the wild type sequences. A single therapeutic injection of an optimized murine IL-12 DNA plasmid showed significantly more potent control of tumor development in the B16 melanoma cancer model in mice. Therefore, the improved IL-12p70 DNA vectors have promising potential for in vivo use as molecular vaccine adjuvants and in cancer immunotherapy. PMID:23297419

Skin test hypersensitivity for childhood asthma in Istanbul during a period of 16 years.

PubMed

Akcakaya, N; Cokugras, H; Camcioglu, Y; Ozdemir, M

2005-01-01

For diagnosis of allergic disorders, besides history and physical examination, many in vivo and in vitro laboratory tests are used. Skin prick test (SPT) is an easily performed and valuable test in children. Our aim was to evaluate the SPT results of asthmatic children according to age and gender, in Istanbul and its neighborhood, followed for a period of 16 years. Consecutive 5080 asthmatic children, aged 1 to 18 years, admitted to Pediatric Allergy Policlinics of Istanbul University Cerrahpasa Medicine Faculty from 1987 to 2003, were screened retrospectively for 10 frequently exposed allergens. 3086 cases (61 %) had SPT reactivity for one and/or for multiple allergens; the SPT reactivity percentages of the exposed allergens was 50 % (2554 cases) for house dust mites DP, 49 % (2462 cases) for house dust mites DF, 15 % (784 cases) for cat dander, 10 % (525 cases) for dog dander, 4 % (228 cases) for lamb wool, 10 % (504 cases) for wheat, 3 % (162 cases) for egg white, 6 % (345 cases) for hazelnut pollen, 6 % (318 cases) for Candida Albicans, 6 % (326 cases) for Aspergillus Fumigatus. While house dust mites were determined as the predominant allergen for each age groups in this study, allergy against cat dander was the third important allergen in sequence, as cat is a domestic pet which is frequently kept at home in Turkey.

The augmentation of heat transfer in a pipe flow using a swirling perforated twisted (SPT) tape insert

NASA Astrophysics Data System (ADS)

Ahmad, Shahrokh; Oishe, Sadia Noon; Rahman, Md. Lutfor

2017-12-01

The purpose of this research work is to increase the heat transfer coefficient by operating the heat exchangers at smaller revolution per minute. This signifies an achievement of reduction of pressure drop corresponding to less operating cost. This study has used two types of SPT tape insert to observe the various heat transfer coefficient, heat transfer rate and heat transfer augmentation efficiency. One tape was fully twisted and another tape was partially twisted. The shape of the SPT tape creates turbulence effect. The turbulence flow (swirl flow) generated by SPT tape promotes greater mixing and high heat transfer coefficients. An arrangement scheme has been developed for the experimental investigation. For remarking the rate of change of heat transfer, temperature has been measured numerically through the temperature sensors with various flow rates and RPM. The volume flow rate was varied from 10.3448276 LPM to 21.045574 LPM and the rotation of the perforated twisted tape was varied from 50 RPM to 400 RPM. Finally the research study demonstrates the effectiveness of the results of the proposed approaches. It is observed that the suggested method of heat transfer augmentations is much more effective than existing methods, since it results in an increase in heat transfer area and also an increase in the heat transfer coefficient and reduction of cost in the industrial sectors.

An ALMA view of the interstellar medium of the z = 4.77 lensed starburst SPT-S J213242-5802.9

NASA Astrophysics Data System (ADS)

Béthermin, M.; De Breuck, C.; Gullberg, B.; Aravena, M.; Bothwell, M. S.; Chapman, S. C.; Gonzalez, A. H.; Greve, T. R.; Litke, K.; Ma, J.; Malkan, M.; Marrone, D. P.; Murphy, E. J.; Spilker, J. S.; Stark, A. A.; Strandet, M.; Vieira, J. D.; Weiß, A.; Welikala, N.

2016-02-01

We present ALMA detections of the [NII] 205 μm and CO(12-11) emission lines, and the tentative detection of [CI] - for the strongly lensed (μ = 5.7 ± 0.5) dusty, star-forming galaxy SPT-S J213242-5802.9 (hereafter SPT2132-58) at z = 4.77. The [NII] and CO(12-11) lines are detected at 11.5 and 8.5σ levels, respectively, by our band 6 observations. The [CI] line is detected at 3.2σ after a reanalysis of existing band 3 data. The [CI] luminosity implies a gas mass of (3.8 ± 1.2) × 1010M⊙, and, consequently, a very short depletion timescale of 34 ± 13 Myr and a CO luminosity to gas mass conversion factor αCO of 1.0 ± 0.3 M⊙ (K km s-1 pc2)-1. SPT2132-58 is an extreme starburst with an intrinsic star formation rate of 1100 ± 200 M⊙/yr. We find a [CII]/[NII] ratio of 26 ± 6, which is the highest ratio reported at z > 4. This suggests that SPT2132-58 hosts an evolved interstellar medium (0.5 Z⊙< Z < 1.5 Z⊙), which may be dominated by photodissociation regions. The CO(2-1) and CO(5-4) transitions have lower CO to far-infrared ratios than local and high-redshift samples, while CO(12-11) is similar to these samples, suggesting the presence of an additional very excited component or an active galactic nucleus.

Mass calibration and cosmological analysis of the SPT-SZ galaxy cluster sample using velocity dispersion σ v and x-ray Y X measurements

DOE PAGES

Bocquet, S.; Saro, A.; Mohr, J. J.; ...

2015-01-30

Here, we present a velocity-dispersion-based mass calibration of the South Pole Telescope Sunyaev-Zel'dovich effect survey (SPT-SZ) galaxy cluster sample. Using a homogeneously selected sample of 100 cluster candidates from 720 deg 2 of the survey along with 63 velocity dispersion (σ v) and 16 X-ray Y X measurements of sample clusters, we simultaneously calibrate the mass-observable relation and constrain cosmological parameters. Our method accounts for cluster selection, cosmological sensitivity, and uncertainties in the mass calibrators. The calibrations using σ v and Y X are consistent at the 0.6σ level, with the σ v calibration preferring ~16% higher masses. We usemore » the full SPTCL data set (SZ clusters+σ v+Y X) to measure σ 8(Ωm/0.27) 0.3 = 0.809 ± 0.036 within a flat ΛCDM model. The SPT cluster abundance is lower than preferred by either the WMAP9 or Planck+WMAP9 polarization (WP) data, but assuming that the sum of the neutrino masses is m ν = 0.06 eV, we find the data sets to be consistent at the 1.0σ level for WMAP9 and 1.5σ for Planck+WP. Allowing for larger Σm ν further reconciles the results. When we combine the SPTCL and Planck+WP data sets with information from baryon acoustic oscillations and Type Ia supernovae, the preferred cluster masses are 1.9σ higher than the Y X calibration and 0.8σ higher than the σ v calibration. Given the scale of these shifts (~44% and ~23% in mass, respectively), we execute a goodness-of-fit test; it reveals no tension, indicating that the best-fit model provides an adequate description of the data. Using the multi-probe data set, we measure Ω m = 0.299 ± 0.009 and σ8 = 0.829 ± 0.011. Within a νCDM model we find Σm ν = 0.148 ± 0.081 eV. We present a consistency test of the cosmic growth rate using SPT clusters. Allowing both the growth index γ and the dark energy equation-of-state parameter w to vary, we find γ = 0.73 ± 0.28 and w = –1.007 ± 0.065, demonstrating that the eΣxpansion and

Mass Calibration and Cosmological Analysis of the SPT-SZ Galaxy Cluster Sample Using Velocity Dispersion σ v and X-Ray Y X Measurements

NASA Astrophysics Data System (ADS)

Bocquet, S.; Saro, A.; Mohr, J. J.; Aird, K. A.; Ashby, M. L. N.; Bautz, M.; Bayliss, M.; Bazin, G.; Benson, B. A.; Bleem, L. E.; Brodwin, M.; Carlstrom, J. E.; Chang, C. L.; Chiu, I.; Cho, H. M.; Clocchiatti, A.; Crawford, T. M.; Crites, A. T.; Desai, S.; de Haan, T.; Dietrich, J. P.; Dobbs, M. A.; Foley, R. J.; Forman, W. R.; Gangkofner, D.; George, E. M.; Gladders, M. D.; Gonzalez, A. H.; Halverson, N. W.; Hennig, C.; Hlavacek-Larrondo, J.; Holder, G. P.; Holzapfel, W. L.; Hrubes, J. D.; Jones, C.; Keisler, R.; Knox, L.; Lee, A. T.; Leitch, E. M.; Liu, J.; Lueker, M.; Luong-Van, D.; Marrone, D. P.; McDonald, M.; McMahon, J. J.; Meyer, S. S.; Mocanu, L.; Murray, S. S.; Padin, S.; Pryke, C.; Reichardt, C. L.; Rest, A.; Ruel, J.; Ruhl, J. E.; Saliwanchik, B. R.; Sayre, J. T.; Schaffer, K. K.; Shirokoff, E.; Spieler, H. G.; Stalder, B.; Stanford, S. A.; Staniszewski, Z.; Stark, A. A.; Story, K.; Stubbs, C. W.; Vanderlinde, K.; Vieira, J. D.; Vikhlinin, A.; Williamson, R.; Zahn, O.; Zenteno, A.

2015-02-01

We present a velocity-dispersion-based mass calibration of the South Pole Telescope Sunyaev-Zel'dovich effect survey (SPT-SZ) galaxy cluster sample. Using a homogeneously selected sample of 100 cluster candidates from 720 deg2 of the survey along with 63 velocity dispersion (σ v ) and 16 X-ray Y X measurements of sample clusters, we simultaneously calibrate the mass-observable relation and constrain cosmological parameters. Our method accounts for cluster selection, cosmological sensitivity, and uncertainties in the mass calibrators. The calibrations using σ v and Y X are consistent at the 0.6σ level, with the σ v calibration preferring ~16% higher masses. We use the full SPTCL data set (SZ clusters+σ v +Y X) to measure σ8(Ωm/0.27)0.3 = 0.809 ± 0.036 within a flat ΛCDM model. The SPT cluster abundance is lower than preferred by either the WMAP9 or Planck+WMAP9 polarization (WP) data, but assuming that the sum of the neutrino masses is ∑m ν = 0.06 eV, we find the data sets to be consistent at the 1.0σ level for WMAP9 and 1.5σ for Planck+WP. Allowing for larger ∑m ν further reconciles the results. When we combine the SPTCL and Planck+WP data sets with information from baryon acoustic oscillations and Type Ia supernovae, the preferred cluster masses are 1.9σ higher than the Y X calibration and 0.8σ higher than the σ v calibration. Given the scale of these shifts (~44% and ~23% in mass, respectively), we execute a goodness-of-fit test; it reveals no tension, indicating that the best-fit model provides an adequate description of the data. Using the multi-probe data set, we measure Ωm = 0.299 ± 0.009 and σ8 = 0.829 ± 0.011. Within a νCDM model we find ∑m ν = 0.148 ± 0.081 eV. We present a consistency test of the cosmic growth rate using SPT clusters. Allowing both the growth index γ and the dark energy equation-of-state parameter w to vary, we find γ = 0.73 ± 0.28 and w = -1.007 ± 0.065, demonstrating that the expansion and the growth

PRDM16 enhances nuclear receptor-dependent transcription of the brown fat-specific Ucp1 gene through interactions with Mediator subunit MED1.

PubMed

Iida, Satoshi; Chen, Wei; Nakadai, Tomoyoshi; Ohkuma, Yoshiaki; Roeder, Robert G

2015-02-01

PR domain-containing 16 (PRDM16) induces expression of brown fat-specific genes in brown and beige adipocytes, although the underlying transcription-related mechanisms remain largely unknown. Here, in vitro studies show that PRDM16, through its zinc finger domains, directly interacts with the MED1 subunit of the Mediator complex, is recruited to the enhancer of the brown fat-specific uncoupling protein 1 (Ucp1) gene through this interaction, and enhances thyroid hormone receptor (TR)-driven transcription in a biochemically defined system in a Mediator-dependent manner, thus providing a direct link to the general transcription machinery. Complementary cell-based studies show that upon forskolin treatment, PRDM16 induces Ucp1 expression in undifferentiated murine embryonic fibroblasts, that this induction depends on MED1 and TR, and, consistent with a direct effect, that PRDM16 is recruited to the Ucp1 enhancer. Related studies have defined MED1 and PRDM16 interaction domains important for Ucp1 versus Ppargc1a induction by PRDM16. These results reveal novel mechanisms for PRDM16 function through the Mediator complex. © 2015 Iida et al.; Published by Cold Spring Harbor Laboratory Press.

ALMA OBSERVATIONS OF SPT-DISCOVERED, STRONGLY LENSED, DUSTY, STAR-FORMING GALAXIES

DOE Office of Scientific and Technical Information (OSTI.GOV)

Hezaveh, Y. D.; Marrone, D. P.; Spilker, J. S.

2013-04-20

We present Atacama Large Millimeter/submillimeter Array (ALMA) 860 {mu}m imaging of four high-redshift (z = 2.8-5.7) dusty sources that were detected using the South Pole Telescope (SPT) at 1.4 mm and are not seen in existing radio to far-infrared catalogs. At 1.''5 resolution, the ALMA data reveal multiple images of each submillimeter source, separated by 1''-3'', consistent with strong lensing by intervening galaxies visible in near-IR imaging of these sources. We describe a gravitational lens modeling procedure that operates on the measured visibilities and incorporates self-calibration-like antenna phase corrections as part of the model optimization, which we use to interpretmore » the source structure. Lens models indicate that SPT0346-52, located at z = 5.7, is one of the most luminous and intensely star-forming sources in the universe with a lensing corrected FIR luminosity of 3.7 Multiplication-Sign 10{sup 13} L{sub Sun} and star formation surface density of 4200 M{sub Sun} yr{sup -1} kpc{sup -2}. We find magnification factors of 5 to 22, with lens Einstein radii of 1.''1-2.''0 and Einstein enclosed masses of 1.6-7.2 Multiplication-Sign 10{sup 11} M{sub Sun }. These observations confirm the lensing origin of these objects, allow us to measure their intrinsic sizes and luminosities, and demonstrate the important role that ALMA will play in the interpretation of lensed submillimeter sources.« less

16 CFR 460.14 - How retailers must handle fact sheets.

Code of Federal Regulations, 2010 CFR

2010-01-01

....14 Section 460.14 Commercial Practices FEDERAL TRADE COMMISSION TRADE REGULATION RULES LABELING AND... are likely to notice them. For example, you can put them in a display, and let customers take copies... customers where the fact sheets are. You need not make the fact sheets available to customers if you display...

Sunyaev-Zel'dovich Effect and X-ray Scaling Relations from Weak-Lensing Mass Calibration of 32 SPT Selected Galaxy Clusters

DOE Office of Scientific and Technical Information (OSTI.GOV)

Dietrich, J.P.; et al.

Uncertainty in the mass-observable scaling relations is currently the limiting factor for galaxy cluster based cosmology. Weak gravitational lensing can provide a direct mass calibration and reduce the mass uncertainty. We present new ground-based weak lensing observations of 19 South Pole Telescope (SPT) selected clusters and combine them with previously reported space-based observations of 13 galaxy clusters to constrain the cluster mass scaling relations with the Sunyaev-Zel'dovich effect (SZE), the cluster gas massmore » $$M_\\mathrm{gas}$$, and $$Y_\\mathrm{X}$$, the product of $$M_\\mathrm{gas}$$ and X-ray temperature. We extend a previously used framework for the analysis of scaling relations and cosmological constraints obtained from SPT-selected clusters to make use of weak lensing information. We introduce a new approach to estimate the effective average redshift distribution of background galaxies and quantify a number of systematic errors affecting the weak lensing modelling. These errors include a calibration of the bias incurred by fitting a Navarro-Frenk-White profile to the reduced shear using $N$-body simulations. We blind the analysis to avoid confirmation bias. We are able to limit the systematic uncertainties to 6.4% in cluster mass (68% confidence). Our constraints on the mass-X-ray observable scaling relations parameters are consistent with those obtained by earlier studies, and our constraints for the mass-SZE scaling relation are consistent with the the simulation-based prior used in the most recent SPT-SZ cosmology analysis. We can now replace the external mass calibration priors used in previous SPT-SZ cosmology studies with a direct, internal calibration obtained on the same clusters.« less

Limits on the fluctuating part of y-type distortion monopole from Planck and SPT results

NASA Astrophysics Data System (ADS)

Khatri, Rishi; Sunyaev, Rashid

2015-08-01

We use the published Planck and SPT cluster catalogs [1,2] and recently published y-distortion maps [3] to put strong observational limits on the contribution of the fluctuating part of the y-type distortions to the y-distortion monopole. Our bounds are 5.4× 10-8 < langle yrangle < 2.2× 10-6. Our upper bound is a factor of 6.8 stronger than the currently best upper 95% confidence limit from COBE-FIRAS of langle yrangle <15× 10-6. In the standard cosmology, large scale structure is the only source of such distortions and our limits therefore constrain the baryonic physics involved in the formation of the large scale structure. Our lower limit, from the detected clusters in the Planck and SPT catalogs, also implies that a Pixie-like experiment should detect the y-distortion monopole at >27-σ. The biggest sources of uncertainty in our upper limit are the monopole offsets between different HFI channel maps that we estimate to be <10-6.

Temperature-dependent subunit exchange and chaperone-like activities of Hsp16.3, a small heat shock protein from Mycobacterium tuberculosis.

PubMed

Fu, Xinmiao; Chang, Zengyi

2004-04-02

Small heat shock proteins (sHsps) usually exist as oligomers that undergo dynamic oligomeric dissociation/re-association, with the dissociated oligomers as active forms to bind substrate proteins under heat shock conditions. In this study, however, we found that Hsp16.3, one sHsp from Mycobacterium tuberculosis, is able to sensitively modulate its chaperone-like activity in a range of physiological temperatures (from 25 to 37.5 degrees C) while its native oligomeric size is still maintained. Further analysis demonstrated that Hsp16.3 exposes higher hydrophobic surfaces upon temperatures increasing and that a large soluble complex between Hsp16.3 and substrate is formed only in the condition of heating temperature up to 35 and 37.5 degrees C. Structural analysis by fluorescence anisotropy showed that Hsp16.3 nonameric structure becomes more dynamic and variable at elevated temperatures. Moreover, subunit exchange between Hsp16.3 oligomers was found to occur faster upon temperatures increasing as revealed by fluorescence energy resonance transfer. These observations indicate that Hsp16.3 is able to modulate its chaperone activity by adjusting the dynamics of oligomeric dissociation/re-association process while maintaining its static oligomeric size unchangeable. A kinetic model is therefore proposed to explain the mechanism of sHsps-binding substrate proteins through oligomeric dissociation. The present study also implied that Hsp16.3 is at least capable of binding non-native proteins in vivo while expressing in the host organism that survives at 37 degrees C.

A 205 Hour Krypton Propellant Life Test of the SPT-100 Operating at 3 kW

DTIC Science & Technology

2013-09-01

advantageous, such as orbit raising missions. Bismuth’s main drawback is that the metal must be vaporized to be ionized and accelerated within a Hall...the performance of the SPT-100 operating on krypton was characterized using an inverted pendulum thrust stand over a wide range of thruster operating

Integration Tests of the 4 kW-Class High Voltage Hall Accelerator Power Processing Unit with the HiVHAc and the SPT-140 Hall Effect Thrusters

NASA Technical Reports Server (NTRS)

Kamhawi, Hani; Pinero, Luis; Haag, Thomas; Huang, Wensheng; Ahern, Drew; Liang, Ray; Shilo, Vlad

2016-01-01

NASA's Science Mission Directorate is sponsoring the development of a 4 kW-class Hall propulsion system for implementation in NASA science and exploration missions. The main components of the system include the High Voltage Hall Accelerator (HiVHAc), an engineering model power processing unit (PPU) developed by Colorado Power Electronics, and a xenon flow control module (XFCM) developed by VACCO Industries. NASA Glenn Research Center is performing integrated tests of the Hall thruster propulsion system. This paper presents results from integrated tests of the PPU and XFCM with the HiVHAc engineering development thruster and a SPT-140 thruster provided by Space System Loral. The results presented in this paper demonstrate thruster discharge initiation along with open-loop and closed-loop control of the discharge current with anode flow for both the HiVHAc and the SPT-140 thrusters. Integrated tests with the SPT-140 thruster indicated that the PPU was able to repeatedly initiate the thruster's discharge, achieve steady state operation, and successfully throttle the thruster between 1.5 and 4.5 kW. The measured SPT-140 performance was identical to levels reported by Space Systems Loral.

Integration Tests of the 4 kW-class High Voltage Hall Accelerator Power Processing Unit with the HiVHAc and the SPT-140 Hall Effect Thrusters

NASA Technical Reports Server (NTRS)

Kamhawi, Hani; Pinero, Luis; Haag, Thomas; Huang, Wensheng; Ahern, Drew; Liang, Ray; Shilo, Vlad

2016-01-01

NASAs Science Mission Directorate is sponsoring the development of a 4 kW-class Hall propulsion system for implementation in NASA science and exploration missions. The main components of the system include the High Voltage Hall Accelerator (HiVHAc), an engineering model power processing unit (PPU) developed by Colorado Power Electronics, and a xenon flow control module (XFCM) developed by VACCO Industries. NASA Glenn Research Center is performing integrated tests of the Hall thruster propulsion system. This presentation presents results from integrated tests of the PPU and XFCM with the HiVHAc engineering development thruster and a SPT-140 thruster provided by Space System Loral. The results presented in this paper demonstrate thruster discharge initiation, open-loop and closed-loop control of the discharge current with anode flow for both the HiVHAc and the SPT-140 thrusters. Integrated tests with the SPT-140 thruster indicated that the PPU was able to repeatedly initiate the thrusters discharge, achieve steady state operation, and successfully throttle the thruster between 1.5 and 4.5 kW. The measured SPT-140 performance was identical to levels reported by Space Systems Loral.

Cigarette smoking during pregnancy regulates the expression of specific nicotinic acetylcholine receptor (nAChR) subunits in the human placenta

DOE Office of Scientific and Technical Information (OSTI.GOV)

Machaalani, R., E-mail: rita.machaalani@sydney.edu.au; Bosch Institute, The University of Sydney, NSW 2006; The Children's Hospital at Westmead, NSW 2145

Smoking during pregnancy is associated with low birth weight, premature delivery, and neonatal morbidity and mortality. Nicotine, a major pathogenic compound of cigarette smoke, binds to the nicotinic acetylcholine receptors (nAChRs). A total of 16 nAChR subunits have been identified in mammals (9 α, 4 β, and 1 δ, γ and ε subunits). The effect of cigarette smoking on the expression of these subunits in the placenta has not yet been determined, thus constituting the aim of this study. Using RT-qPCR and western blotting, this study investigated all 16 mammalian nAChR subunits in the normal healthy human placenta, and comparedmore » mRNA and protein expressions in the placentas from smokers (n = 8) to controls (n = 8). Our data show that all 16 subunit mRNAs are expressed in the normal, non-diseased human placenta and that the expression of α2, α3, α4, α9, β2 and β4 subunits is greater than the other subunits. For mRNA, cigarette smoke exposure was associated with increased expression of the α9 subunit, and decreased expression of the δ subunit. At the protein level, expression of both α9 and δ was increased. Thus, cigarette smoking in pregnancy is sufficient to regulate nAChR subunits in the placenta, specifically α9 and δ subunits, and could contribute to the adverse effects of vasoconstriction and decreased re-epithelialisation (α9), and increased calcification and apoptosis (δ), seen in the placentas of smoking women. - Highlights: • All 16 mammalian nAChR subunits are expressed in the human placenta. • Cigarette smoking increases α9 mRNA and protein in the placenta. • Cigarette smoking decreases δ mRNA but increases δ protein in the placenta.« less

Plasma plume diagnostics of low power stationary plasma thruster (SPT-20M8) with collisional radiative model

NASA Astrophysics Data System (ADS)

Uttamsing Rajput, Rajendrasing; Alona, Khaustova; Loyan, Andriy V.

2017-03-01

Electric propulsion offers higher specific impulse compared to the chemical propulsion systems. It reduces the overall propellant mass and enables high operational lifetimes. Scientific Technological Center of Space Power and Energy (STC SPE), KhAI is involved in designing, manufacturing and testing of stationary plasma thrusters (SPT). Efforts are made to perform plasma diagnostics with corona and collisional radiative models (C-R model), as expected corona model falls short below 4 eV because of the heavy particle collisions elimination, whereas the C-R model's applicability is confirmed. Several tests are performed to analyze the electron temperature at various operational parameters of thruster like discharge voltage and mass flow rate. SPT-20M8 far and near-field plumes diagnostics are performed. Feasibility of C-R model by comparing its result to optical emission spectroscopy (OES) to investigate the electron temperature is validated with the probe measurements within the 10% of discrepancy.

Limits on the fluctuating part of y-type distortion monopole from Planck and SPT results

DOE Office of Scientific and Technical Information (OSTI.GOV)

Khatri, Rishi; Sunyaev, Rashid, E-mail: khatri@mpa-garching.mpg.de, E-mail: sunyaev@mpa-garching.mpg.de

2015-08-01

We use the published Planck and SPT cluster catalogs [1,2] and recently published y-distortion maps [3] to put strong observational limits on the contribution of the fluctuating part of the y-type distortions to the y-distortion monopole. Our bounds are 5.4× 10{sup −8} < ( y) < 2.2× 10{sup −6}. Our upper bound is a factor of 6.8 stronger than the currently best upper 95% confidence limit from COBE-FIRAS of ( y) <15× 10{sup −6}. In the standard cosmology, large scale structure is the only source of such distortions and our limits therefore constrain the baryonic physics involved in the formation of the large scale structure. Our lower limit, from themore » detected clusters in the Planck and SPT catalogs, also implies that a Pixie-like experiment should detect the y-distortion monopole at >27-σ. The biggest sources of uncertainty in our upper limit are the monopole offsets between different HFI channel maps that we estimate to be <10{sup −6}.« less

Significant expression of thyroid transcription factor-1 in pulmonary squamous cell carcinoma detected by SPT24 monoclonal antibody and CSA-II system.

PubMed

Kashima, Kenji; Hashimoto, Hisashi; Nishida, Haruto; Arakane, Motoki; Yada, Naomi; Daa, Tsutomu; Yokoyama, Shigeo

2014-01-01

In contrast to the usefulness of thyroid transcription factor-1 (TTF-1) in distinguishing primary adenocarcinoma of the lung from metastatic lesions, TTF-1 expression in pulmonary squamous cell carcinoma is reported to be at low level and not a suitable immunohistochemical marker. We hypothesized that the highly sensitive detection system, CSA-II, can visualize even faint expression of TTF-1 in pulmonary squamous cell carcinoma. In this study, 2 commercially available clones of TTF-1 monoclonal antibody, 8G7G3/1 and SPT24, were used for staining 38 cases of pulmonary squamous cell carcinoma, in combination with the CSA-II and the conventional detection system, EnVision. The combined use of the 8G7G3/1 clone with EnVision and CSA-II showed a positive reaction in only 1 and 4 cases, respectively. The use of SPT24 clone showed positive staining in 5 cases with EnVision and in 20 of 38 cases (52.6%) with the CSA-II. Interestingly, positive staining by the SPT24-CSA-II technique of samples from tissue blocks preserved for <2 years was 73.6% compared with only 31.5% in those preserved for >2 years. In addition, a 6-month preservation of the cut sections resulted in stain fading and decreased positivity (50%), compared with freshly cut sections. We conclude that the use of the SPT24 monoclonal antibody with the CSA-II system can detect even weak expression of TTF-1 in pulmonary squamous cell carcinoma. This staining technique can potentially allow the discrimination of primary squamous cell carcinoma of the lung from metastatic lesions, especially in freshly prepared paraffin sections.

Comparison of SPT and V s-based liquefaction analyses: a case study in Erciş (Van, Turkey)

NASA Astrophysics Data System (ADS)

Akkaya, İsmail; Özvan, Ali; Akin, Mutluhan; Akin, Müge K.; Övün, Uğur

2017-12-01

Liquefaction which is one of the most destructive ground deformations occurs during an earthquake in saturated or partially saturated silty and sandy soils, which may cause serious damages such as settlement and tilting of structures due to shear strength loss of soils. Standard (SPT) and cone (CPT) penetration tests as well as the shear wave velocity (V s)-based methods are commonly used for the determination of liquefaction potential. In this research, it was aimed to compare the SPT and V s-based liquefaction analysis methods by generating different earthquake scenarios. Accordingly, the Erciş residential area, which was mostly affected by the 2011 Van earthquake (M w = 7.1), was chosen as the model site. Erciş (Van, Turkey) and its surroundings settle on an alluvial plain which consists of silty and sandy layers with shallow groundwater level. Moreover, Çaldıran, Erciş-Kocapınar and Van Fault Zones are the major seismic sources of the region which have a significant potential of producing large magnitude earthquakes. After liquefaction assessments, the liquefaction potential in the western part of the region and in the coastal regions nearby the Lake Van is found to be higher than the other locations. Thus, it can be stated that the soil tightness and groundwater level dominantly control the liquefaction potential. In addition, the lateral spreading and sand boiling spots observed after the 23rd October 2011 Van earthquake overlap the scenario boundaries predicted in this study. Eventually, the use of V s-based liquefaction analysis in collaboration with the SPT results is quite advantageous to assess the rate of liquefaction in a specific area.

Comparison of SPT and V s-based liquefaction analyses: a case study in Erciş (Van, Turkey)

NASA Astrophysics Data System (ADS)

Akkaya, İsmail; Özvan, Ali; Akin, Mutluhan; Akin, Müge K.; Övün, Uğur

2018-02-01

Liquefaction which is one of the most destructive ground deformations occurs during an earthquake in saturated or partially saturated silty and sandy soils, which may cause serious damages such as settlement and tilting of structures due to shear strength loss of soils. Standard (SPT) and cone (CPT) penetration tests as well as the shear wave velocity ( V s)-based methods are commonly used for the determination of liquefaction potential. In this research, it was aimed to compare the SPT and V s-based liquefaction analysis methods by generating different earthquake scenarios. Accordingly, the Erciş residential area, which was mostly affected by the 2011 Van earthquake ( M w = 7.1), was chosen as the model site. Erciş (Van, Turkey) and its surroundings settle on an alluvial plain which consists of silty and sandy layers with shallow groundwater level. Moreover, Çaldıran, Erciş-Kocapınar and Van Fault Zones are the major seismic sources of the region which have a significant potential of producing large magnitude earthquakes. After liquefaction assessments, the liquefaction potential in the western part of the region and in the coastal regions nearby the Lake Van is found to be higher than the other locations. Thus, it can be stated that the soil tightness and groundwater level dominantly control the liquefaction potential. In addition, the lateral spreading and sand boiling spots observed after the 23rd October 2011 Van earthquake overlap the scenario boundaries predicted in this study. Eventually, the use of V s-based liquefaction analysis in collaboration with the SPT results is quite advantageous to assess the rate of liquefaction in a specific area.

GALAXY CLUSTERS DISCOVERED VIA THE SUNYAEV-ZEL'DOVICH EFFECT IN THE 2500-SQUARE-DEGREE SPT-SZ SURVEY

DOE Office of Scientific and Technical Information (OSTI.GOV)

Bleem, L. E.; Stalder, B.; de Haan, T.

2015-01-29

We present a catalog of galaxy clusters selected via their Sunyaev-Zel'dovich (SZ) effect signature from 2500 deg(2) of South Pole Telescope (SPT) data. This work represents the complete sample of clusters detected at high significance in the 2500 deg(2) SPT-SZ survey, which was completed in 2011. A total of 677 (409) cluster candidates are identified above a signal-to-noise threshold of ξ = 4.5 (5.0). Ground- and space-based optical and near-infrared (NIR) imaging confirms overdensities of similarly colored galaxies in the direction of 516 (or 76%) of the ξ > 4.5 candidates and 387 (or 95%) of the ξ > 5 candidates, the measured purity is consistent with expectations from simulations. Of these confirmed clusters, 415 were first identified in SPT data, including 251 new discoveries reported in this work. We estimate photometric redshifts for all candidates with identified optical and/or NIR counterparts, we additionally report redshifts derived from spectroscopic observations for 141 of these systems. The mass threshold of the catalog is roughly independent of redshift above z ~ 0.25 leading to a sample of massive clusters that extends to high redshift. The median mass of the sample is M (500c)(ρ(crit))more » $$\\sim 3.5\\times 10^{14}\\,M_\\odot \\,h_{70}^{-1}$$, the median redshift is z (med) = 0.55, and the highest-redshift systems are at z > 1.4. The combination of large redshift extent, clean selection, and high typical mass makes this cluster sample of particular interest for cosmological analyses and studies of cluster formation and evolution.« less

GALAXY CLUSTERS DISCOVERED VIA THE SUNYAEV-ZEL'DOVICH EFFECT IN THE 2500-SQUARE-DEGREE SPT-SZ SURVEY

DOE Office of Scientific and Technical Information (OSTI.GOV)

Bleem, L. E.; Stalder, B.; de Haan, T.

2015-01-29

We present a catalog of galaxy clusters selected via their Sunyaev-Zel'dovich (SZ) effect signature from 2500 deg(2) of South Pole Telescope (SPT) data. This work represents the complete sample of clusters detected at high significance in the 2500 deg(2) SPT-SZ survey, which was completed in 2011. A total of 677 (409) cluster candidates are identified above a signal-to-noise threshold of xi = 4.5 (5.0). Ground-and space-based optical and near-infrared (NIR) imaging confirms overdensities of similarly colored galaxies in the direction of 516 (or 76%) of the xi > 4.5 candidates and 387 (or 95%) of the xi > 5 candidates;more » the measured purity is consistent with expectations from simulations. Of these confirmed clusters, 415 were first identified in SPT data, including 251 new discoveries reported in this work. We estimate photometric redshifts for all candidates with identified optical and/or NIR counterparts; we additionally report redshifts derived from spectroscopic observations for 141 of these systems. The mass threshold of the catalog is roughly independent of redshift above z similar to 0.25 leading to a sample of massive clusters that extends to high redshift. The median mass of the sample is M-500c(rho(crit)) similar to 3.5 x 10(14) M-circle dot h(70)(-1) 70, the median redshift is z(med) = 0.55, and the highest-redshift systems are at z > 1.4. The combination of large redshift extent, clean selection, and high typical mass makes this cluster sample of particular interest for cosmological analyses and studies of cluster formation and evolution.« less

Sandia Simple Particle Tracking (Sandia SPT) v. 1.0

DOE Office of Scientific and Technical Information (OSTI.GOV)

Anthony, Stephen M.

2015-06-15

Sandia SPT is designed as software to accompany a book chapter being published a methods chapter which provides an introduction on how to label and track individual proteins. The Sandia Simple Particle Tracking code uses techniques common to the image processing community, where its value is that it facilitates implementing the methods described in the book chapter by providing the necessary open-source code. The code performs single particle spot detection (or segmentation and localization) followed by tracking (or connecting the detected particles into trajectories). The book chapter, which along with the headers in each file, constitutes the documentation for themore » code is: Anthony, S.M.; Carroll-Portillo, A.; Timlon, J.A., Dynamics and Interactions of Individual Proteins in the Membrane of Living Cells. In Anup K. Singh (Ed.) Single Cell Protein Analysis Methods in Molecular Biology. Springer« less

Identification of critical residues of subunit H in its interaction with subunit E of the A-ATP synthase from Methanocaldococcus jannaschii.

PubMed

Gayen, Shovanlal; Balakrishna, Asha M; Biuković, Goran; Yulei, Wu; Hunke, Cornelia; Grüber, Gerhard

2008-04-01

The boomerang-like H subunit of A(1)A(0) ATP synthase forms one of the peripheral stalks connecting the A(1) and A(0) sections. Structural analyses of the N-terminal part (H1-47) of subunit H of the A(1)A(0) ATP synthase from Methanocaldococcus jannaschii have been performed by NMR spectroscopy. Our initial NMR structural calculations for H1-47 indicate that amino acid residues 7-44 fold into a single alpha-helical structure. Using the purified N- (E1-100) and C-terminal domains (E101-206) of subunit E, NMR titration experiments revealed that the N-terminal residues Met1-6, Lys10, Glu11, Ala15, Val20 and Glu24 of H1-47 interact specifically with the N-terminal domain E1-100 of subunit E. A more detailed picture regarding the residues of E1-100 involved in this association was obtained by titration studies using the N-terminal peptides E1-20, E21-40 and E41-60. These data indicate that the N-terminal tail E41-60 interacts with the N-terminal amino acids of H1-47, and this has been confirmed by fluorescence correlation spectroscopy results. Analysis of (1)H-(15)N heteronuclear single quantum coherence (HSQC) spectra of the central stalk subunit F in the presence and absence of E101-206 show no obvious interaction between the C-terminal domain of E and subunit F. The data presented provide, for the first time, structural insights into the interaction of subunits E and H, and their arrangement within A(1)A(0) ATP synthase.

Subunit interactions in horse spleen apoferritin. Dissociation by extremes of pH

PubMed Central

Crichton, Robert R.; Bryce, Charles F. A.

1973-01-01

1. The dissociation of horse spleen apoferritin as a function of pH was analysed by sedimentation-velocity techniques. The oligomer is stable in the range pH2.8–10.6. Between pH2.8 and 1.6 and 10.6 and 13.0 both oligomer and subunits can be detected. At pH values between 1.6 and 1.0 the subunit is the only species observed, although below pH1.0 aggregation of the subunits to a particle sedimenting much faster than the oligomer occurs. 2. When apoferritin is first dissociated into subunits at low pH values and then dialysed into buffers of pH1.5–5.0, the subunit reassociates to oligomer in the pH range 3.1–4.3. 3. U.v.-difference spectroscopy was used to study conformational changes occurring during the dissociation process. The difference spectrum in acid can be accounted for by the transfer of four to five tyrosine residues/subunit from the interior of the protein into the solvent. This process is reversed on reassociation, but shows the same hysteresis as found by sedimentation techniques. The difference spectrum in alkali is more complex, but is consistent with the deprotonation of tyrosine residues, which appear to have rather high pK values. 4. In addition to the involvement of tyrosine residues in the conformational change at low pH values, spectral evidence is presented that one tryptophan residue/subunit also changes its environment before dissociation and subsequent to reassociation. 5. Analysis of the dissociation and reassociation of apoferritin at low pH values suggests that this is a co-operative process involving protonation and deprotonation of at least two carboxyl functions of rather low intrinsic pK. The dissociation at alkaline pH values does not appear to be co-operative. 6. Of the five tyrosine residues/subunit only one can be nitrated with tetranitromethane. Guanidination of lysine residues results in the modification of seven out of a total of nine residues/subunit. Nine out of the ten arginine residues/subunit react with

A Performance and Plume Comparison of Xenon and Krypton Propellant on the SPT-100

DTIC Science & Technology

2012-07-02

HET (1.35 kW), performance measurements were made using an inverted pendulum thrust stand. The plume was also characterized by a Faraday probe and RPA...performance reduction for the case of the flight model SPT-100 HET (1.35 kW), per- formance measurements were made using an inverted pendulum thrust stand...where high thrust at reduced specific impulse is advantageous, such as orbit raising missions. Bismuth’s main drawback is that the metal must be

GALAXY CLUSTERS DISCOVERED VIA THE SUNYAEV-ZEL'DOVICH EFFECT IN THE 2500-SQUARE-DEGREE SPT-SZ SURVEY

DOE Office of Scientific and Technical Information (OSTI.GOV)

Bleem, L. E.; Carlstrom, J. E.; Chang, C. L.

2015-02-01

We present a catalog of galaxy clusters selected via their Sunyaev-Zel'dovich (SZ) effect signature from 2500 deg{sup 2} of South Pole Telescope (SPT) data. This work represents the complete sample of clusters detected at high significance in the 2500 deg{sup 2} SPT-SZ survey, which was completed in 2011. A total of 677 (409) cluster candidates are identified above a signal-to-noise threshold of ξ = 4.5 (5.0). Ground- and space-based optical and near-infrared (NIR) imaging confirms overdensities of similarly colored galaxies in the direction of 516 (or 76%) of the ξ > 4.5 candidates and 387 (or 95%) of the ξ > 5more » candidates; the measured purity is consistent with expectations from simulations. Of these confirmed clusters, 415 were first identified in SPT data, including 251 new discoveries reported in this work. We estimate photometric redshifts for all candidates with identified optical and/or NIR counterparts; we additionally report redshifts derived from spectroscopic observations for 141 of these systems. The mass threshold of the catalog is roughly independent of redshift above z ∼ 0.25 leading to a sample of massive clusters that extends to high redshift. The median mass of the sample is M {sub 500c}(ρ{sub crit}) ∼3.5×10{sup 14} M{sub ⊙} h{sub 70}{sup −1}, the median redshift is z {sub med} = 0.55, and the highest-redshift systems are at z > 1.4. The combination of large redshift extent, clean selection, and high typical mass makes this cluster sample of particular interest for cosmological analyses and studies of cluster formation and evolution.« less

Pharmacological consequences of the coexpression of BK channel α and auxiliary β subunits

PubMed Central

Torres, Yolima P.; Granados, Sara T.; Latorre, Ramón

2014-01-01

Coded by a single gene (Slo1, KCM) and activated by depolarizing potentials and by a rise in intracellular Ca2+ concentration, the large conductance voltage- and Ca2+-activated K+ channel (BK) is unique among the superfamily of K+ channels. BK channels are tetramers characterized by a pore-forming α subunit containing seven transmembrane segments (instead of the six found in voltage-dependent K+ channels) and a large C terminus composed of two regulators of K+ conductance domains (RCK domains), where the Ca2+-binding sites reside. BK channels can be associated with accessory β subunits and, although different BK modulatory mechanisms have been described, greater interest has recently been placed on the role that the β subunits may play in the modulation of BK channel gating due to its physiological importance. Four β subunits have currently been identified (i.e., β1, β2, β3, and β4) and despite the fact that they all share the same topology, it has been shown that every β subunit has a specific tissue distribution and that they modify channel kinetics as well as their pharmacological properties and the apparent Ca2+ sensitivity of the α subunit in different ways. Additionally, different studies have shown that natural, endogenous, and synthetic compounds can modulate BK channels through β subunits. Considering the importance of these channels in different pathological conditions, such as hypertension and neurological disorders, this review focuses on the mechanisms by which these compounds modulate the biophysical properties of BK channels through the regulation of β subunits, as well as their potential therapeutic uses for diseases such as those mentioned above. PMID:25346693

Pharmacological consequences of the coexpression of BK channel α and auxiliary β subunits.

PubMed

Torres, Yolima P; Granados, Sara T; Latorre, Ramón

2014-01-01

Coded by a single gene (Slo1, KCM) and activated by depolarizing potentials and by a rise in intracellular Ca(2+) concentration, the large conductance voltage- and Ca(2+)-activated K(+) channel (BK) is unique among the superfamily of K(+) channels. BK channels are tetramers characterized by a pore-forming α subunit containing seven transmembrane segments (instead of the six found in voltage-dependent K(+) channels) and a large C terminus composed of two regulators of K(+) conductance domains (RCK domains), where the Ca(2+)-binding sites reside. BK channels can be associated with accessory β subunits and, although different BK modulatory mechanisms have been described, greater interest has recently been placed on the role that the β subunits may play in the modulation of BK channel gating due to its physiological importance. Four β subunits have currently been identified (i.e., β1, β2, β3, and β4) and despite the fact that they all share the same topology, it has been shown that every β subunit has a specific tissue distribution and that they modify channel kinetics as well as their pharmacological properties and the apparent Ca(2+) sensitivity of the α subunit in different ways. Additionally, different studies have shown that natural, endogenous, and synthetic compounds can modulate BK channels through β subunits. Considering the importance of these channels in different pathological conditions, such as hypertension and neurological disorders, this review focuses on the mechanisms by which these compounds modulate the biophysical properties of BK channels through the regulation of β subunits, as well as their potential therapeutic uses for diseases such as those mentioned above.

Subunit rotation of ATP synthase embedded in membranes: a or β subunit rotation relative to the c subunit ring

PubMed Central

Nishio, Kazuaki; Iwamoto-Kihara, Atsuko; Yamamoto, Akitsugu; Wada, Yoh; Futai, Masamitsu

2002-01-01

ATP synthase FoF1 (α3β3γδɛab2c10–14) couples an electrochemical proton gradient and a chemical reaction through the rotation of its subunit assembly. In this study, we engineered FoF1 to examine the rotation of the catalytic F1 β or membrane sector Fo a subunit when the Fo c subunit ring was immobilized; a biotin-tag was introduced onto the β or a subunit, and a His-tag onto the c subunit ring. Membrane fragments were obtained from Escherichia coli cells carrying the recombinant plasmid for the engineered FoF1 and were immobilized on a glass surface. An actin filament connected to the β or a subunit rotated counterclockwise on the addition of ATP, and generated essentially the same torque as one connected to the c ring of FoF1 immobilized through a His-tag linked to the α or β subunit. These results established that the γɛc10–14 and α3β3δab2 complexes are mechanical units of the membrane-embedded enzyme involved in rotational catalysis. PMID:12357031

The PA influenza virus polymerase subunit is a phosphorylated protein.

PubMed

Sanz-Ezquerro, J J; Fernández Santarén, J; Sierra, T; Aragón, T; Ortega, J; Ortín, J; Smith, G L; Nieto, A

1998-03-01

The induction of proteolysis by expression of the influenza virus PA polymerase subunit is the only biochemical activity ascribed to this protein. In the course of studying viral protein synthesis by two-dimensional gel electrophoresis, we observed the existence of several PA isoforms with different isoelectric points. These isoforms were also present when the PA gene was singly expressed in three different expression systems, indicating that a cellular activity is responsible for its post-translational modification. In vivo labelling with [32P]orthophosphate, followed by two-dimensional gel electrophoresis, clearly demonstrated the incorporation of phosphate into the PA molecule. Phosphoserine and phosphothreonine epitopes were present in PA, while phosphotyrosine residues were absent, as tested by immunoblotting with specific antibodies. These facts, as well as the presence of multiple consensus sites for casein kinase II (CKII) phosphorylation, prompted us to test the involvement of this kinase in PA covalent modification. PA protein purified by immunoprecipitation could be specifically labelled by the catalytic alpha subunit of human CKII, which was expressed and purified from bacteria. Collectively, these data demonstrate that the PA subunit of the influenza virus RNA polymerase is a phosphoprotein.

Expression of a mutated SPT15 gene in Saccharomyces cerevisiae enhances both cell growth and ethanol production in microaerobic batch, fed-batch, and simultaneous saccharification and fermentations.

PubMed

Seong, Yeong-Je; Park, Haeseong; Yang, Jungwoo; Kim, Soo-Jung; Choi, Wonja; Kim, Kyoung Heon; Park, Yong-Cheol

2017-05-01

The SPT15 gene encodes a Saccharomyces cerevisiae TATA-binding protein, which is able to globally control the transcription levels of various metabolic and regulatory genes. In this study, a SPT15 gene mutant (S42N, S78R, S163P, and I212N) was expressed in S. cerevisiae BY4741 (BSPT15-M3), of which effects on fermentative yeast properties were evaluated in a series of culture types. By applying different nitrogen sources and air supply conditions in batch culture, organic nitrogen sources and microaerobic condition were decided to be more favorable for both cell growth and ethanol production of the BSPT15-M3 strain than the control S. cerevisiae BY4741 strain expressing the SPT15 gene (BSPT15wt). Microaerobic fed-batch cultures of BSPT15-M3 with glucose shock in the presence of high ethanol content resulted in a 9.5-13.4% higher glucose consumption rate and ethanol productivity than those for the BSPT15wt strain. In addition, BSPT15-M3 showed 4.5 and 3.9% increases in ethanol productivity from cassava hydrolysates and corn starch in simultaneous saccharification and fermentation processes, respectively. It was concluded that overexpression of the mutated SPT15 gene would be a potent strategy to develop robust S. cerevisiae strains with enhanced cell growth and ethanol production abilities.

A Chandra Survey of high-redshift (0.7 < z < 0.8) clusters selected in the 100 deg^2 SPT-Pol Deep Field

NASA Astrophysics Data System (ADS)

Garmire, Gordon

2016-09-01

We propose to observe a complete sample of 10 galaxy clusters at 1e14 < M500 < 5e14 and 0.7 < z < 0.8. These systems were selected from the 100 deg^2 deep field of the SPT-Pol SZ survey. This survey are has significant complementary data, including uniform depth ATCA, Herschel, Spitzer, and DES imaging, enabling a wide variety of astrophysical and cosmological studies. This sample complements the successful SPT-XVP survey, which has a broad redshift range and a narrow mass range, by including clusters over a narrow redshift range and broad mass range. These systems are such low mass and high redshift that they will not be detected in the eRosita all-sky survey.

Protein-RNA crosslinking in Escherichia coli 30S ribosomal subunits. Identification of a 16S rRNA fragment crosslinked to protein S12 by the use of the chemical crosslinking reagent 1-ethyl-3-dimethyl-aminopropylcarbodiimide.

PubMed Central

Chiaruttini, C; Expert-Bezançon, A; Hayes, D; Ehresmann, B

1982-01-01

1-ethyl-3-dimethyl aminopropylcarbodiimide (EDC) was used to cross-link 30S ribosomal proteins to 16S rRNA within the E. coli 3OS ribosomal subunit. Covalently linked complexes containing 30S proteins and 16S rRNA, isolated by sedimentation of dissociated crosslinked 30S subunits through SDS containing sucrose gradients, were digested with RNase T1, and the resulting oligonucleotide-protein complexes were fractionated on SDS containing polyacrylamide gels. Eluted complexes containing 30S proteins S9 and S12 linked to oligonucleotides were obtained in pure form. Oligonucleotide 5'terminal labelling was successful in the case of S12 containing but not of the S9 containing complex and led to identification of the S12 bound oligonucleotide as CAACUCG which is located at positions 1316-1322 in the 16S rRNA sequence. Protein S12 is crosslinked to the terminal G of this heptanucleotide. Images PMID:6760129

THE REDSHIFT DISTRIBUTION OF DUSTY STAR-FORMING GALAXIES FROM THE SPT SURVEY

DOE Office of Scientific and Technical Information (OSTI.GOV)

Strandet, M. L.; Weiss, A.; Vieira, J. D.

2016-05-10

We use the Atacama Large Millimeter/submillimeter Array (ALMA) in Cycle 1 to determine spectroscopic redshifts of high-redshift dusty star-forming galaxies (DSFGs) selected by their 1.4 mm continuum emission in the South Pole Telescope (SPT) survey. We present ALMA 3 mm spectral scans between 84 and 114 GHz for 15 galaxies and targeted ALMA 1 mm observations for an additional eight sources. Our observations yield 30 new line detections from CO, [C i], [N ii], H{sub 2}O and NH{sub 3}. We further present Atacama Pathfinder Experiment [C ii] and CO mid- J observations for seven sources for which only a singlemore » line was detected in spectral-scan data from ALMA Cycle 0 or Cycle 1. We combine the new observations with previously published and new millimeter/submillimeter line and photometric data of the SPT-selected DSFGs to study their redshift distribution. The combined data yield 39 spectroscopic redshifts from molecular lines, a success rate of >85%. Our sample represents the largest data set of its kind today and has the highest spectroscopic completeness among all redshift surveys of high- z DSFGs. The median of the redshift distribution is z = 3.9 ± 0.4, and the highest-redshift source in our sample is at z = 5.8. We discuss how the selection of our sources affects the redshift distribution, focusing on source brightness, selection wavelength, and strong gravitational lensing. We correct for the effect of gravitational lensing and find the redshift distribution for 1.4 mm selected sources with a median redshift of z = 3.1 ± 0.3. Comparing to redshift distributions selected at shorter wavelengths from the literature, we show that selection wavelength affects the shape of the redshift distribution.« less

The Redshift Distribution of Dusty Star-forming Galaxies from the SPT Survey

NASA Astrophysics Data System (ADS)

Strandet, M. L.; Weiss, A.; Vieira, J. D.; de Breuck, C.; Aguirre, J. E.; Aravena, M.; Ashby, M. L. N.; Béthermin, M.; Bradford, C. M.; Carlstrom, J. E.; Chapman, S. C.; Crawford, T. M.; Everett, W.; Fassnacht, C. D.; Furstenau, R. M.; Gonzalez, A. H.; Greve, T. R.; Gullberg, B.; Hezaveh, Y.; Kamenetzky, J. R.; Litke, K.; Ma, J.; Malkan, M.; Marrone, D. P.; Menten, K. M.; Murphy, E. J.; Nadolski, A.; Rotermund, K. M.; Spilker, J. S.; Stark, A. A.; Welikala, N.

2016-05-01

We use the Atacama Large Millimeter/submillimeter Array (ALMA) in Cycle 1 to determine spectroscopic redshifts of high-redshift dusty star-forming galaxies (DSFGs) selected by their 1.4 mm continuum emission in the South Pole Telescope (SPT) survey. We present ALMA 3 mm spectral scans between 84 and 114 GHz for 15 galaxies and targeted ALMA 1 mm observations for an additional eight sources. Our observations yield 30 new line detections from CO, [C I], [N II], H2O and NH3. We further present Atacama Pathfinder Experiment [C II] and CO mid-J observations for seven sources for which only a single line was detected in spectral-scan data from ALMA Cycle 0 or Cycle 1. We combine the new observations with previously published and new millimeter/submillimeter line and photometric data of the SPT-selected DSFGs to study their redshift distribution. The combined data yield 39 spectroscopic redshifts from molecular lines, a success rate of >85%. Our sample represents the largest data set of its kind today and has the highest spectroscopic completeness among all redshift surveys of high-z DSFGs. The median of the redshift distribution is z = 3.9 ± 0.4, and the highest-redshift source in our sample is at z = 5.8. We discuss how the selection of our sources affects the redshift distribution, focusing on source brightness, selection wavelength, and strong gravitational lensing. We correct for the effect of gravitational lensing and find the redshift distribution for 1.4 mm selected sources with a median redshift of z = 3.1 ± 0.3. Comparing to redshift distributions selected at shorter wavelengths from the literature, we show that selection wavelength affects the shape of the redshift distribution.

The Mediator Complex Subunits MED14, MED15, and MED16 Are Involved in Defense Signaling Crosstalk in Arabidopsis.

PubMed

Wang, Chenggang; Du, Xuezhu; Mou, Zhonglin

2016-01-01

Mediator is a highly conserved protein complex that functions as a transcriptional coactivator in RNA polymerase II (RNAPII)-mediated transcription. The Arabidopsis Mediator complex has recently been implicated in plant immune responses. Here, we compared salicylic acid (SA)-, methyl jasmonate (MeJA)-, and the ethylene (ET) precursor 1-aminocyclopropane-1-carboxylic acid (ACC)-induced defense and/or wound-responsive gene expression in 14 Arabidopsis Mediator subunit mutants. Our results show that MED14, MED15, and MED16 are required for SA-activated expression of the defense marker gene PATHOEGNESIS-RELATED GENE1 , MED25 is required for MeJA-induced expression of the wound-responsive marker gene VEGATATIVE STORAGE PROTEIN1 ( VSP1 ), MED8, MED14, MED15, MED16, MED18, MED20a, MED25, MED31, and MED33A/B (MED33a and MED33B) are required for MeJA-induced expression of the defense maker gene PLANT DEFENSIN1.2 ( PDF1.2 ), and MED8, MED14, MED15, MED16, MED25, and MED33A/B are also required for ACC-triggered expression of PDF1.2 . Furthermore, we investigated the involvement of MED14, MED15, and MED16 in plant defense signaling crosstalk and found that MED14, MED15, and MED16 are required for SA- and ET-mediated suppression of MeJA-induced VSP1 expression. This result suggests that MED14, MED15, and MED16 not only relay defense signaling from the SA and JA/ET defense pathways to the RNAPII transcription machinery, but also fine-tune defense signaling crosstalk. Finally, we show that MED33A/B contributes to the necrotrophic fungal pathogen Botrytis cinerea- induced expression of the defense genes PDF1.2, HEVEIN-LIKE , and BASIC CHITINASE and is required for full-scale basal resistance to B. cinerea , demonstrating a positive role for MED33 in plant immunity against necrotrophic fungal pathogens.

A Chandra Survey of low-mass clusters at 0.8 < z < 0.9 selected in the 100 deg^2 SPT-Pol Deep Field

NASA Astrophysics Data System (ADS)

Kraft, Ralph

2016-09-01

We propose to observe a complete sample of 4 galaxy clusters at 1e14 < M500 < 3e14 and 0.8 < z < 0.9. These systems were selected from the 100 deg^2 deep field of the SPT-Pol SZ survey. This survey are has significant complementary data, including uniform depth ATCA, Herschel, Spitzer, and DES imaging, enabling a wide variety of astrophysical and cosmological studies. This sample complements the successful SPT-XVP survey, which has a broad redshift range and a narrow mass range, by including clusters over a narrow redshift range and broad mass range. These systems are such low mass and high redshift that they will not be detected in the eRosita all-sky survey.

A Chandra Survey of low-mass clusters at 0.7 < z < 0.8 selected in the 100 deg^2 SPT-Pol Deep Field

NASA Astrophysics Data System (ADS)

Kraft, Ralph

2016-09-01

We propose to observe a complete sample of 4 galaxy clusters at 1e14 < M500 < 3e14 and 0.7 < z < 0.8. These systems were selected from the 100 deg^2 deep field of the SPT-Pol SZ survey. This survey are has significant complementary data, including uniform depth ATCA, Herschel, Spitzer, and DES imaging, enabling a wide variety of astrophysical and cosmological studies. This sample complements the successful SPT-XVP survey, which has a broad redshift range and a narrow mass range, by including clusters over a narrow redshift range and broad mass range. These systems are such low mass and high redshift that they will not be detected in the eRosita all-sky survey.

FACT is a sensor of DNA torsional stress in eukaryotic cells

PubMed Central

Safina, Alfiya; Cheney, Peter; Pal, Mahadeb; Brodsky, Leonid; Ivanov, Alexander; Kirsanov, Kirill; Lesovaya, Ekaterina; Naberezhnov, Denis; Nesher, Elimelech; Koman, Igor; Wang, Dan; Wang, Jianming; Yakubovskaya, Marianna; Winkler, Duane

2017-01-01

Abstract Transitions of B-DNA to alternative DNA structures (ADS) can be triggered by negative torsional strain, which occurs during replication and transcription, and may lead to genomic instability. However, how ADS are recognized in cells is unclear. We found that the binding of candidate anticancer drug, curaxin, to cellular DNA results in uncoiling of nucleosomal DNA, accumulation of negative supercoiling and conversion of multiple regions of genomic DNA into left-handed Z-form. Histone chaperone FACT binds rapidly to the same regions via the SSRP1 subunit in curaxin-treated cells. In vitro binding of purified SSRP1 or its isolated CID domain to a methylated DNA fragment containing alternating purine/pyrimidines, which is prone to Z-DNA transition, is much stronger than to other types of DNA. We propose that FACT can recognize and bind Z-DNA or DNA in transition from a B to Z form. Binding of FACT to these genomic regions triggers a p53 response. Furthermore, FACT has been shown to bind to other types of ADS through a different structural domain, which also leads to p53 activation. Thus, we propose that FACT acts as a sensor of ADS formation in cells. Recognition of ADS by FACT followed by a p53 response may explain the role of FACT in DNA damage prevention. PMID:28082391

Prefoldin Subunits Are Protected from Ubiquitin-Proteasome System-mediated Degradation by Forming Complex with Other Constituent Subunits*

PubMed Central

Miyazawa, Makoto; Tashiro, Erika; Kitaura, Hirotake; Maita, Hiroshi; Suto, Hiroo; Iguchi-Ariga, Sanae M. M.; Ariga, Hiroyoshi

2011-01-01

The molecular chaperone prefoldin (PFD) is a complex comprised of six different subunits, PFD1-PFD6, and delivers newly synthesized unfolded proteins to cytosolic chaperonin TRiC/CCT to facilitate the folding of proteins. PFD subunits also have functions different from the function of the PFD complex. We previously identified MM-1α/PFD5 as a novel c-Myc-binding protein and found that MM-1α suppresses transformation activity of c-Myc. However, it remains unclear how cells regulate protein levels of individual subunits and what mechanisms alter the ratio of their activities between subunits and their complex. In this study, we found that knockdown of one subunit decreased protein levels of other subunits and that transfection of five subunits other than MM-1α into cells increased the level of endogenous MM-1α. We also found that treatment of cells with MG132, a proteasome inhibitor, increased the level of transfected/overexpressed MM-1α but not that of endogenous MM-1α, indicating that overexpressed MM-1α, but not endogenous MM-1α, was degraded by the ubiquitin proteasome system (UPS). Experiments using other PFD subunits showed that the UPS degraded a monomer of PFD subunits, though extents of degradation varied among subunits. Furthermore, the level of one subunit was increased after co-transfection with the respective subunit, indicating that there are specific combinations between subunits to be stabilized. These results suggest mutual regulation of protein levels among PFD subunits and show how individual subunits form the PFD complex without degradation. PMID:21478150

SB-205384 Is a Positive Allosteric Modulator of Recombinant GABAA Receptors Containing Rat α3, α5, or α6 Subunit Subtypes Coexpressed with β3 and γ2 Subunits

PubMed Central

Heidelberg, Laura S.; Warren, James W.

2013-01-01

Many drugs used to treat anxiety are positive modulators of GABAA receptors, which mediate fast inhibitory neurotransmission. The GABAA receptors can be assembled from a combination of at least 16 different subunits. The receptor’s subunit composition determines its pharmacologic and functional properties, and subunit expression varies throughout the brain. A primary goal for new treatments targeting GABAA receptors is the production of subunit-selective modulators acting upon a discrete population of receptors. The anxiolytic 4-amino-7-hydroxy-2-methyl-5,6,7,8,-tetrahydrobenzo[b]thieno[2,3-b]pyridine-3-carboxylic acid, but-2-ynyl ester (SB-205384) is widely considered to be selective for α3-containing GABAA receptors. However, it has been tested only on α1-, α2-, and α3-containing receptors. We examined the activity of SB-205384 at recombinant receptors containing the six different α subunits and found that receptors containing the α3, α5, and α6 subunits were potentiated by SB-205384, with the α6 subunit conferring the greatest responsiveness. Properties associated with chimeric α1/α6 subunits suggested that multiple structural domains influence sensitivity to SB-205384. Point mutations of residues within the extracellular N-terminal domain identified a leucine residue located in loop E of the agonist binding site as an important determinant of high sensitivity to modulation. In the α6 subunit the identity of this residue is species-dependent, with the leucine found in rat subunits but not in human. Our results indicate that SB-205384 is not an α3-selective modulator, and instead acts at several GABAA receptor isoforms. These findings have implications for the side-effect profile of this anxiolytic as well as for its use in neuronal and animal studies as a marker for contribution from α3-containing receptors. PMID:23902941

DNA sequences, recombinant DNA molecules and processes for producing the A and B subunits of cholera toxin and preparations containing so-obtained subunit or subunits

DOE Office of Scientific and Technical Information (OSTI.GOV)

Harford, N.; De Wilde, M.

1987-05-19

A recombinant DNA molecule is described comprising at least a portion coding for subunits A and B of cholera toxin, or a fragment or derivative of the portion wherein the fragment or derivative codes for a polypeptide have an activity which can induce an immune response to subunit A; can induce an immune response to subunit A and cause epithelial cell penetration and the enzymatic effect leading to net loss of fluid into the gut lumen; can bind to the membrane receptor for the B subunit of cholera toxin; can induce an immune response to subunit B; can induce anmore » immune response to subunit B and bind to the membrane receptor; or has a combination of the activities.« less

Sucrose regulation of ADP-glucose pyrophosphorylase subunit genes transcript levels in leaves and fruits

NASA Technical Reports Server (NTRS)

Li, Xiangyang; Xing, Jinpeng; Gianfagna, Thomas J.; Janes, Harry W.

2002-01-01

ADP-glucose pyrophosphorylase (AGPase, EC2.7.7.27) is a key regulatory enzyme in starch biosynthesis. The enzyme is a heterotetramer with two S and two B subunits. In tomato, there are three multiple forms of the S subunit gene. Agp S1, S2 and B are highly expressed in fruit from 10 to 25 days after anthesis. Agp S3 is only weakly expressed in fruit. Sucrose significantly elevates expression of Agp S1, S2 and B in both leaves and fruits. Agp S1 exhibits the highest degree of regulation by sucrose. In fact, sucrose may be required for Agp S1 expression. For excised leaves incubated in water, no transcripts for Agp S1 could be detected in the absence of sucrose, whereas it took up to 16 h in water before transcripts were no longer detectable for Agp S2 and B. Neither Agp S3 nor the tubulin gene is affected by sucrose, demonstrating that this response is specifically regulated by a carbohydrate metabolic signal, and is not due to a general increase in metabolism caused by sucrose treatment. Truncated versions of the promoter for Agp S1 indicate that a specific region 1.3-3.0 kb upstream from the transcription site is responsible for sucrose sensitivity. This region of the S1 promoter contains several cis-acting elements present in the promoters of other genes that are also regulated by sucrose. c2002 Elsevier Science Ireland Ltd. All rights reserved.

2001 Wisconsin traffic crash facts

DOT National Transportation Integrated Search

2002-09-01

2001 Facts and Figures: 764 persons were killed in Wisconsin motor vehicle traffic crashes (40% involved alcohol, 32% involved : speed, and 16% involved both speed and alcohol). : 58,279 persons were injured in 39,358 reported injury crashes and ...

Cross-linking of initiation factor IF3 to Escherichia coli 30S ribosomal subunit by trans-diamminedichloroplatinum(II): characterization of two cross-linking sites in 16S rRNA; a possible way of functioning for IF3.

PubMed Central

Ehresmann, C; Moine, H; Mougel, M; Dondon, J; Grunberg-Manago, M; Ebel, J P; Ehresmann, B

1986-01-01

The initiation factor IF3 is platinated with trans-diamminedichloroplatinum(II) and cross-linked to Escherichia coli 30S ribosomal subunit. Two cross-linking sites are unambiguously identified on the 16S rRNA: a major one, in the region 819-859 in the central domain, and a minor one, in the region 1506-1529 in the 3'-terminal domain. Specific features of these sequences together with their particular location within the 30S subunit lead us to postulate a role for IF3, that conciliates topographical and functional observations made so far. Images PMID:2425339

Localization of nuclear subunits of cyclic AMP-dependent protein kinase by the immunocolloidal gold method

PubMed Central

1985-01-01

An immunocolloidal gold electron microscopy method is described allowing the ultrastructural localization and quantitation of the regulatory subunits RI and RII and the catalytic subunit C of cAMP- dependent protein kinase. Using a postembedding indirect immunogold labeling procedure that employs specific antisera, the catalytic and regulatory subunits were localized in electron-dense regions of the nucleus and in cytoplasmic areas with a minimum of nonspecific staining. Antigenic domains were localized in regions of the heterochromatin, nucleolus, interchromatin granules, and in the endoplasmic reticulum of different cell types, such as rat hepatocytes, ovarian granulosa cells, and spermatogonia, as well as cultured H4IIE hepatoma cells. Morphometric quantitation of the relative staining density of nuclear antigens indicated a marked modulation of the number of subunits per unit area under various physiologic conditions. For instance, following partial hepatectomy in rats, the staining density of the nuclear RI and C subunits was markedly increased 16 h after surgery. Glucagon treatment of rats increased the staining density of only the nuclear catalytic subunit. Dibutyryl cAMP treatment of H4IIE hepatoma cells led to a marked increase in the nuclear staining density of all three subunits of cAMP-dependent protein kinase. These studies demonstrate that specific antisera against cAMP-dependent protein kinase subunits may be used in combination with immunogold electron microscopy to identify the ultrastructural location of the subunits and to provide a semi-quantitative estimate of their relative cellular density. PMID:2993318

Monster Clusters in the Young Universe? Weak-lensing Masses of SPT-CL J0205-5829 and MOO1014+0038 with HST Observations

NASA Astrophysics Data System (ADS)

Feilx Kim, Seojin; Jee, Myungkook James

2018-01-01

Measuring High-z clusters’ masses is very important as the cluster abundance is extremely sensitive to the cosmological parameters. However, deriving their masses from the intracluster medium properties (i.e., Sunyaev-Zel’dovich or X-ray observations) is not the best method because of their departure from the hydrostatic equilibrium. Fortunately, the “See Change” Hubble Space Telescope program offers a rare opportunity to measure them using weak gravitational lensing. We study SPT-CL J0205-5829 (z=1.322) and MOO1014+0038 (z=1.24) discovered in the SPT-SZ and MaDCoW Surveys, respectively. Previous non-lensing based approaches suggest that both targets might be extremely massive clusters. After carefully addressing various possible systematics from the Advanced Camera for Surveys (ACS) and Wide Field Camera 3 (WFC3) images, we successfully detect clear weak lensing signals. We present their 2-dimensional mass maps and compare our weak-lensing masses with previous ICM-based results.

COX16 promotes COX2 metallation and assembly during respiratory complex IV biogenesis

PubMed Central

Aich, Abhishek; Wang, Cong; Chowdhury, Arpita; Ronsör, Christin; Pacheu-Grau, David; Richter-Dennerlein, Ricarda; Dennerlein, Sven

2018-01-01

Cytochrome c oxidase of the mitochondrial oxidative phosphorylation system reduces molecular oxygen with redox equivalent-derived electrons. The conserved mitochondrial-encoded COX1- and COX2-subunits are the heme- and copper-center containing core subunits that catalyze water formation. COX1 and COX2 initially follow independent biogenesis pathways creating assembly modules with subunit-specific, chaperone-like assembly factors that assist in redox centers formation. Here, we find that COX16, a protein required for cytochrome c oxidase assembly, interacts specifically with newly synthesized COX2 and its copper center-forming metallochaperones SCO1, SCO2, and COA6. The recruitment of SCO1 to the COX2-module is COX16- dependent and patient-mimicking mutations in SCO1 affect interaction with COX16. These findings implicate COX16 in CuA-site formation. Surprisingly, COX16 is also found in COX1-containing assembly intermediates and COX2 recruitment to COX1. We conclude that COX16 participates in merging the COX1 and COX2 assembly lines. PMID:29381136

GABAA receptors: Various stoichiometrics of subunit arrangement in α1β3 and α1β3ε receptors.

PubMed

Has, Ahmad Tarmizi Che; Chebib, Mary

2018-05-15

GABAA receptors (GABAARs) are members of the Cys-loop ligand-gated ion channel (LGIC) superfamily, which includes nicotinic acetylcholine, glycine, and serotonin (5HT3) receptors [1,2,3,4]. LGICs typically mediate fast synaptic transmission via the movement of ions through channels gated by neurotransmitters, such as acetylcholine for nicotinic receptors and GABA for GABAARs [5]. The term Cys-loop receptors originates from the presence of a conserved disulphide bond (or bridge) which holds together two cysteine amino acids of the loop that forms from the structure of polypeptides in the extracellular domain of the receptor's subunit [6]. GABAARs are pentameric transmembrane protein complexes consisting of five subunits from a variety of polypeptide subunits [7,8]. All of these subunits are pseudo-symmetrically organized in the plane of the membrane, with a Cl--selective channel in the middle of the complex. To date, nineteen GABAAR subunits have been identified and categorized into eight classes, α1-6, β1-3, γ1-3, δ, ε, θ, π and ρ1-3, but their variety is further broadened by the existence of several splice forms for certain subunits (e.g., α6, β2 and γ2) [9,10,11,12]. The subunits within each class have an amino acid sequence homology of 70% or more, whereas those with a sequence homology of 30% or less are grouped into different classes [13,14]. A subunit consists of four transmembrane domains (TM1-TM4), each forming an α-helix; a large extracellular N-terminal domain that incorporates part of the orthosteric agonist/antagonist binding site; a large intracellular loop between the TM3 and TM4; a small intracellular loop between TM1 and TM2; a small extracellular loop between TM2 and TM3; and a C-terminal extracellular domain [15,16]. Each subunit is arranged in such a way as to create principal and complementary interfaces with the other subunits, and in a position such that the TM2 of each subunit forms the wall of the channel pore [17,18,19]. The

Alpha subunit of glycoprotein hormones in the sera of acromegalic patients and its mRNA in the tumors.

PubMed

Machiavelli, G A; Artese, R; Benencia, H; Bruno, O; Guerra, L; Basso, A; Burdman, J A

1999-04-01

Within a population of 16 pituitary adenomas we found high levels of glycoprotein alpha subunits in the sera of patients with somatotrophic tumors. This finding was correlated with the presence of mRNA alpha subunit in these tumors indicating the adenomas themselves as the origin of the circulating alpha-subunit. Synthesis of these two hormones, which are chemically very different, by the same tumor cells indicates a high degree of differentiation of these cells. We are unable at this time to conclusively correlate differentiation of these tumors aggressively.

Spectroscopic Confirmation of Five Galaxy Clusters at z > 1.25 in the 2500 deg^2 SPT-SZ Survey

NASA Astrophysics Data System (ADS)

Khullar, Gourav; Bleem, Lindsey; Bayliss, Matthew; Gladders, Michael; South Pole Telescope (SPT) Collaboration

2018-06-01

We present spectroscopic confirmation of 5 galaxy clusters at 1.25 < z < 1.5, discovered in the 2500 deg2 South Pole Telescope Sunyaev-Zel’dovich (SPT-SZ) survey. These clusters, taken from a nearly redshift-independent mass-limited sample of clusters, have multi-wavelength follow-up imaging data from the X-ray to the near-IR, and currently form the most homogenous massive high-redshift cluster sample in existence. We briefly describe the analysis pipeline used on the low S/N spectra of these faint galaxies, and describing the multiple techniques used to extract robust redshifts from a combination of absorption-line (Ca II H&K doublet - λλ3934,3968Å) and emission-line ([OII] λλ3727,3729Å) spectral features. We present several ensemble analyses of cluster member galaxies that demonstrate the reliability of the measured redshifts. We also identify modest [OII] emission and pronounced CN and Hδ absorption in a composite stacked spectrum of 28 low S/N passive galaxy spectra with redshifts derived primarily from Ca II H&K features. This work increases the number of spectroscopically-confirmed SPT-SZ galaxy clusters at z > 1.25 from 2 to 7, further demonstrating the efficacy of SZ selection for the highest redshift massive clusters, and enabling further detailed study of these confirmed systems.

Constraints on Cosmological Parameters from the Angular Power Spectrum of a Combined 2500 deg2 SPT-SZ and Planck Gravitational Lensing Map

NASA Astrophysics Data System (ADS)

Simard, G.; Omori, Y.; Aylor, K.; Baxter, E. J.; Benson, B. A.; Bleem, L. E.; Carlstrom, J. E.; Chang, C. L.; Cho, H.-M.; Chown, R.; Crawford, T. M.; Crites, A. T.; de Haan, T.; Dobbs, M. A.; Everett, W. B.; George, E. M.; Halverson, N. W.; Harrington, N. L.; Henning, J. W.; Holder, G. P.; Hou, Z.; Holzapfel, W. L.; Hrubes, J. D.; Knox, L.; Lee, A. T.; Leitch, E. M.; Luong-Van, D.; Manzotti, A.; McMahon, J. J.; Meyer, S. S.; Mocanu, L. M.; Mohr, J. J.; Natoli, T.; Padin, S.; Pryke, C.; Reichardt, C. L.; Ruhl, J. E.; Sayre, J. T.; Schaffer, K. K.; Shirokoff, E.; Staniszewski, Z.; Stark, A. A.; Story, K. T.; Vanderlinde, K.; Vieira, J. D.; Williamson, R.; Wu, W. L. K.

2018-06-01

We report constraints on cosmological parameters from the angular power spectrum of a cosmic microwave background (CMB) gravitational lensing potential map created using temperature data from 2500 deg2 of South Pole Telescope (SPT) data supplemented with data from Planck in the same sky region, with the statistical power in the combined map primarily from the SPT data. We fit the lensing power spectrum to a model including cold dark matter and a cosmological constant ({{Λ }}{CDM}), and to models with single-parameter extensions to {{Λ }}{CDM}. We find constraints that are comparable to and consistent with those found using the full-sky Planck CMB lensing data, e.g., {σ }8{{{Ω }}}{{m}}0.25 = 0.598 ± 0.024 from the lensing data alone with weak priors placed on other parameters. Combining with primary CMB data, we explore single-parameter extensions to {{Λ }}{CDM}. We find {{{Ω }}}k =-{0.012}-0.023+0.021 or {M}ν < 0.70 eV at 95% confidence, in good agreement with results including the lensing potential as measured by Planck. We include two parameters that scale the effect of lensing on the CMB: {A}L, which scales the lensing power spectrum in both the lens reconstruction power and in the smearing of the acoustic peaks, and {A}φ φ , which scales only the amplitude of the lensing reconstruction power spectrum. We find {A}φ φ × {A}L = 1.01 ± 0.08 for the lensing map made from combined SPT and Planck data, indicating that the amount of lensing is in excellent agreement with expectations from the observed CMB angular power spectrum when not including the information from smearing of the acoustic peaks.

Constraints on Cosmological Parameters from the Angular Power Spectrum of a Combined 2500 deg$^2$ SPT-SZ and Planck Gravitational Lensing Map

DOE PAGES

Simard, G.; et al.

2018-06-20

We report constraints on cosmological parameters from the angular power spectrum of a cosmic microwave background (CMB) gravitational lensing potential map created using temperature data from 2500 degmore » $^2$ of South Pole Telescope (SPT) data supplemented with data from Planck in the same sky region, with the statistical power in the combined map primarily from the SPT data. We fit the corresponding lensing angular power spectrum to a model including cold dark matter and a cosmological constant ($$\\Lambda$$CDM), and to models with single-parameter extensions to $$\\Lambda$$CDM. We find constraints that are comparable to and consistent with constraints found using the full-sky Planck CMB lensing data. Specifically, we find $$\\sigma_8 \\Omega_{\\rm m}^{0.25}=0.598 \\pm 0.024$$ from the lensing data alone with relatively weak priors placed on the other $$\\Lambda$$CDM parameters. In combination with primary CMB data from Planck, we explore single-parameter extensions to the $$\\Lambda$$CDM model. We find $$\\Omega_k = -0.012^{+0.021}_{-0.023}$$ or $$M_{\

Constraints on Cosmological Parameters from the Angular Power Spectrum of a Combined 2500 deg$^2$ SPT-SZ and Planck Gravitational Lensing Map

DOE Office of Scientific and Technical Information (OSTI.GOV)

Simard, G.; et al.

We report constraints on cosmological parameters from the angular power spectrum of a cosmic microwave background (CMB) gravitational lensing potential map created using temperature data from 2500 degmore » $^2$ of South Pole Telescope (SPT) data supplemented with data from Planck in the same sky region, with the statistical power in the combined map primarily from the SPT data. We fit the corresponding lensing angular power spectrum to a model including cold dark matter and a cosmological constant ($$\\Lambda$$CDM), and to models with single-parameter extensions to $$\\Lambda$$CDM. We find constraints that are comparable to and consistent with constraints found using the full-sky Planck CMB lensing data. Specifically, we find $$\\sigma_8 \\Omega_{\\rm m}^{0.25}=0.598 \\pm 0.024$$ from the lensing data alone with relatively weak priors placed on the other $$\\Lambda$$CDM parameters. In combination with primary CMB data from Planck, we explore single-parameter extensions to the $$\\Lambda$$CDM model. We find $$\\Omega_k = -0.012^{+0.021}_{-0.023}$$ or $$M_{\

Identification of a small TAF complex and its role in the assembly of TAF-containing complexes.

PubMed

Demény, Màté A; Soutoglou, Evi; Nagy, Zita; Scheer, Elisabeth; Jànoshàzi, Agnes; Richardot, Magalie; Argentini, Manuela; Kessler, Pascal; Tora, Laszlo

2007-03-21

TFIID plays a role in nucleating RNA polymerase II preinitiation complex assembly on protein-coding genes. TFIID is a multisubunit complex comprised of the TATA box binding protein (TBP) and 14 TBP-associated factors (TAFs). Another class of multiprotein transcriptional regulatory complexes having histone acetyl transferase (HAT) activity, and containing TAFs, includes TFTC, STAGA and the PCAF/GCN5 complex. Looking for as yet undiscovered subunits by a proteomic approach, we had identified TAF8 and SPT7L in human TFTC preparations. Subsequently, however, we demonstrated that TAF8 was not a stable component of TFTC, but that it is present in a small TAF complex (SMAT), containing TAF8, TAF10 and SPT7L, that co-purified with TFTC. Thus, TAF8 is a subunit of both TFIID and SMAT. The latter has to be involved in a pathway of complex formation distinct from the other known TAF complexes, since these three histone fold (HF)-containing proteins (TAF8, TAF10 and SPT7L) can never be found together either in TFIID or in STAGA/TFTC HAT complexes. Here we show that TAF8 is absolutely necessary for the integration of TAF10 in a higher order TFIID core complex containing seven TAFs. TAF8 forms a heterodimer with TAF10 through its HF and proline rich domains, and also interacts with SPT7L through its C-terminal region, and the three proteins form a complex in vitro and in vivo. Thus, the TAF8-TAF10 and TAF10-SPT7L HF pairs, and also the SMAT complex, seem to be important regulators of the composition of different TFIID and/or STAGA/TFTC complexes in the nucleus and consequently may play a role in gene regulation.

20 Facts on Women Workers. Facts on Working Women No. 90-2.

ERIC Educational Resources Information Center

Women's Bureau (DOL), Washington, DC.

This information sheet presents 20 facts on women workers in 1989: (1) 56 million women 16 years of age and over are working or looking for work; (2) 69 percent of all women 18 to 64 years of age are in the civilian labor force; (3) most women workers are employed full time; (4) the average women worker spends 29.3 years of her life in the labor…

The sodium pump alpha1 subunit as a potential target to combat apoptosis-resistant glioblastomas.

PubMed

Lefranc, Florence; Kiss, Robert

2008-03-01

To review the involvement of the ion transporter Na+/K+-ATPase (NaK) in the migration and proliferation of glioma cells. Preliminary studies indicate that NaK alpha1 subunits seem to be upregulated in a proportion of glioblastomas but not in normal brain tissues. The present review focuses on (1) the natural resistance of migrating malignant glioma cells to apoptosis, (2) autophagic cell death as an alternative to combat malignant gliomas, (3) the fact that reducing the levels of malignant glioma cell motility can restore proapoptotic drug sensitivity,and (4) on the observation that inhibiting the NaK activity reduces both glioma cell proliferation and migration. The natural ligands of the NaK are the cardiotonic steroids. A hemisynthetic derivative of 2"-oxovoruscharin (UNBS1450), a novel cardenolide, displays unique structural features, making its binding affinity to NaK alpha subunits (including alpha1) 10 to 100 times higher than that of other cardenolides. UNBS1450 markedly decreases intracellular ATP concentration in glioma cells, disorganizes the actin cytoskeleton, and leads to autophagic cell death in NaK alpha1 over-expressing glioma cells. Glioblastoma patients who do not respond to chemotherapy and whose tumors over-express NaK alpha1 subunits could benefit from a treatment using ligands with marked binding affinity for the NaK alpha1 subunit.

Ssrp1a controls organogenesis by promoting cell cycle progression and RNA synthesis.

PubMed

Koltowska, Katarzyna; Apitz, Holger; Stamataki, Despina; Hirst, Elizabeth M A; Verkade, Heather; Salecker, Iris; Ober, Elke A

2013-05-01

Tightly controlled DNA replication and RNA transcription are essential for differentiation and tissue growth in multicellular organisms. Histone chaperones, including the FACT (facilitates chromatin transcription) complex, are central for these processes and act by mediating DNA access through nucleosome reorganisation. However, their roles in vertebrate organogenesis are poorly understood. Here, we report the identification of zebrafish mutants for the gene encoding Structure specific recognition protein 1a (Ssrp1a), which, together with Spt16, forms the FACT heterodimer. Focussing on the liver and eye, we show that zygotic Ssrp1a is essential for proliferation and differentiation during organogenesis. Specifically, gene expression indicative of progressive organ differentiation is disrupted and RNA transcription is globally reduced. Ssrp1a-deficient embryos exhibit DNA synthesis defects and prolonged S phase, uncovering a role distinct from that of Spt16, which promotes G1 phase progression. Gene deletion/replacement experiments in Drosophila show that Ssrp1b, Ssrp1a and N-terminal Ssrp1a, equivalent to the yeast homologue Pob3, can substitute Drosophila Ssrp function. These data suggest that (1) Ssrp1b does not compensate for Ssrp1a loss in the zebrafish embryo, probably owing to insufficient expression levels, and (2) despite fundamental structural differences, the mechanisms mediating DNA accessibility by FACT are conserved between yeast and metazoans. We propose that the essential functions of Ssrp1a in DNA replication and gene transcription, together with its dynamic spatiotemporal expression, ensure organ-specific differentiation and proportional growth, which are crucial for the forming embryo.

Identification of dietary alanine toxicity and trafficking dysfunction in a Drosophila model of hereditary sensory and autonomic neuropathy type 1

PubMed Central

Oswald, Matthew C. W.; West, Ryan J. H.; Lloyd-Evans, Emyr; Sweeney, Sean T.

2015-01-01

Hereditary sensory and autonomic neuropathy type 1 (HSAN1) is characterized by a loss of distal peripheral sensory and motorneuronal function, neuropathic pain and tissue necrosis. The most common cause of HSAN1 is due to dominant mutations in serine palmitoyl-transferase subunit 1 (SPT1). SPT catalyses the condensation of serine with palmitoyl-CoA, the initial step in sphingolipid biogenesis. Identified mutations in SPT1 are known to both reduce sphingolipid synthesis and generate catalytic promiscuity, incorporating alanine or glycine into the precursor sphingolipid to generate a deoxysphingoid base (DSB). Why either loss of function in SPT1, or generation of DSBs should generate deficits in distal sensory function remains unclear. To address these questions, we generated a Drosophila model of HSAN1. Expression of dSpt1 bearing a disease-related mutation induced morphological deficits in synapse growth at the larval neuromuscular junction consistent with a dominant-negative action. Expression of mutant dSpt1 globally was found to be mildly toxic, but was completely toxic when the diet was supplemented with alanine, when DSBs were observed in abundance. Expression of mutant dSpt1 in sensory neurons generated developmental deficits in dendritic arborization with concomitant sensory deficits. A membrane trafficking defect was observed in soma of sensory neurons expressing mutant dSpt1, consistent with endoplasmic reticulum (ER) to Golgi block. We found that we could rescue sensory function in neurons expressing mutant dSpt1 by co-expressing an effector of ER–Golgi function, Rab1 suggesting compromised ER function in HSAN1 affected dendritic neurons. Our Drosophila model identifies a novel strategy to explore the pathological mechanisms of HSAN1. PMID:26395456

Assembly of the stator in Escherichia coli ATP synthase. Complexation of alpha subunit with other F1 subunits is prerequisite for delta subunit binding to the N-terminal region of alpha

PubMed Central

Senior, Alan E.; Muharemagi, Alma; Wilke-Mounts, Susan

2008-01-01

Alpha subunit of Escherichia coli ATP synthase was expressed with a C-terminal 6-His tag and purified. Pure alpha was monomeric, competent in nucleotide binding, and had normal N-terminal sequence. In F1-subunit dissociation/reassociation experiments it supported full reconstitution of ATPase, and reassociated complexes were able to bind to F1-depleted membranes with restoration of ATP-driven proton pumping. Therefore interaction between the stator delta subunit and the N-terminal residue 1-22 region of alpha occurred normally when pure alpha was complexed with other F1 subunits. On the other hand, three different types of experiment showed that no interaction occurred between pure delta and isolated alpha subunit. Unlike in F1, the N-terminal region of isolated alpha was not susceptible to trypsin cleavage. Therefore, during assembly of ATP synthase, complexation of alpha subunit with other F1 subunits is prerequisite for delta subunit binding to the N-terminal region of alpha. We suggest that the N-terminal 1-22 residues of alpha are sequestered in isolated alpha until released by binding of beta to alpha subunit. This prevents 1/1 delta/alpha complexes from forming, and provides a satisfactory explanation of the stoichiometry of one delta per three alpha seen in the F1 sector of ATP synthase, assuming that steric hindrance prevents binding of more than one delta to the alpha3/beta3 hexagon. The cytoplasmic fragment of the b subunit (bsol) did not bind to isolated alpha. It might also be that complexation of alpha with beta subunits is prerequisite for direct binding of stator b subunit to the F1-sector. PMID:17176112

Evaluating facts and facting evaluations: On the fact-value relationship in HTA.

PubMed

Hofmann, Bjørn; Bond, Ken; Sandman, Lars

2018-04-03

Health technology assessment (HTA) is an evaluation of health technologies in terms of facts and evidence. However, the relationship between facts and values is still not clear in HTA. This is problematic in an era of "fake facts" and "truth production." Accordingly, the objective of this study is to clarify the relationship between facts and values in HTA. We start with the perspectives of the traditional positivist account of "evaluating facts" and the social-constructivist account of "facting values." Our analysis reveals diverse relationships between facts and a spectrum of values, ranging from basic human values, to the values of health professionals, and values of and in HTA, as well as for decision making. We argue for sensitivity to the relationship between facts and values on all levels of HTA, for being open and transparent about the values guiding the production of facts, and for a primacy for the values close to the principal goals of health care, ie, relieving suffering. We maintain that philosophy (in particular ethics) may have an important role in addressing the relationship between facts and values in HTA. Philosophy may help us to avoid fallacies of inferring values from facts; to disentangle the normative assumptions in the production or presentation of facts and to tease out implicit value judgements in HTA; to analyse evaluative argumentation relating to facts about technologies; to address conceptual issues of normative importance; and to promote reflection on HTA's own value system. In this we argue for a(n Aristotelian) middle way between the traditional positivist account of "evaluating facts" and the social-constructivist account of "facting values," which we call "factuation." We conclude that HTA is unique in bringing together facts and values and that being conscious and explicit about this "factuation" is key to making HTA valuable to both individual decision makers and society as a whole. © 2018 The Authors Journal of Evaluation in

Stellar Masses and Star Formation Rates of Lensed, Dusty, Star-forming Galaxies from the SPT Survey

NASA Astrophysics Data System (ADS)

Ma, Jingzhe; Gonzalez, Anthony. H.; Spilker, J. S.; Strandet, M.; Ashby, M. L. N.; Aravena, M.; Béthermin, M.; Bothwell, M. S.; de Breuck, C.; Brodwin, M.; Chapman, S. C.; Fassnacht, C. D.; Greve, T. R.; Gullberg, B.; Hezaveh, Y.; Malkan, M.; Marrone, D. P.; Saliwanchik, B. R.; Vieira, J. D.; Weiss, A.; Welikala, N.

2015-10-01

To understand cosmic mass assembly in the universe at early epochs, we primarily rely on measurements of the stellar masses and star formation rates (SFRs) of distant galaxies. In this paper, we present stellar masses and SFRs of six high-redshift (2.8 ≤ z ≤ 5.7) dusty, star-forming galaxies (DSFGs) that are strongly gravitationally lensed by foreground galaxies. These sources were first discovered by the South Pole Telescope (SPT) at millimeter wavelengths and all have spectroscopic redshifts and robust lens models derived from Atacama Large Millimeter/submillimeter Array observations. We have conducted follow-up observations to obtain multi-wavelength imaging data using the Hubble Space Telescope (HST), Spitzer, Herschel, and the Atacama Pathfinder EXperiment. We use the high-resolution HST/Wide Field Camera 3 images to disentangle the background source from the foreground lens in Spitzer/IRAC data. The detections and upper limits provide important constraints on the spectral energy distributions (SEDs) for these DSFGs, yielding stellar masses, IR luminosities, and SFRs. The SED fits of six SPT sources show that the intrinsic stellar masses span a range more than one order of magnitude with a median value ˜5 ×1010 M⊙. The intrinsic IR luminosities range from 4 × 1012 L⊙ to 4 × 1013 L⊙. They all have prodigious intrinsic SFRs of 510-4800 M⊙ yr-1. Compared to the star-forming main sequence (MS), these six DSFGs have specific SFRs that all lie above the MS, including two galaxies that are a factor of 10 higher than the MS. Our results suggest that we are witnessing ongoing strong starburst events that may be driven by major mergers.

Bacillus caseinilyticus sp. nov., an alkali- and thermotolerant bacterium isolated from a soda lake.

PubMed

Vishnuvardhan Reddy, Sultanpuram; Thirumala, Mothe; Farooq, Mohammed

2015-08-01

A novel Gram-stain-positive, rod-shaped, motile, endospore-forming and proteolytic bacterial strain, SPT, was isolated from Lonar soda lake, in India. On the basis of 16S rRNA gene sequence analysis it was identified as belonging to the class Firmibacteria and was most closely related to Bacillus cellulosilyticus DSM 2522T (96.7%) and other members of the genus Bacillus ( < 95.9%). Strain SPT was catalase- and oxidase-positive. The cell-wall peptidoglycan of strain SPT contained meso-diaminopimelic acid. Polar lipids included diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, three phospholipids, two aminolipids and two unknown lipids. The predominant isoprenoid quinone was MK-7. Anteiso-C15 : 0 (26.8%) was the predominant fatty acid and significant proportions (>5%) of iso-C15 : 0 (20.9%), C16 : 1ω7c alcohol (6.3%), iso-C16 : 0 (6.3%) and anteiso-C17 : 0 (5.3  %) were also detected in strain SPT. The DNA G+C content of strain SPT was 38.9 mol%. The results of phylogenetic, chemotaxonomic and biochemical tests allowed a clear differentiation of strain SPT from all other members of the genus Bacillus. Strain SPT represents a novel member of the genus Bacillus, for which the name Bacilluscaseinilyticus sp. nov. is proposed. The type strain is SPT ( = MCC 2612T = JCM 30246T).

The Sodium Pump α1 Subunit as a Potential Target to Combat Apoptosis-Resistant Glioblastomas1

PubMed Central

Lefranc, Florence; Kiss, Robert

2008-01-01

Purpose To review the involvement of the ion transporter Na+/K+-ATPase (NaK) in the migration and proliferation of glioma cells. Preliminary studies indicate that NaK α1 subunits seem to be upregulated in a proportion of glioblastomas but not in normal brain tissues. Design The present review focuses on (1) the natural resistance of migrating malignant glioma cells to apoptosis, (2) autophagic cell death as an alternative to combat malignant gliomas, (3) the fact that reducing the levels of malignant glioma cell motility can restore proapoptotic drug sensitivity, and (4) on the observation that inhibiting the NaK activity reduces both glioma cell proliferation and migration. Results The natural ligands of the NaK are the cardiotonic steroids. A hemisynthetic derivative of 2″-oxovoruscharin (UNBS1450), a novel cardenolide, displays unique structural features, making its binding affinity to NaK α subunits (including α1) 10 to 100 times higher than that of other cardenolides. UNBS1450 markedly decreases intracellular ATP concentration in glioma cells, disorganizes the actin cytoskeleton, and leads to autophagic cell death in NaK α1 over-expressing glioma cells. Conclusions Glioblastoma patients who do not respond to chemotherapy and whose tumors over-express NaK α1 subunits could benefit from a treatment using ligands with marked binding affinity for the NaK α1 subunit. PMID:18323016

Na+/K+-ATPase α-subunit in swimming crab Portunus trituberculatus: molecular cloning, characterization, and expression under low salinity stress

NASA Astrophysics Data System (ADS)

Han, Xiaolin; Liu, Ping; Gao, Baoquan; Wang, Haofeng; Duan, Yafei; Xu, Wenfei; Chen, Ping

2015-07-01

Na+/K+-ATPases are membrane-associated enzymes responsible for the active transport of Na+ and K+ ions across cell membranes, generating chemical and electrical gradients. These enzymes' α-subunit provides catalytic function, binding and hydrolyzing ATP, and itself becoming phosphorylated during the transport cycle. In this study, Na+/K+-ATPase α-subunit cDNA was cloned from gill tissue of the swimming crab Portunus trituberculatus by reverse-transcription polymerase chain reaction (RT-PCR) and rapid amplification of cDNA end methods. Analysis of the nucleotide sequence revealed that the cDNA had a full-length of 3 833 base pairs (bp), with an open reading frame of 3 120 bp, 5' untranslated region (UTR) of 317 bp, and 3' UTR of 396 bp. The sequence encoded a 1 039 amino acid protein with a predicted molecular weight of 115.57 kDa and with estimated pI of 5.21. It was predicted here to possess all expected features of Na+/K+-ATPase members, including eight transmembrane domains, putative ATP-binding site, and phosphorylation site. Comparison of amino acid sequences showed that the P. trituberculatus α-subunit possessed an overall identity of 75%-99% to that of other organisms. Phylogenetic analysis revealed that this α-subunit was in the same category as those of crustaceans. Quantitative real-time RT-PCR analysis indicated that this α-subunit's transcript were most highly expressed in gill and lowest in muscle. RT-PCR analysis also revealed that α-subunit expression in crab gill decreased after 2 and 6 h, but increased after 12, 24, 48, and 72 h. In addition, α-subunit expression in hepatopancreas of crab decreased after 2-72 h. These facts indicated that the crab's Na+/K+-ATPase α-subunit was potentially involved in the observed acute response to low salinity stress.

The role of a conserved membrane proximal cysteine in altering αPS2CβPS integrin diffusion.

PubMed

Syed, Aleem; Arora, Neha; Bunch, Thomas A; Smith, Emily A

2016-11-15

Cysteine residues (Cys) in the membrane proximal region are common post-translational modification (PTM) sites in transmembrane proteins. Herein, the effects of a highly conserved membrane proximal α-subunit Cys 1368 on the diffusion properties of αPS2CβPS integrins are reported. Sequence alignment shows that this cysteine is palmitoylated in human α3 and α6 integrin subunits. Replacing Cys 1368 in wild-type integrins with valine (Val 1368 ) putatively blocks a PTM site and alters integrins' ligand binding and diffusion characteristics. Both fluorescence recovery after photobleaching (FRAP) and single particle tracking (SPT) diffusion measurements show Val 1368 integrins are more mobile compared to wild-type integrins. Approximately 33% and 8% more Val 1368 integrins are mobile as measured by FRAP and SPT, respectively. The mobile Val 1368 integrins also exhibit less time-dependent diffusion, as measured by FRAP. Tandem mass spectrometry data suggest that Cys 1368 contains a redox or palmitoylation PTM in αPS2CβPS integrins. This membrane proximal Cys may play an important role in the diffusion of other alpha subunits that contain this conserved residue.

The role of a conserved membrane proximal cysteine in altering αPS2CβPS integrin diffusion

NASA Astrophysics Data System (ADS)

Syed, Aleem; Arora, Neha; Bunch, Thomas A.; Smith, Emily A.

2016-12-01

Cysteine residues (Cys) in the membrane proximal region are common post-translational modification (PTM) sites in transmembrane proteins. Herein, the effects of a highly conserved membrane proximal α-subunit Cys1368 on the diffusion properties of αPS2CβPS integrins are reported. Sequence alignment shows that this cysteine is palmitoylated in human α3 and α6 integrin subunits. Replacing Cys1368 in wild-type integrins with valine (Val1368) putatively blocks a PTM site and alters integrins’ ligand binding and diffusion characteristics. Both fluorescence recovery after photobleaching (FRAP) and single particle tracking (SPT) diffusion measurements show Val1368 integrins are more mobile compared to wild-type integrins. Approximately 33% and 8% more Val1368 integrins are mobile as measured by FRAP and SPT, respectively. The mobile Val1368 integrins also exhibit less time-dependent diffusion, as measured by FRAP. Tandem mass spectrometry data suggest that Cys1368 contains a redox or palmitoylation PTM in αPS2CβPS integrins. This membrane proximal Cys may play an important role in the diffusion of other alpha subunits that contain this conserved residue.

Expression of specific ionotropic glutamate and GABA-A receptor subunits is decreased in central amygdala of alcoholics.

PubMed

Jin, Zhe; Bhandage, Amol K; Bazov, Igor; Kononenko, Olga; Bakalkin, Georgy; Korpi, Esa R; Birnir, Bryndis

2014-01-01

The central amygdala (CeA) has a role for mediating fear and anxiety responses. It is also involved in emotional imbalance caused by alcohol abuse and dependence and in regulating relapse to alcohol abuse. Growing evidences suggest that excitatory glutamatergic and inhibitory γ-aminobutyric acid-ergic (GABAergic) transmissions in the CeA are affected by chronic alcohol exposure. Human post-mortem CeA samples from male alcoholics (n = 9) and matched controls (n = 9) were assayed for the expression level of ionotropic glutamate and GABA-A receptors subunit mRNAs using quantitative real-time reverse transcription-PCR (RT-qPCR). Our data revealed that out of the 16 ionotropic glutamate receptor subunits, mRNAs encoding two AMPA [2-amino-3-(3-hydroxy-5-methyl-isoxazol-4-yl)propanoic acid] receptor subunits GluA1 and GluA4; one kainate receptor subunit GluK2; one NMDA (N-methyl-D-aspartate) receptor subunit GluN2D and one delta receptor subunit GluD2 were significantly decreased in the CeA of alcoholics. In contrast, of the 19 GABA-A receptor subunits, only the mRNA encoding the α2 subunit was significantly down-regulated in the CeA of the alcoholics as compared with control subjects. Our findings imply that the down-regulation of specific ionotropic glutamate and GABA-A receptor subunits in the CeA of alcoholics may represent one of the molecular substrates underlying the new balance between excitatory and inhibitory neurotransmission in alcohol dependence.

Expression of specific ionotropic glutamate and GABA-A receptor subunits is decreased in central amygdala of alcoholics

PubMed Central

Jin, Zhe; Bhandage, Amol K.; Bazov, Igor; Kononenko, Olga; Bakalkin, Georgy; Korpi, Esa R.; Birnir, Bryndis

2014-01-01

The central amygdala (CeA) has a role for mediating fear and anxiety responses. It is also involved in emotional imbalance caused by alcohol abuse and dependence and in regulating relapse to alcohol abuse. Growing evidences suggest that excitatory glutamatergic and inhibitory γ-aminobutyric acid-ergic (GABAergic) transmissions in the CeA are affected by chronic alcohol exposure. Human post-mortem CeA samples from male alcoholics (n = 9) and matched controls (n = 9) were assayed for the expression level of ionotropic glutamate and GABA-A receptors subunit mRNAs using quantitative real-time reverse transcription-PCR (RT-qPCR). Our data revealed that out of the 16 ionotropic glutamate receptor subunits, mRNAs encoding two AMPA [2-amino-3-(3-hydroxy-5-methyl-isoxazol-4-yl)propanoic acid] receptor subunits GluA1 and GluA4; one kainate receptor subunit GluK2; one NMDA (N-methyl-D-aspartate) receptor subunit GluN2D and one delta receptor subunit GluD2 were significantly decreased in the CeA of alcoholics. In contrast, of the 19 GABA-A receptor subunits, only the mRNA encoding the α2 subunit was significantly down-regulated in the CeA of the alcoholics as compared with control subjects. Our findings imply that the down-regulation of specific ionotropic glutamate and GABA-A receptor subunits in the CeA of alcoholics may represent one of the molecular substrates underlying the new balance between excitatory and inhibitory neurotransmission in alcohol dependence. PMID:25278838

The SAGA/TREX-2 subunit Sus1 binds widely to transcribed genes and affects mRNA turnover globally.

PubMed

García-Molinero, Varinia; García-Martínez, José; Reja, Rohit; Furió-Tarí, Pedro; Antúnez, Oreto; Vinayachandran, Vinesh; Conesa, Ana; Pugh, B Franklin; Pérez-Ortín, José E; Rodríguez-Navarro, Susana

2018-03-29

Eukaryotic transcription is regulated through two complexes, the general transcription factor IID (TFIID) and the coactivator Spt-Ada-Gcn5 acetyltransferase (SAGA). Recent findings confirm that both TFIID and SAGA contribute to the synthesis of nearly all transcripts and are recruited genome-wide in yeast. However, how this broad recruitment confers selectivity under specific conditions remains an open question. Here we find that the SAGA/TREX-2 subunit Sus1 associates with upstream regulatory regions of many yeast genes and that heat shock drastically changes Sus1 binding. While Sus1 binding to TFIID-dominated genes is not affected by temperature, its recruitment to SAGA-dominated genes and RP genes is significantly disturbed under heat shock, with Sus1 relocated to environmental stress-responsive genes in these conditions. Moreover, in contrast to recent results showing that SAGA deubiquitinating enzyme Ubp8 is dispensable for RNA synthesis, genomic run-on experiments demonstrate that Sus1 contributes to synthesis and stability of a wide range of transcripts. Our study provides support for a model in which SAGA/TREX-2 factor Sus1 acts as a global transcriptional regulator in yeast but has differential activity at yeast genes as a function of their transcription rate or during stress conditions.

Subunit mass fingerprinting of mitochondrial complex I.

PubMed

Morgner, Nina; Zickermann, Volker; Kerscher, Stefan; Wittig, Ilka; Abdrakhmanova, Albina; Barth, Hans-Dieter; Brutschy, Bernhard; Brandt, Ulrich

2008-10-01

We have employed laser induced liquid bead ion desorption (LILBID) mass spectrometry to determine the total mass and to study the subunit composition of respiratory chain complex I from Yarrowia lipolytica. Using 5-10 pmol of purified complex I, we could assign all 40 known subunits of this membrane bound multiprotein complex to peaks in LILBID subunit fingerprint spectra by comparing predicted protein masses to observed ion masses. Notably, even the highly hydrophobic subunits encoded by the mitochondrial genome were easily detectable. Moreover, the LILBID approach allowed us to spot and correct several errors in the genome-derived protein sequences of complex I subunits. Typically, the masses of the individual subunits as determined by LILBID mass spectrometry were within 100 Da of the predicted values. For the first time, we demonstrate that LILBID spectrometry can be successfully applied to a complex I band eluted from a blue-native polyacrylamide gel, making small amounts of large multiprotein complexes accessible for subunit mass fingerprint analysis even if they are membrane bound. Thus, the LILBID subunit mass fingerprint method will be of great value for efficient proteomic analysis of complex I and its assembly intermediates, as well as of other water soluble and membrane bound multiprotein complexes.

Subunit arrangement in P2X receptors.

PubMed

Jiang, Lin-Hua; Kim, Miran; Spelta, Valeria; Bo, Xuenong; Surprenant, Annmarie; North, R Alan

2003-10-01

ATP-gated ionotropic receptors (P2X receptors) are distributed widely in the nervous system. For example, a hetero-oligomeric receptor containing both P2X2 and P2X3 subunits is involved in primary afferent sensation. Each subunit has two membrane-spanning domains. We have used disulfide bond formation between engineered cysteines to demonstrate close proximity between the outer ends of the first transmembrane domain of one subunit and the second transmembrane domain of another. After expression in HEK 293 cells of such modified P2X2 or P2X4 subunits, the disulfide bond formation is evident because an ATP-evoked channel opening requires previous reduction with dithiothreitol. In the hetero-oligomeric P2X2/3 receptor the coexpression of doubly substituted subunits with wild-type partners allows us to deduce that the hetero-oligomeric channel contains adjacent P2X3 subunits but does not contain adjacent P2X2 subunits. The results suggest a "head-to-tail" subunit arrangement in the quaternary structure of P2X receptors and show that a trimeric P2X2/3 receptor would have the composition P2X2(P2X3)2.

An analysis of subunit exchange in the dimeric DNA-binding and DNA-bending protein, TF1.

PubMed

Andera, L; Schneider, G J; Geiduschek, E P

1994-01-01

TF1 is the Bacillus subtilis bacteriophage-encoded dimeric type II DNA-binding protein. This relative of the eubacterial HU proteins and of the Escherichia coli integration host factor binds preferentially to 5-(hydroxymethyluracil)-containing DNA. We have examined the dynamics of exchange of monomer subunits between molecules of dimeric TF1. The analysis takes advantage of the fact that replacement of phenylalanine with arginine at amino acid 61 in the beta-loop 'arm' of TF1 alters DNA-bending and -binding properties, generating DNA complexes with distinctively different mobilities in gel electrophoresis. New species of DNA-protein complexes were formed by mixtures of wild type and mutant TF1, reflecting the formation of heterodimeric TF1, and making the dynamics of monomer exchange between TF1 dimers accessible to a simple gel retardation analysis. Exchange was rapid at high protein concentrations, even at 0 degrees C, and is proposed to be capable of proceeding through an interaction of molecules of TF1 dimer rather than exclusively through dissociation into monomer subunits. Evidence suggesting that DNA-bound TF1 dimers do not exchange subunits readily is also presented.

Mutant POLG2 Disrupts DNA Polymerase γ Subunits and Causes Progressive External Ophthalmoplegia

PubMed Central

Longley, Matthew J.; Clark, Susanna; Yu Wai Man, Cynthia; Hudson, Gavin; Durham, Steve E.; Taylor, Robert W.; Nightingale, Simon; Turnbull, Douglass M.; Copeland, William C.; Chinnery, Patrick F.

2006-01-01

DNA polymerase γ (pol γ) is required to maintain the genetic integrity of the 16,569-bp human mitochondrial genome (mtDNA). Mutation of the nuclear gene for the catalytic subunit of pol γ (POLG) has been linked to a wide range of mitochondrial diseases involving mutation, deletion, and depletion of mtDNA. We describe a heterozygous dominant mutation (c.1352G→A/p.G451E) in POLG2, the gene encoding the p55 accessory subunit of pol γ, that causes progressive external ophthalmoplegia with multiple mtDNA deletions and cytochrome c oxidase (COX)–deficient muscle fibers. Biochemical characterization of purified, recombinant G451E-substituted p55 protein in vitro revealed incomplete stimulation of the catalytic subunit due to compromised subunit interaction. Although G451E p55 retains a wild-type ability to bind DNA, it fails to enhance the DNA-binding strength of the p140-p55 complex. In vivo, the disease most likely arises through haplotype insufficiency or heterodimerization of the mutated and wild-type proteins, which promote mtDNA deletions by stalling the DNA replication fork. The progressive accumulation of mtDNA deletions causes COX deficiency in muscle fibers and results in the clinical phenotype. PMID:16685652

Impact of Ancillary Subunits on Ventricular Repolarization

PubMed Central

Abbott, Geoffrey W.; Xu, Xianghua; Roepke, Torsten K.

2007-01-01

Voltage-gated potassium (Kv) channels generate the outward K+ ion currents that constitute the primary force in ventricular repolarization. Kv channels comprise tetramers of pore-forming α subunits and, in probably the majority of cases in vivo, ancillary or β subunits that help define the properties of the Kv current generated. Ancillary subunits can be broadly categorized as cytoplasmic or transmembrane, and can modify Kv channel trafficking, conductance, gating, ion selectivity, regulation and pharmacology. Because of their often profound effects on Kv channel function, studies of the molecular correlates of ventricular repolarization must take into account ancillary subunits as well as α subunits. Cytoplasmic ancillary subunits include the Kvβ subunits, which regulate a range of Kv channels and may link channel gating to redox potential; and the KChIPs, which appear most often associated with Kv4 subfamily channels that generate the ventricular Ito current. Transmembrane ancillary subunits include the MinK-related proteins (MiRPs) encoded by KCNE genes, which modulate members of most Kv α subunit subfamilies; and the putative 12-transmembrane domain KCR1 protein which modulates hERG. In some cases, such as the ventricular IKs channel complex, it is well-established that the KCNQ1 α subunit must co-assemble with the MinK (KCNE1) single transmembrane domain ancillary subunit for recapitulation of the characteristic, unusually slowly-activating IKs current. In other cases it is not so clear-cut, and in particular the roles of the other MinK-related proteins (MiRPs 1–4) in regulating cardiac Kv channels such as KCNQ1 and hERG in vivo are under debate. MiRP1 alters hERG function and pharmacology, and inherited MiRP1 mutations are associated with inherited and acquired arrhythmias, but controversy exists over the native role of MiRP1 in regulating hERG (and therefore ventricular IKr) in vivo. Some ancillary subunits may exhibit varied expression to shape

12 CFR 367.16 - Ethics Counselor decisions.

Code of Federal Regulations, 2011 CFR

2011-01-01

... 12 Banks and Banking 4 2011-01-01 2011-01-01 false Ethics Counselor decisions. 367.16 Section 367... POLICY SUSPENSION AND EXCLUSION OF CONTRACTOR AND TERMINATION OF CONTRACTS § 367.16 Ethics Counselor... disputed material facts, the Ethics Counselor shall base the decision on the facts as found, together with...

12 CFR 367.16 - Ethics Counselor decisions.

Code of Federal Regulations, 2010 CFR

2010-01-01

... 12 Banks and Banking 4 2010-01-01 2010-01-01 false Ethics Counselor decisions. 367.16 Section 367... POLICY SUSPENSION AND EXCLUSION OF CONTRACTOR AND TERMINATION OF CONTRACTS § 367.16 Ethics Counselor... disputed material facts, the Ethics Counselor shall base the decision on the facts as found, together with...

12 CFR 367.16 - Ethics Counselor decisions.

Code of Federal Regulations, 2013 CFR

2013-01-01

... 12 Banks and Banking 5 2013-01-01 2013-01-01 false Ethics Counselor decisions. 367.16 Section 367... POLICY SUSPENSION AND EXCLUSION OF CONTRACTOR AND TERMINATION OF CONTRACTS § 367.16 Ethics Counselor... disputed material facts, the Ethics Counselor shall base the decision on the facts as found, together with...

12 CFR 367.16 - Ethics Counselor decisions.

Code of Federal Regulations, 2014 CFR

2014-01-01

... 12 Banks and Banking 5 2014-01-01 2014-01-01 false Ethics Counselor decisions. 367.16 Section 367... POLICY SUSPENSION AND EXCLUSION OF CONTRACTOR AND TERMINATION OF CONTRACTS § 367.16 Ethics Counselor... disputed material facts, the Ethics Counselor shall base the decision on the facts as found, together with...

12 CFR 367.16 - Ethics Counselor decisions.

Code of Federal Regulations, 2012 CFR

2012-01-01

... 12 Banks and Banking 5 2012-01-01 2012-01-01 false Ethics Counselor decisions. 367.16 Section 367... POLICY SUSPENSION AND EXCLUSION OF CONTRACTOR AND TERMINATION OF CONTRACTS § 367.16 Ethics Counselor... disputed material facts, the Ethics Counselor shall base the decision on the facts as found, together with...

Role of regulatory subunits and protein kinase inhibitor (PKI) in determining nuclear localization and activity of the catalytic subunit of protein kinase A.

PubMed

Wiley, J C; Wailes, L A; Idzerda, R L; McKnight, G S

1999-03-05

Regulation of protein kinase A by subcellular localization may be critical to target catalytic subunits to specific substrates. We employed epitope-tagged catalytic subunit to correlate subcellular localization and gene-inducing activity in the presence of regulatory subunit or protein kinase inhibitor (PKI). Transiently expressed catalytic subunit distributed throughout the cell and induced gene expression. Co-expression of regulatory subunit or PKI blocked gene induction and prevented nuclear accumulation. A mutant PKI lacking the nuclear export signal blocked gene induction but not nuclear accumulation, demonstrating that nuclear export is not essential to inhibit gene induction. When the catalytic subunit was targeted to the nucleus with a nuclear localization signal, it was not sequestered in the cytoplasm by regulatory subunit, although its activity was completely inhibited. PKI redistributed the nuclear catalytic subunit to the cytoplasm and blocked gene induction, demonstrating that the nuclear export signal of PKI can override a strong nuclear localization signal. With increasing PKI, the export process appeared to saturate, resulting in the return of catalytic subunit to the nucleus. These results demonstrate that both the regulatory subunit and PKI are able to completely inhibit the gene-inducing activity of the catalytic subunit even when the catalytic subunit is forced to concentrate in the nuclear compartment.

Genetic analysis of neuronal ionotropic glutamate receptor subunits

PubMed Central

Granger, Adam J; Gray, John A; Lu, Wei; Nicoll, Roger A

2011-01-01

Abstract In the brain, fast, excitatory synaptic transmission occurs primarily through AMPA- and NMDA-type ionotropic glutamate receptors. These receptors are composed of subunit proteins that determine their biophysical properties and trafficking behaviour. Therefore, determining the function of these subunits and receptor subunit composition is essential for understanding the physiological properties of synaptic transmission. Here, we discuss and evaluate various genetic approaches that have been used to study AMPA and NMDA receptor subunits. These approaches have demonstrated that the GluA1 AMPA receptor subunit is required for activity-dependent trafficking and contributes to basal synaptic transmission, while the GluA2 subunit regulates Ca2+ permeability, homeostasis and trafficking to the synapse under basal conditions. In contrast, the GluN2A and GluN2B NMDA receptor subunits regulate synaptic AMPA receptor content, both during synaptic development and plasticity. Ongoing research in this field is focusing on the molecular interactions and mechanisms that control these functions. To accomplish this, molecular replacement techniques are being used, where native subunits are replaced with receptors containing targeted mutations. In this review, we discuss a single-cell molecular replacement approach which should arguably advance our physiological understanding of ionotropic glutamate receptor subunits, but is generally applicable to study of any neuronal protein. PMID:21768264

Genetic analysis of neuronal ionotropic glutamate receptor subunits.

PubMed

Granger, Adam J; Gray, John A; Lu, Wei; Nicoll, Roger A

2011-09-01

In the brain, fast, excitatory synaptic transmission occurs primarily through AMPA- and NMDA-type ionotropic glutamate receptors. These receptors are composed of subunit proteins that determine their biophysical properties and trafficking behaviour. Therefore, determining the function of these subunits and receptor subunit composition is essential for understanding the physiological properties of synaptic transmission. Here, we discuss and evaluate various genetic approaches that have been used to study AMPA and NMDA receptor subunits. These approaches have demonstrated that the GluA1 AMPA receptor subunit is required for activity-dependent trafficking and contributes to basal synaptic transmission, while the GluA2 subunit regulates Ca(2+) permeability, homeostasis and trafficking to the synapse under basal conditions. In contrast, the GluN2A and GluN2B NMDA receptor subunits regulate synaptic AMPA receptor content, both during synaptic development and plasticity. Ongoing research in this field is focusing on the molecular interactions and mechanisms that control these functions. To accomplish this, molecular replacement techniques are being used, where native subunits are replaced with receptors containing targeted mutations. In this review, we discuss a single-cell molecular replacement approach which should arguably advance our physiological understanding of ionotropic glutamate receptor subunits, but is generally applicable to study of any neuronal protein.

Inherent conformational flexibility of F1-ATPase α-subunit.

PubMed

Hahn-Herrera, Otto; Salcedo, Guillermo; Barril, Xavier; García-Hernández, Enrique

2016-09-01

The core of F1-ATPase consists of three catalytic (β) and three noncatalytic (α) subunits, forming a hexameric ring in alternating positions. A wealth of experimental and theoretical data has provided a detailed picture of the complex role played by catalytic subunits. Although major conformational changes have only been seen in β-subunits, it is clear that α-subunits have to respond to these changes in order to be able to transmit information during the rotary mechanism. However, the conformational behavior of α-subunits has not been explored in detail. Here, we have combined unbiased molecular dynamics (MD) simulations and calorimetrically measured thermodynamic signatures to investigate the conformational flexibility of isolated α-subunits, as a step toward deepening our understanding of its function inside the α3β3 ring. The simulations indicate that the open-to-closed conformational transition of the α-subunit is essentially barrierless, which is ideal to accompany and transmit the movement of the catalytic subunits. Calorimetric measurements of the recombinant α-subunit from Geobacillus kaustophilus indicate that the isolated subunit undergoes no significant conformational changes upon nucleotide binding. Simulations confirm that the nucleotide-free and nucleotide-bound subunits show average conformations similar to that observed in the F1 crystal structure, but they reveal an increased conformational flexibility of the isolated α-subunit upon MgATP binding, which might explain the evolutionary conserved capacity of α-subunits to recognize nucleotides with considerable strength. Furthermore, we elucidate the different dependencies that α- and β-subunits show on Mg(II) for recognizing ATP. Copyright © 2016 Elsevier B.V. All rights reserved.

Polycomb repressive complex 1 modifies transcription of active genes

PubMed Central

Pherson, Michelle; Misulovin, Ziva; Gause, Maria; Mihindukulasuriya, Kathie; Swain, Amanda; Dorsett, Dale

2017-01-01

This study examines the role of Polycomb repressive complex 1 (PRC1) at active genes. The PRC1 and PRC2 complexes are crucial for epigenetic silencing during development of an organism. They are recruited to Polycomb response elements (PREs) and establish silenced domains over several kilobases. Recent studies show that PRC1 is also directly recruited to active genes by the cohesin complex. Cohesin participates broadly in control of gene transcription, but it is unknown whether cohesin-recruited PRC1 also plays a role in transcriptional control of active genes. We address this question using genome-wide RNA sequencing (RNA-seq) and chromatin immunoprecipitation sequencing (ChIP-seq). The results show that PRC1 influences transcription of active genes, and a significant fraction of its effects are likely direct. The roles of different PRC1 subunits can also vary depending on the gene. Depletion of PRC1 subunits by RNA interference alters phosphorylation of RNA polymerase II (Pol II) and occupancy by the Spt5 pausing-elongation factor at most active genes. These effects on Pol II phosphorylation and Spt5 are likely linked to changes in elongation and RNA processing detected by nascent RNA-seq, although the mechanisms remain unresolved. The experiments also reveal that PRC1 facilitates association of Spt5 with enhancers and PREs. Reduced Spt5 levels at these regulatory sequences upon PRC1 depletion coincide with changes in Pol II occupancy and phosphorylation. Our findings indicate that, in addition to its repressive roles in epigenetic gene silencing, PRC1 broadly influences transcription of active genes and may suppress transcription of nonpromoter regulatory sequences. PMID:28782042

Subunit mass analysis for monitoring antibody oxidation.

PubMed

Sokolowska, Izabela; Mo, Jingjie; Dong, Jia; Lewis, Michael J; Hu, Ping

2017-04-01

Methionine oxidation is a common posttranslational modification (PTM) of monoclonal antibodies (mAbs). Oxidation can reduce the in-vivo half-life, efficacy and stability of the product. Peptide mapping is commonly used to monitor the levels of oxidation, but this is a relatively time-consuming method. A high-throughput, automated subunit mass analysis method was developed to monitor antibody methionine oxidation. In this method, samples were treated with IdeS, EndoS and dithiothreitol to generate three individual IgG subunits (light chain, Fd' and single chain Fc). These subunits were analyzed by reversed phase-ultra performance liquid chromatography coupled with an online quadrupole time-of-flight mass spectrometer and the levels of oxidation on each subunit were quantitated based on the deconvoluted mass spectra using the UNIFI software. The oxidation results obtained by subunit mass analysis correlated well with the results obtained by peptide mapping. Method qualification demonstrated that this subunit method had excellent repeatability and intermediate precision. In addition, UNIFI software used in this application allows automated data acquisition and processing, which makes this method suitable for high-throughput process monitoring and product characterization. Finally, subunit mass analysis revealed the different patterns of Fc methionine oxidation induced by chemical and photo stress, which makes it attractive for investigating the root cause of oxidation.

Properties of human brain sodium channel α-subunits expressed in HEK293 cells and their modulation by carbamazepine, phenytoin and lamotrigine

PubMed Central

Qiao, Xin; Sun, Guangchun; Clare, Jeffrey J; Werkman, Taco R; Wadman, Wytse J

2014-01-01

Background and purpose Voltage-activated Na+ channels contain one distinct α-subunit. In the brain NaV1.1, NaV1.2, NaV1.3 and NaV1.6 are the four most abundantly expressed α-subunits. The antiepileptic drugs (AEDs) carbamazepine, phenytoin and lamotrigine have voltage-gated Na+ channels as their primary therapeutic targets. This study provides a systematic comparison of the biophysical properties of these four α-subunits and characterizes their interaction with carbamazepine, phenytoin and lamotrigine. Experimental approach Na+ currents were recorded in voltage-clamp mode in HEK293 cells stably expressing one of the four α-subunits. Key results NaV1.2 and NaV1.3 subunits have a relatively slow recovery from inactivation, compared with the other subunits and NaV1.1 subunits generate the largest window current. Lamotrigine evokes a larger maximal shift of the steady-state inactivation relationship than carbamazepine or phenytoin. Carbamazepine shows the highest binding rate to the α-subunits. Lamotrigine binding to NaV1.1 subunits is faster than to the other α-subunits. Lamotrigine unbinding from the α-subunits is slower than that of carbamazepine and phenytoin. Conclusions and implications The four Na+ channel α-subunits show subtle differences in their biophysical properties, which, in combination with their (sub)cellular expression patterns in the brain, could contribute to differences in neuronal excitability. We also observed differences in the parameters that characterize AED binding to the Na+ channel subunits. Particularly, lamotrigine binding to the four α-subunits suggests a subunit-specific response. Such differences will have consequences for the clinical efficacy of AEDs. Knowledge of the biophysical and binding parameters could be employed to optimize therapeutic strategies and drug development. PMID:24283699

Mediator Complex Subunits MED2, MED5, MED16, and MED23 Genetically Interact in the Regulation of Phenylpropanoid Biosynthesis.

PubMed

Dolan, Whitney L; Dilkes, Brian P; Stout, Jake M; Bonawitz, Nicholas D; Chapple, Clint

2017-12-01

The phenylpropanoid pathway is a major global carbon sink and is important for plant fitness and the engineering of bioenergy feedstocks. In Arabidopsis thaliana , disruption of two subunits of the transcriptional regulatory Mediator complex, MED5a and MED5b, results in an increase in phenylpropanoid accumulation. By contrast, the semidominant MED5b mutation reduced epidermal fluorescence4-3 ( ref4-3 ) results in dwarfism and constitutively repressed phenylpropanoid accumulation. Here, we report the results of a forward genetic screen for suppressors of ref4-3. We identified 13 independent lines that restore growth and/or phenylpropanoid accumulation in the ref4-3 background. Two of the suppressors restore growth without restoring soluble phenylpropanoid accumulation, indicating that the growth and metabolic phenotypes of the ref4-3 mutant can be genetically disentangled. Whole-genome sequencing revealed that all but one of the suppressors carry mutations in MED5b or other Mediator subunits. RNA-seq analysis showed that the ref4-3 mutation causes widespread changes in gene expression, including the upregulation of negative regulators of the phenylpropanoid pathway, and that the suppressors reverse many of these changes. Together, our data highlight the interdependence of individual Mediator subunits and provide greater insight into the transcriptional regulation of phenylpropanoid biosynthesis by the Mediator complex. © 2017 American Society of Plant Biologists. All rights reserved.

ε, a new subunit of RNA polymerase found in gram-positive bacteria.

PubMed

Keller, Andrew N; Yang, Xiao; Wiedermannová, Jana; Delumeau, Olivier; Krásný, Libor; Lewis, Peter J

2014-10-01

RNA polymerase in bacteria is a multisubunit protein complex that is essential for gene expression. We have identified a new subunit of RNA polymerase present in the high-A+T Firmicutes phylum of Gram-positive bacteria and have named it ε. Previously ε had been identified as a small protein (ω1) that copurified with RNA polymerase. We have solved the structure of ε by X-ray crystallography and show that it is not an ω subunit. Rather, ε bears remarkable similarity to the Gp2 family of phage proteins involved in the inhibition of host cell transcription following infection. Deletion of ε shows no phenotype and has no effect on the transcriptional profile of the cell. Determination of the location of ε within the assembly of RNA polymerase core by single-particle analysis suggests that it binds toward the downstream side of the DNA binding cleft. Due to the structural similarity of ε with Gp2 and the fact they bind similar regions of RNA polymerase, we hypothesize that ε may serve a role in protection from phage infection. Copyright © 2014, American Society for Microbiology. All Rights Reserved.

Native and subunit molecular mass and quarternary structure of the hemoglobin from the primitive branchiopod crustacean Triops cancriformis.

PubMed

Rousselot, Morgane; Jaenicke, Elmar; Lamkemeyer, Tobias; Harris, J Robin; Pirow, Ralph

2006-09-01

Many branchiopod crustaceans are endowed with extracellular, high-molecular-weight hemoglobins whose exact structural characteristics have remained a matter of conjecture. By using a broad spectrum of techniques, we provide precise and coherent information on the hemoglobin of one of the phylogenetically 'oldest' extant branchiopods, the tadpole shrimp Triops cancriformis. The hemoglobin dissociated under reducing conditions into two subunits, designated TcHbA and TcHbB, with masses of 35,775+/-4 and 36,055+/-4 Da, respectively, determined by ESI-MS. Nonreducing conditions showed only two disulfide-bridged dimers, a homodimer of TcHbA, designated D1 (71,548+/-5 Da), and the heterodimer D2 (71,828+/-5 Da). Carbamidomethylation of free SH groups revealed the presence of three cysteines per subunit and indicated one intrasubunit and one intersubunit disulfide bridge. Ultracentrifugation and light-scattering experiments under nondenaturating conditions yielded mass estimates that suggested an uneven number of 17 subunits forming the native hemoglobin. This unrealistic number resulted from the presence of two size classes (16-mer and 18-mer), which were recognized by native PAGE and Ferguson plot analysis. ESI-MS revealed three hemoglobin isoforms with masses of 588.1 kDa, 662.0 kDa, and 665.0 kDa. The 16-mer and the smaller 18-mer species are supposed to be composed of TcHbA only, given the dominance of this subunit type in SDS/PAGE. Transmission electron microscopy of negatively stained specimens showed a population of compact molecules with geometrical extensions of 14, 16 and 9 nm. The proposed stoichiometric model of quarternary structure provides the missing link to achieve a mechanistic understanding of the structure-function relationships among the multimeric arthropodan hemoglobins.

Specific Roles of NMDA Receptor Subunits in Mental Disorders.

PubMed

Yamamoto, H; Hagino, Y; Kasai, S; Ikeda, K

2015-01-01

N-methyl-D-aspartate (NMDA) receptor plays important roles in learning and memory. NMDA receptors are a tetramer that consists of two glycine-binding subunits GluN1, two glutamate-binding subunits (i.e., GluN2A, GluN2B, GluN2C, and GluN2D), a combination of a GluN2 subunit and glycine-binding GluN3 subunit (i.e., GluN3A or GluN3B), or two GluN3 subunits. Recent studies revealed that the specific expression and distribution of each subunit are deeply involved in neural excitability, plasticity, and synaptic deficits. The present article summarizes reports on the dysfunction of NMDA receptors and responsible subunits in various neurological and psychiatric disorders, including schizophrenia, autoimmune-induced glutamatergic receptor dysfunction, mood disorders, and autism. A key role for the GluN2D subunit in NMDA receptor antagonist-induced psychosis has been recently revealed.

Anthranilate synthase subunit organization in Chromobacterium violaceum.

PubMed

Carminatti, C A; Oliveira, I L; Recouvreux, D O S; Antônio, R V; Porto, L M

2008-09-16

Tryptophan is an aromatic amino acid used for protein synthesis and cellular growth. Chromobacterium violaceum ATCC 12472 uses two tryptophan molecules to synthesize violacein, a secondary metabolite of pharmacological interest. The genome analysis of this bacterium revealed that the genes trpA-F and pabA-B encode the enzymes of the tryptophan pathway in which the first reaction is the conversion of chorismate to anthranilate by anthranilate synthase (AS), an enzyme complex. In the present study, the organization and structure of AS protein subunits from C. violaceum were analyzed using bioinformatics tools available on the Web. We showed by calculating molecular masses that AS in C. violaceum is composed of alpha (TrpE) and beta (PabA) subunits. This is in agreement with values determined experimentally. Catalytic and regulatory sites of the AS subunits were identified. The TrpE and PabA subunits contribute to the catalytic site while the TrpE subunit is involved in the allosteric site. Protein models for the TrpE and PabA subunits were built by restraint-based homology modeling using AS enzyme, chains A and B, from Salmonella typhimurium (PDB ID 1I1Q).

Subunit stoichiometry of human muscle chloride channels.

PubMed

Fahlke, C; Knittle, T; Gurnett, C A; Campbell, K P; George, A L

1997-01-01

Voltage-gated Cl- channels belonging to the ClC family appear to function as homomultimers, but the number of subunits needed to form a functional channel is controversial. To determine subunit stoichiometry, we constructed dimeric human skeletal muscle Cl- channels in which one subunit was tagged by a mutation (D136G) that causes profound changes in voltage-dependent gating. Sucrose-density gradient centrifugation experiments indicate that both monomeric and dimeric hClC-1 channels in their native configurations exhibit similar sedimentation properties consistent with a multimeric complex having a molecular mass of a dimer. Expression of the heterodimeric channel in a mammalian cell line results in a homogenous population of Cl- channels exhibiting novel gating properties that are best explained by the formation of heteromultimeric channels with an even number of subunits. Heteromultimeric channels were not evident in cells cotransfected with homodimeric WT-WT and D136G-D136G constructs excluding the possibility that functional hClC-1 channels are assembled from more than two subunits. These results demonstrate that the functional hClC-1 unit consists of two subunits.

Soybean glycinin subunits: Characterization of physicochemical and adhesion properties.

PubMed

Mo, Xiaoqun; Zhong, Zhikai; Wang, Donghai; Sun, Xiuzhi

2006-10-04

Soybean proteins have shown great potential for applications as renewable and environmentally friendly adhesives. The objective of this work was to study physicochemical and adhesion properties of soy glycinin subunits. Soybean glycinin was extracted from soybean flour and then fractionated into acidic and basic subunits with an estimated purity of 90 and 85%, respectively. Amino acid composition of glycinin subunits was determined. The high hydrophobic amino acid content is a major contributor to the solubility behavior and water resistance of the basic subunits. Acidic subunits and glycinin had similar solubility profiles, showing more than 80% solubility at pH 2.0-4.0 or 6.5-12.0, whereas basic subunits had considerably lower solubility with the minimum at pH 4.5-8.0. Thermal analysis using a differential scanning calorimeter suggested that basic subunits form new oligomeric structures with higher thermal stability than glycinin but no highly ordered structures present in isolated acidic subunits. The wet strength of basic subunits was 160% more than that of acidic subunits prepared at their respective isoelectric points (pI) and cured at 130 degrees C. Both pH and the curing temperature significantly affected adhesive performance. High-adhesion water resistance was usually observed for adhesives from protein prepared at their pI values and cured at elevated temperatures. Basic subunits are responsible for the water resistance of glycinin and are a good starting material for the development of water-resistant adhesives.

Most neutralizing human monoclonal antibodies target novel epitopes requiring both Lassa virus glycoprotein subunits

PubMed Central

Robinson, James E.; Hastie, Kathryn M.; Cross, Robert W.; Yenni, Rachael E.; Elliott, Deborah H.; Rouelle, Julie A.; Kannadka, Chandrika B.; Smira, Ashley A.; Garry, Courtney E.; Bradley, Benjamin T.; Yu, Haini; Shaffer, Jeffrey G.; Boisen, Matt L.; Hartnett, Jessica N.; Zandonatti, Michelle A.; Rowland, Megan M.; Heinrich, Megan L.; Martínez-Sobrido, Luis; Cheng, Benson; de la Torre, Juan C.; Andersen, Kristian G.; Goba, Augustine; Momoh, Mambu; Fullah, Mohamed; Gbakie, Michael; Kanneh, Lansana; Koroma, Veronica J.; Fonnie, Richard; Jalloh, Simbirie C.; Kargbo, Brima; Vandi, Mohamed A.; Gbetuwa, Momoh; Ikponmwosa, Odia; Asogun, Danny A.; Okokhere, Peter O.; Follarin, Onikepe A.; Schieffelin, John S.; Pitts, Kelly R.; Geisbert, Joan B.; Kulakoski, Peter C.; Wilson, Russell B.; Happi, Christian T.; Sabeti, Pardis C.; Gevao, Sahr M.; Khan, S. Humarr; Grant, Donald S.; Geisbert, Thomas W.; Saphire, Erica Ollmann; Branco, Luis M.; Garry, Robert F.

2016-01-01

Lassa fever is a severe multisystem disease that often has haemorrhagic manifestations. The epitopes of the Lassa virus (LASV) surface glycoproteins recognized by naturally infected human hosts have not been identified or characterized. Here we have cloned 113 human monoclonal antibodies (mAbs) specific for LASV glycoproteins from memory B cells of Lassa fever survivors from West Africa. One-half bind the GP2 fusion subunit, one-fourth recognize the GP1 receptor-binding subunit and the remaining fourth are specific for the assembled glycoprotein complex, requiring both GP1 and GP2 subunits for recognition. Notably, of the 16 mAbs that neutralize LASV, 13 require the assembled glycoprotein complex for binding, while the remaining 3 require GP1 only. Compared with non-neutralizing mAbs, neutralizing mAbs have higher binding affinities and greater divergence from germline progenitors. Some mAbs potently neutralize all four LASV lineages. These insights from LASV human mAb characterization will guide strategies for immunotherapeutic development and vaccine design. PMID:27161536

Analysis of the mechanism of activation of cAMP-dependent protein kinase through the study of mutants of the yeast regulatory subunit.

PubMed

Zaremberg, V; Moreno, S

1996-04-01

Spontaneous mutations in the gene which encodes the regulatory subunit of cAMP-dependent protein kinase (PKA) of Saccharomyces cerevisiae (BCY1) have been isolated previously [Cannon, J. F., Gibbs, J. B. & Tatchell, K. (1986) Genetics 113, 247-264] by selection of ras2::LEU2 revertants that grew on non-fermentable carbon sources. The revertants were placed into groups of increasing severity based on the number of PKA-dependent traits affected [Cannon, J. F., Gitan, R. & Tatchell, K. (1990) J. Biol. Chem. 265, 11897-11904]. In this work the ras2 mutation has been crossed out in each bcy1 allele and the phenotypes of these mutants have been assessed. The order of severity of the mutants in both genetic backgrounds is maintained but the severity of each mutant in the normal background is higher than in the ras2::LEU2 background. Total catalytic-subunit and regulatory-subunit activities were measured in crude extracts of the bcy1 ras2::LEU2 mutants. With one exception (bcy1-6) the calculated regulatory subunit/catalytic subunit ratios of the bcy1 mutants relative to that of wild-type cells were greater than one. The dependence of PKA activity on cAMP was measured in permeabilized cells. The strains show an activity ratio in the absence and presence of cAMP in the range 0.5-1 for Kemptide phosphorylation. Overexpression of the high-affinity cAMP phosphodiesterase gene (PDE2) in the bcy1 ras2::LEU2 strains did not alter their PKA-dependent phenotypes. However, transformants were not observed from the parental ras2::LEU2 strain and the bcy1-6 ras2::LEU2 strain. The results are discussed with respect to a hypothesis for the molecular mechanism of the differential reversal of ras2 phenotypes by the bcy1 alleles. Mutations in the regulatory subunit are predicted to affect the structure of the holoenzyme such that the catalytic subunit is capable of maintaining an active catalytic state, without the need to dissociate from the regulatory subunit.

Transcriptional regulators of Na,K-ATPase subunits

PubMed Central

Li, Zhiqin; Langhans, Sigrid A.

2015-01-01

The Na,K-ATPase classically serves as an ion pump creating an electrochemical gradient across the plasma membrane that is essential for transepithelial transport, nutrient uptake and membrane potential. In addition, Na,K-ATPase also functions as a receptor, a signal transducer and a cell adhesion molecule. With such diverse roles, it is understandable that the Na,K-ATPase subunits, the catalytic α-subunit, the β-subunit and the FXYD proteins, are controlled extensively during development and to accommodate physiological needs. The spatial and temporal expression of Na,K-ATPase is partially regulated at the transcriptional level. Numerous transcription factors, hormones, growth factors, lipids, and extracellular stimuli modulate the transcription of the Na,K-ATPase subunits. Moreover, epigenetic mechanisms also contribute to the regulation of Na,K-ATPase expression. With the ever growing knowledge about diseases associated with the malfunction of Na,K-ATPase, this review aims at summarizing the best-characterized transcription regulators that modulate Na,K-ATPase subunit levels. As abnormal expression of Na,K-ATPase subunits has been observed in many carcinoma, we will also discuss transcription factors that are associated with epithelial-mesenchymal transition, a crucial step in the progression of many tumors to malignant disease. PMID:26579519

Overexpression of α3/α5/β4 nicotinic receptor subunits modifies impulsive-like behavior.

PubMed

Viñals, Xavier; Molas, Susanna; Gallego, Xavier; Fernández-Montes, Rubén D; Robledo, Patricia; Dierssen, Mara; Maldonado, Rafael

2012-05-01

Recent studies have revealed that sequence variants in genes encoding the α3/α5/β4 nicotinic acetylcholine receptor subunits are associated with nicotine dependence. In this study, we evaluated two specific aspects of executive functioning related to drug addiction (impulsivity and working memory) in transgenic mice over expressing α3/α5/β4 nicotinic receptor subunits. Impulsivity and working memory were evaluated in an operant delayed alternation task, where mice must inhibit responding between 2 and 8s in order to receive food reinforcement. Working memory was also evaluated in a spontaneous alternation task in an open field. Transgenic mice showed less impulsive-like behavior than wild-type controls, and this behavioral phenotype was related to the number of copies of the transgene. Thus, transgenic Line 22 (16-28 copies) showed a more pronounced phenotype than Line 30 (4-5 copies). Overexpression of these subunits in Line 22 reduced spontaneous alternation behavior suggesting deficits in working memory processing in this particular paradigm. These results reveal the involvement of α3/α5/β4 nicotinic receptor subunits in working memory and impulsivity, two behavioral traits directly related to the vulnerability to develop nicotine dependence. Copyright © 2011 Elsevier Ireland Ltd. All rights reserved.

29 CFR 511.16 - Reports.

Code of Federal Regulations, 2014 CFR

2014-07-01

... 29 Labor 3 2014-07-01 2014-07-01 false Reports. 511.16 Section 511.16 Labor Regulations Relating... AMERICAN SAMOA § 511.16 Reports. Promptly after receipt of submissions under § 511.15, the committee or subcommittee will resolve the issues before it and prepare a report containing its findings of fact and...

29 CFR 511.16 - Reports.

Code of Federal Regulations, 2011 CFR

2011-07-01

... 29 Labor 3 2011-07-01 2011-07-01 false Reports. 511.16 Section 511.16 Labor Regulations Relating... AMERICAN SAMOA § 511.16 Reports. Promptly after receipt of submissions under § 511.15, the committee or subcommittee will resolve the issues before it and prepare a report containing its findings of fact and...

29 CFR 511.16 - Reports.

Code of Federal Regulations, 2010 CFR

2010-07-01

... 29 Labor 3 2010-07-01 2010-07-01 false Reports. 511.16 Section 511.16 Labor Regulations Relating... AMERICAN SAMOA § 511.16 Reports. Promptly after receipt of submissions under § 511.15, the committee or subcommittee will resolve the issues before it and prepare a report containing its findings of fact and...

The first transmembrane domain (TM1) of β2-subunit binds to the transmembrane domain S1 of α-subunit in BK potassium channels

PubMed Central

Morera, Francisco J.; Alioua, Abderrahmane; Kundu, Pallob; Salazar, Marcelo; Gonzalez, Carlos; Martinez, Agustin D.; Stefani, Enrico; Toro, Ligia; Latorre, Ramon

2012-01-01

The BK channel is one of the most broadly expressed ion channels in mammals. In many tissues, the BK channel pore-forming α-subunit is associated to an auxiliary β-subunit that modulates the voltage- and Ca2+-dependent activation of the channel. Structural components present in β-subunits that are important for the physical association with the α-subunit are yet unknown. Here, we show through co-immunoprecipitation that the intracellular C-terminus, the second transmembrane domain (TM2) and the extracellular loop of the β2-subunit are dispensable for association with the α-subunit pointing transmembrane domain 1 (TM1) as responsible for the interaction. Indeed, the TOXCAT assay for transmembrane protein–protein interactions demonstrated for the first time that TM1 of the β2-subunit physically binds to the transmembrane S1 domain of the α-subunit. PMID:22710124

28 CFR 51.6 - Political subunits.

Code of Federal Regulations, 2010 CFR

2010-07-01

... 28 Judicial Administration 2 2010-07-01 2010-07-01 false Political subunits. 51.6 Section 51.6 Judicial Administration DEPARTMENT OF JUSTICE (CONTINUED) PROCEDURES FOR THE ADMINISTRATION OF SECTION 5 OF THE VOTING RIGHTS ACT OF 1965, AS AMENDED General Provisions § 51.6 Political subunits. All political...

Transcription elongation factors are involved in programming hormone production in pituitary neuroendocrine GH4C1 cells.

PubMed

Fujita, Toshitsugu; Piuz, Isabelle; Schlegel, Werner

2010-05-05

Transcription elongation of many eukaryotic genes is regulated. Two negative transcription elongation factors, 5,6-dichloro-1-beta-D-ribofuranosylbenzimidazole (DRB) sensitivity-inducing factor (DSIF) and negative elongation factor (NELF) are known to stall collaboratively RNA polymerase II promoter proximally. We discovered that DSIF and NELF are linked to hormone expression in rat pituitary GH4C1 cells. When NELF-E, a subunit of NELF or Spt5, a subunit of DSIF was stably knocked-down, prolactin (PRL) expression was increased both at the mRNA and protein levels. In contrast, stable knock-down of only Spt5 abolished growth hormone (GH) expression. Transient NELF-E knock-down increased coincidentally PRL expression and enhanced transcription of a PRL-promoter reporter gene. However, no direct interaction of NELF with the PRL gene could be demonstrated by chromatin immuno-precipitation. Thus, NELF suppressed PRL promoter activity indirectly. In conclusion, transcription regulation by NELF and DSIF is continuously involved in the control of hormone production and may contribute to neuroendocrine cell differentiation. Copyright 2010 Elsevier Ireland Ltd. All rights reserved.

Serological reactivity of patients with Echinococcus infections (E. granulosus, E. vogeli, and E. multilocularis) against three antigen B subunits.

PubMed

de la Rue, Mário L; Yamano, Kimiaki; Almeida, Cybele E; Iesbich, Margarete P; Fernandes, Cloé D; Goto, Akiko; Kouguchi, Hirokazu; Takahashi, Kenichi

2010-02-01

In serodiagnosis of cystic echinococcosis (CE) by Echinococcus granulosus infection, antigen B (AgB) has been utilized worldwide. However, it is known that about 40% of sera with alveolar echinococcosis (AE) by Echinococcus multilocularis infection recognize AgB. Furthermore, cross-reaction against AgB was also reported in sera from polycystic echinococcosis (PE) patients with Echinococcus vogeli infection. These findings indicate that AgB is widely common to the genus Echinococcus. On the other hand, AgB has several subunits, which are composed of the smallest 8-kDa subunit. In this study, reactivities of patient sera with three kinds of Echinococcus infections (CE, PE, and AE) were compared simultaneously under the same condition against three subunits of AgB (8, 16, and 24 kDa). Many articles have referred the fundamental 8- kDa subunit as a diagnostic antigen for CE. However, the reactivity for the 8-kDa subunit of the CE patient was not so high (47.7%) in this study. Furthermore, there are many cases in which serum of patients with PE or AE also recognizes this subunit (66.7% in PE; 45.9% in AE). AgB is effective for the detection of the genus Echinococcus infections, but it does not have high species specificity. Therefore, we need to pay attention to cross-reaction in serodiagnosis of CE in areas where plural species coexist.

Subunit compositions of Arabidopsis RNA polymerases I and III reveal Pol I- and Pol III-specific forms of the AC40 subunit and alternative forms of the C53 subunit

PubMed Central

Ream, Thomas S.; Haag, Jeremy R.; Pontvianne, Frederic; Nicora, Carrie D.; Norbeck, Angela D.; Paša-Tolić, Ljiljana; Pikaard, Craig S.

2015-01-01

Using affinity purification and mass spectrometry, we identified the subunits of Arabidopsis thaliana multisubunit RNA polymerases I and III (abbreviated as Pol I and Pol III), the first analysis of their physical compositions in plants. In all eukaryotes examined to date, AC40 and AC19 subunits are common to Pol I (a.k.a. Pol A) and Pol III (a.k.a. Pol C) and are encoded by single genes. Surprisingly, A. thaliana and related species express two distinct AC40 paralogs, one of which assembles into Pol I and the other of which assembles into Pol III. Changes at eight amino acid positions correlate with the functional divergence of Pol I- and Pol III-specific AC40 paralogs. Two genes encode homologs of the yeast C53 subunit and either protein can assemble into Pol III. By contrast, only one of two potential C17 variants, and one of two potential C31 variants were detected in Pol III. We introduce a new nomenclature system for plant Pol I and Pol III subunits in which the 12 subunits that are structurally and functionally homologous among Pols I through V are assigned equivalent numbers. PMID:25813043

Function and selectivity of bromodomains in anchoring chromatin-modifying complexes to promoter nucleosomes.

PubMed

Hassan, Ahmed H; Prochasson, Philippe; Neely, Kristen E; Galasinski, Scott C; Chandy, Mark; Carrozza, Michael J; Workman, Jerry L

2002-11-01

The functions of the SAGA and SWI/SNF complexes are interrelated and can form stable "epigenetic marks" on promoters in vivo. Here we show that stable promoter occupancy by SWI/SNF and SAGA in the absence of transcription activators requires the bromodomains of the Swi2/Snf2 and Gcn5 subunits, respectively, and nucleosome acetylation. This acetylation can be brought about by either the SAGA or NuA4 HAT complexes. The bromodomain in the Spt7 subunit of SAGA is dispensable for this activity but will anchor SAGA if it is swapped into Gcn5, indicating that specificity of bromodomain function is determined in part by the subunit it occupies. Thus, bromodomains within the catalytic subunits of SAGA and SWI/SNF anchor these complexes to acetylated promoter nucleosomes.

Subunit association of gamma-glutamyltranspeptidase of Escherichia coli K-12.

PubMed

Hashimoto, W; Suzuki, H; Nohara, S; Tachi, H; Yamamoto, K; Kumagai, H

1995-12-01

gamma-Glutamyltranspeptidase [EC 2.3.2.2] of Escherichia coli K-12 consists of one large subunit and one small subunit, which can be separated from each other by high-performance liquid chromatography. Using ion spray mass spectrometry, the masses of the large and the small subunit were determined to be 39,207 and 20,015, respectively. The large subunit exhibited no gamma-glutamyltranspeptidase activity and the small subunit had little enzymatic activity, but a mixture of the two subunits showed partial recovery of the enzymatic activity. The results of native-polyacrylamide gel electrophoresis suggested that they could partially recombine, and that the recombined dimer exhibited enzymatic activity. The gene of gamma-glutamyltranspeptidase encoded a signal peptide, and the large and small subunits in a single open reading frame in that order. Two kinds of plasmid were constructed encoding the signal peptide and either the large or the small subunit. A gamma-glutamyltranspeptidase-less mutant of E. coli K-12 was transformed with each plasmid or with both of them. The strain harboring the plasmid encoding each subunit produced a small amount of the corresponding subunit protein in the periplasmic space but exhibited no enzymatic activity. The strain transformed with both plasmids together exhibited the enzymatic activity, but its specific activity was approximately 3% of that of a strain harboring a plasmid encoding the intact structural gene. These results indicate that a portion of the separated large and small subunits can be reconstituted in vitro and exhibit the enzymatic activity, and that the expressed large and small subunits independently are able to associate in vivo and be folded into an active structure, though the specific activity of the associated subunits was much lower than that of native enzyme. This suggests that the synthesis of gamma-glutamyltranspeptidase in a single precursor polypeptide and subsequent processing are more effective to construct

Probing the proton channels in subunit N of Complex I from Escherichia coli through intra-subunit cross-linking.

PubMed

Tursun, Ablat; Zhu, Shaotong; Vik, Steven B

2016-12-01

Respiratory Complex I appears to have 4 sites for proton translocation, which are coupled to the oxidation of NADH and reduction of coenzyme Q. The proton pathways are thought to be made of offset half-channels that connect to the membrane surfaces, and are connected by a horizontal path through the center of the membrane. In this study of the enzyme from Escherichia coli, subunit N, containing one of the sites, was targeted. Pairs of cysteine residues were introduced into neighboring α-helices along the proposed proton pathways. In an effort to constrain conformational changes that might occur during proton translocation, we attempted to form disulfide bonds or methanethiosulfonate bridges between two engineered cysteine residues. Cysteine modification was inferred by the inability of PEG-maleimide to shift the electrophoretic mobility of subunit N, which will occur upon reaction with free sulfhydryl groups. After the cross-linking treatment, NADH oxidase and NADH-driven proton translocation were measured. Ten different pairs of cysteine residues showed evidence of cross-linking. The most significant loss of enzyme activity was seen for residues near the essential Lys 395. This residue is positioned between the proposed proton half-channel to the periplasm and the horizontal connection through subunit N, and is also near the essential Glu 144 of subunit M. The results suggest important conformational changes in this region for the delivery of protons to the periplasm, or for coupling the actions of subunit N to subunit M. Copyright © 2016 Elsevier B.V. All rights reserved.

The SPT+Herschel+ALMA+Spitzer Legacy Survey: The stellar content of high redshift strongly lensed systems

NASA Astrophysics Data System (ADS)

Vieira, Joaquin; Ashby, Matt; Carlstrom, John; Chapman, Scott; DeBreuck, Carlos; Fassnacht, Chris; Gonzalez, Anthony; Phadke, Kedar; Marrone, Dan; Malkan, Matt; Reuter, Cassie; Rotermund, Kaja; Spilker, Justin; Weiss, Axel

2018-05-01

The South Pole Telescope (SPT) has systematically identified 90 high-redshift strongly gravitationally lensed submillimeter galaxies (SMGs) in a 2500 square-degree cosmological survey of the millimeter (mm) sky. These sources are selected by their extreme mm flux, which is largely independent of redshift and lensing configuration. We are undertaking a comprehensive and systematic followup campaign to use these "cosmic magnifying glasses" to study the infrared background in unprecedented detail, inform the condition of the interstellar medium in starburst galaxies at high redshift, and place limits on dark matter substructure. Here we ask for 115.4 hours of deep Spitzer/IRAC imaging to complete our survey of 90 systems to a uniform depth of 30min integrations at 3.6um and 60min at 4.5um. In our sample of 90 systems, 16 have already been fully observed, 30 have been partially observed, and 44 have not been observed at all. Our immediate goals are to: 1) constrain the specific star formation rates of the background high-redshift submillimeter galaxies by combining these Spitzer observations with our APEX, Herschel, and ALMA data, 2) robustly determine the stellar masses and mass-to-light ratios of all the foreground lensing galaxies in the sample by combining these observations with our VLT and Gemini data, the Dark Energy Survey, and ALMA; and 3) provide complete, deep, and uniform NIR coverage of our entire sample of lensed systems to characterize the environments of high redshift SMGs, maximize the discovery potential for additional spectacular and rare sources, and prepare for JWST. This program will provide the cornerstone data set for two PhD theses: Kedar Phadke at Illinois will lead the analysis of stellar masses for the background SMGs, and Kaja Rotermund at Dalhousie will lead the analysis of stellar masses for the foreground lenses.

Subunit interface dynamics in hexadecameric rubisco.

PubMed

van Lun, Michiel; van der Spoel, David; Andersson, Inger

2011-09-02

Ribulose-1,5-bisphosphate (RuBP) carboxylase/oxygenase (Rubisco) plays an important role in the global carbon cycle as a hub for biomass. Rubisco catalyzes not only the carboxylation of RuBP with carbon dioxide but also a competing oxygenation reaction of RuBP with a negative impact on photosynthetic yield. The functional active site is built from two large (L) subunits that form a dimer. The octameric core of four L(2) dimers is held at each end by a cluster of four small (S) subunits, forming a hexadecamer. Each large subunit contacts more than one S subunit. These interactions exploit the dynamic flexibility of Rubisco, which we address in this study. Here, we describe seven different types of interfaces of hexadecameric Rubisco. We have analyzed these interfaces with respect to the size of the interface area and the number of polar interactions, including salt bridges and hydrogen bonds in a variety of Rubisco enzymes from different organisms and different kingdoms of life, including the Rubisco-like proteins. We have also performed molecular dynamics simulations of Rubisco from Chlamydomonas reinhardtii and mutants thereof. From our computational analyses, we propose structural checkpoints of the S subunit to ensure the functionality and/or assembly of the Rubisco holoenzyme. These checkpoints appear to fine-tune the dynamics of the enzyme in a way that could influence enzyme performance. Copyright © 2011 Elsevier Ltd. All rights reserved.

Geranyl diphosphate synthase large subunit, and methods of use

DOEpatents

Croteau, Rodney B.; Burke, Charles C.; Wildung, Mark R.

2001-10-16

A cDNA encoding geranyl diphosphate synthase large subunit from peppermint has been isolated and sequenced, and the corresponding amino acid sequence has been determined. Replicable recombinant cloning vehicles are provided which code for geranyl diphosphate synthase large subunit). In another aspect, modified host cells are provided that have been transformed, transfected, infected and/or injected with a recombinant cloning vehicle and/or DNA sequence encoding geranyl diphosphate synthase large subunit. In yet another aspect, the present invention provides isolated, recombinant geranyl diphosphate synthase protein comprising an isolated, recombinant geranyl diphosphate synthase large subunit protein and an isolated, recombinant geranyl diphosphate synthase small subunit protein. Thus, systems and methods are provided for the recombinant expression of geranyl diphosphate synthase.

Role for Human Mediator Subunit MED25 in Recruitment of Mediator to Promoters by Endoplasmic Reticulum Stress-responsive Transcription Factor ATF6α*

PubMed Central

Sela, Dotan; Conkright, Juliana J.; Chen, Lu; Gilmore, Joshua; Washburn, Michael P.; Florens, Laurence; Conaway, Ronald C.; Conaway, Joan Weliky

2013-01-01

Transcription factor ATF6α functions as a master regulator of endoplasmic reticulum (ER) stress response genes. In response to ER stress, ATF6α translocates from its site of latency in the ER membrane to the nucleus, where it activates RNA polymerase II transcription of ER stress response genes upon binding sequence-specifically to ER stress response enhancer elements (ERSEs) in their promoter-regulatory regions. In a recent study, we demonstrated that ATF6α activates transcription of ER stress response genes by a mechanism involving recruitment to ERSEs of the multisubunit Mediator and several histone acetyltransferase (HAT) complexes, including Spt-Ada-Gcn5 (SAGA) and Ada-Two-A-containing (ATAC) (Sela, D., Chen, L., Martin-Brown, S., Washburn, M.P., Florens, L., Conaway, J.W., and Conaway, R.C. (2012) J. Biol. Chem. 287, 23035–23045). In this study, we extend our investigation of the mechanism by which ATF6α supports recruitment of Mediator to ER stress response genes. We present findings arguing that Mediator subunit MED25 plays a critical role in this process and identify a MED25 domain that serves as a docking site on Mediator for the ATF6α transcription activation domain. PMID:23864652

Not5-dependent co-translational assembly of Ada2 and Spt20 is essential for functional integrity of SAGA

PubMed Central

Kassem, Sari; Villanyi, Zoltan

2017-01-01

Abstract Acetylation of histones regulates gene expression in eukaryotes. In the yeast Saccharomyces cerevisiae it depends mainly upon the ADA and SAGA histone acetyltransferase complexes for which Gcn5 is the catalytic subunit. Previous screens have determined that global acetylation is reduced in cells lacking subunits of the Ccr4–Not complex, a global regulator of eukaryotic gene expression. In this study we have characterized the functional connection between the Ccr4–Not complex and SAGA. We show that SAGA mRNAs encoding a core set of SAGA subunits are tethered together for co-translational assembly of the encoded proteins. Ccr4–Not subunits bind SAGA mRNAs and promote the co-translational assembly of these subunits. This is needed for integrity of SAGA. In addition, we determine that a glycolytic enzyme, the glyceraldehyde-3-phosphate dehydrogenase Tdh3, a prototypical moonlighting protein, is tethered at this site of Ccr4–Not-dependent co-translational SAGA assembly and functions as a chaperone. PMID:28180299

Performance evaluation of the Russian SPT-100 thruster at NASA LeRC

NASA Technical Reports Server (NTRS)

Sankovic, John M.; Hamley, John A.; Haag, Thomas W.

1994-01-01

Performance measurements of a Russian flight-model SPT-100 thruster were obtained as part of a comprehensive program to evaluate engineering issues pertinent to integration with Western spacecraft. Power processing was provided by a US Government developed laboratory power conditioner. When received the thruster had been subjected to only a few hours of acceptance testing by the manufacturer. Accumulated operating time during this study totalled 148 h and included operation of both cathodes. Cathode flow fraction was controlled both manually and using the flow splitter contained within the supplied xenon flow controller. Data were obtained at current levels ranging from 3 A to 5 A and thruster voltages ranging from 200 V to 300 V. Testing centered on the design power of 1.35 kW with a discharge current of 4.5 A. The effects of facility pressure on thruster operation were examined by varying the pressure via injection of xenon into the vacuum chamber. The facility pressure had a significant effect on thruster performance and stability at the conditions tested. Periods of current instabilities were noted throughout the testing period and became more frequent as testing progressed. Performance during periods of stability agreed with previous data obtained in Russian laboratories.

Quantitative RT-PCR for inhibin/activin subunits: measurements of rat hypothalamic and ovarian inhibin/activin subunit mRNAs during the estrous cycle.

PubMed

Murata, T; Takizawa, T; Funaba, M; Fujimura, H; Murata, E; Takahashi, M; Torii, K

1997-02-01

Inhibins (alpha-beta(A) and alpha-beta(B)) and activins (beta(A)-beta(A), beta(A)-beta(B) and beta(B)-beta(B)) were originally isolated from ovarian follicular fluids as FSH secretion modifiers. Inhibin/activin subunits, alpha, beta(A) and beta(B), are widely distributed in several tissues, including gonads and brain, and inhibins and activins have been reported to be involved in ovarian or hypothalamic functions. In this study, we established and employed a competitive RT-PCR assay system for rat inhibin/activin subunits by capillary electrophoresis to determine rat hypothalamic and ovarian inhibin/activin subunit mRNA levels during the estrous cycle. Linearity of standards for alpha, beta(A), and beta(B) subunit assays were between 0.01-0.3 amol, 0.003-0.09 amol and 0.002-0.02 amol of each fragment DNA as a standard, respectively. Hypothalamic beta(A) subunit mRNA during the estrous morning (1000 h) tended to be increased compared with that of the proestrous evening (1700 h), although they were not significantly different. Ovarian alpha subunit mRNA levels tended to be increased during the proestrous morning (1000 h) and were significantly increased in the proestrous evening (1700 h), compared with diestrus and estrus (P < 0.05). Ovarian beta(A) subunit mRNA was also significantly higher in the proestrous evening, compared with diestrus and estrus (P < 0.05), but in the case of beta(B) subunit mRNA there was no difference among diestrus, proestrus and estrus. We thus established a sensitive competitive RT-PCR system for the measurement of inhibin/activin alpha, beta(A) and beta(B) subunits, and this assay system would be helpful for the study of inhibin/activin action in brain and other tissues where these factors are expressed at low levels.

Subunit compositions of Arabidopsis RNA polymerases I and III reveal Pol I- and Pol III-specific forms of the AC40 subunit and alternative forms of the C53 subunit

DOE PAGES

Ream, Thomas S.; Haag, Jeremy R.; Pontvianne, Frederic; ...

2015-05-02

Using affinity purification and mass spectrometry, we identified the subunits of Arabidopsis thaliana multisubunit RNA Polymerases I and III (abbreviated as Pol I and Pol III), providing the first description of their physical compositions in plants. AC40 and AC19 subunits are typically common to Pol I (a.k.a. Pol A) and Pol III (a.k.a. Pol C) and are encoded by single genes whose mutation, in humans, is a cause of the craniofacial disorder, Treacher-Collins Syndrome. Surprisingly, A. thaliana, and related species, express two distinct AC40 paralogs, one of which assembles into Pol I and the other of which assembles into Polmore » III. Changes at eight amino acid positions correlate with this functional divergence of Pol I and Pol III-specific AC40 paralogs. Two genes encode homologs of the yeast C53 subunit, and either variant can assemble into Pol III. By contrast, only one of two potential C17 variants, and one of two potential C31 variants were detected in Pol III. We introduce a new nomenclature system for plant Pol I and Pol III subunits in which the twelve subunits that are structurally and functionally homologous among Pols I through V are assigned equivalent numbers.« less

Subunit compositions of Arabidopsis RNA polymerases I and III reveal Pol I- and Pol III-specific forms of the AC40 subunit and alternative forms of the C53 subunit

DOE Office of Scientific and Technical Information (OSTI.GOV)

Ream, Thomas S.; Haag, Jeremy R.; Pontvianne, Frederic

Using affinity purification and mass spectrometry, we identified the subunits of Arabidopsis thaliana multisubunit RNA Polymerases I and III (abbreviated as Pol I and Pol III), providing the first description of their physical compositions in plants. AC40 and AC19 subunits are typically common to Pol I (a.k.a. Pol A) and Pol III (a.k.a. Pol C) and are encoded by single genes whose mutation, in humans, is a cause of the craniofacial disorder, Treacher-Collins Syndrome. Surprisingly, A. thaliana, and related species, express two distinct AC40 paralogs, one of which assembles into Pol I and the other of which assembles into Polmore » III. Changes at eight amino acid positions correlate with this functional divergence of Pol I and Pol III-specific AC40 paralogs. Two genes encode homologs of the yeast C53 subunit, and either variant can assemble into Pol III. By contrast, only one of two potential C17 variants, and one of two potential C31 variants were detected in Pol III. We introduce a new nomenclature system for plant Pol I and Pol III subunits in which the twelve subunits that are structurally and functionally homologous among Pols I through V are assigned equivalent numbers.« less

beta'-COP, a novel subunit of coatomer.

PubMed Central

Stenbeck, G; Harter, C; Brecht, A; Herrmann, D; Lottspeich, F; Orci, L; Wieland, F T

1993-01-01

Several lines of evidence favour the hypothesis that intracellular biosynthetic protein transport in eukaryotes is mediated by non-clathrin-coated vesicles (for a review see Rothman and Orci, 1992). The vesicles have been isolated and a set of their surface proteins has been characterized as coat proteins (COPs). These COPs exist in the cytosol as a preformed complex, the coatomer, which was prior to this study known to contain six subunits: four (alpha-, beta-, gamma- and delta-COP) with molecular weights between 160 and 58 kDa, and two additional proteins of approximately 36 and 20 kDa, epsilon- and xi-COP. Here we describe a novel subunit of the coatomer complex, beta'-COP. This subunit occurs in amounts stoichiometric to the established COPs both in the coatomer and in nonclathrin-coated vesicles and shows homology to the beta-subunits of trimeric G proteins. Images PMID:8334999

POB3 is required for both transcription and replication in the yeast Saccharomyces cerevisiae.

PubMed Central

Schlesinger, M B; Formosa, T

2000-01-01

Spt16 and Pob3 form stable heterodimers in Saccharomyces cerevisiae, and homologous proteins have also been purified as complexes from diverse eukaryotes. This conserved factor has been implicated in both transcription and replication and may affect both by altering the characteristics of chromatin. Here we describe the isolation and properties of a set of pob3 mutants and confirm that they have defects in both replication and transcription. Mutation of POB3 caused the Spt(-) phenotype, spt16 and pob3 alleles displayed severe synthetic defects, and elevated levels of Pob3 suppressed some spt16 phenotypes. These results are consistent with previous reports that Spt16 and Pob3 act in a complex that modulates transcription. Additional genetic interactions were observed between pob3 mutations and the genes encoding several DNA replication factors, including POL1, CTF4, DNA2, and CHL12. pob3 alleles caused sensitivity to the ribonucleotide reductase inhibitor hydroxyurea, indicating a defect in a process requiring rapid dNTP synthesis. Mutation of the S phase checkpoint gene MEC1 caused pob3 mutants to lose viability rapidly under restrictive conditions, revealing defects in a process monitored by Mec1. Direct examination of DNA contents by flow cytometry showed that S phase onset and progression were delayed when POB3 was mutated. We conclude that Pob3 is required for normal replication as well as for transcription. PMID:10924459

Detection of a Double Relic in the Torpedo Cluster: SPT-CL J0245-5302

NASA Astrophysics Data System (ADS)

Zheng, Q.; Johnston-Hollitt, M.; Duchesne, S. W.; Li, W. T.

2018-06-01

The Torpedo cluster, SPT-CL J0245-5302 (S0295) is a massive, merging cluster at a redshift of z = 0.300, which exhibits a strikingly similar morphology to the Bullet cluster 1E 0657-55.8 (z = 0.296), including a classic bow shock in the cluster's intra-cluster medium revealed by Chandra X-ray observations. We present Australia Telescope Compact Array data centred at 2.1 GHz and Murchison Widefield Array data at frequencies between 72 MHz and 231 MHz which we use to study the properties of the cluster. We characterise a number of discrete and diffuse radio sources in the cluster, including the detection of two previously unknown radio relics on the cluster periphery. The average spectral index of the diffuse emission between 70 MHz and 3.1 GHz is α =-1.63_{-0.10}^{+0.10} and a radio-derived Mach number for the shock in the west of the cluster is calculated as M = 2.04. The Torpedo cluster is thus a double relic system at moderate redshift.

A Herpesviral Immediate Early Protein Promotes Transcription Elongation of Viral Transcripts.

PubMed

Fox, Hannah L; Dembowski, Jill A; DeLuca, Neal A

2017-06-13

Herpes simplex virus 1 (HSV-1) genes are transcribed by cellular RNA polymerase II (RNA Pol II). While four viral immediate early proteins (ICP4, ICP0, ICP27, and ICP22) function in some capacity in viral transcription, the mechanism by which ICP22 functions remains unclear. We observed that the FACT complex (comprised of SSRP1 and Spt16) was relocalized in infected cells as a function of ICP22. ICP22 was also required for the association of FACT and the transcription elongation factors SPT5 and SPT6 with viral genomes. We further demonstrated that the FACT complex interacts with ICP22 throughout infection. We therefore hypothesized that ICP22 recruits cellular transcription elongation factors to viral genomes for efficient transcription elongation of viral genes. We reevaluated the phenotype of an ICP22 mutant virus by determining the abundance of all viral mRNAs throughout infection by transcriptome sequencing (RNA-seq). The accumulation of almost all viral mRNAs late in infection was reduced compared to the wild type, regardless of kinetic class. Using chromatin immunoprecipitation sequencing (ChIP-seq), we mapped the location of RNA Pol II on viral genes and found that RNA Pol II levels on the bodies of viral genes were reduced in the ICP22 mutant compared to wild-type virus. In contrast, the association of RNA Pol II with transcription start sites in the mutant was not reduced. Taken together, our results indicate that ICP22 plays a role in recruiting elongation factors like the FACT complex to the HSV-1 genome to allow for efficient viral transcription elongation late in viral infection and ultimately infectious virion production. IMPORTANCE HSV-1 interacts with many cellular proteins throughout productive infection. Here, we demonstrate the interaction of a viral protein, ICP22, with a subset of cellular proteins known to be involved in transcription elongation. We determined that ICP22 is required to recruit the FACT complex and other transcription

Transducin β-Subunit Can Interact with Multiple G-Protein γ-Subunits to Enable Light Detection by Rod Photoreceptors.

PubMed

Dexter, Paige M; Lobanova, Ekaterina S; Finkelstein, Stella; Spencer, William J; Skiba, Nikolai P; Arshavsky, Vadim Y

2018-01-01

The heterotrimeric G-protein transducin mediates visual signaling in vertebrate photoreceptor cells. Many aspects of the function of transducin were learned from knock-out mice lacking its individual subunits. Of particular interest is the knockout of its rod-specific γ-subunit (Gγ 1 ). Two studies using independently generated mice documented that this knockout results in a considerable >60-fold reduction in the light sensitivity of affected rods, but provided different interpretations of how the remaining α-subunit (Gα t ) mediates phototransduction without its cognate Gβ 1 γ 1 -subunit partner. One study found that the light sensitivity reduction matched a corresponding reduction in Gα t content in the light-sensing rod outer segments and proposed that Gα t activation is supported by remaining Gβ 1 associating with other Gγ subunits naturally expressed in photoreceptors. In contrast, the second study reported the same light sensitivity loss but a much lower, only approximately sixfold, reduction of Gα t and proposed that the light responses of these rods do not require Gβγ at all. To resolve this controversy and elucidate the mechanism driving visual signaling in Gγ 1 knock-out rods, we analyzed both mouse lines side by side. We first determined that the outer segments of both mice have identical Gα t content, which is reduced ∼65-fold from the wild-type (WT) level. We further demonstrated that the remaining Gβ 1 is present in a complex with endogenous Gγ 2 and Gγ 3 subunits and that these complexes exist in wild-type rods as well. Together, these results argue against the idea that Gα t alone supports light responses of Gγ 1 knock-out rods and suggest that Gβ 1 γ 1 is not unique in its ability to mediate vertebrate phototransduction.

Evaluation of commercial skin prick test solutions for selected occupational allergens.

PubMed

van Kampen, V; de Blay, F; Folletti, I; Kobierski, P; Moscato, G; Olivieri, M; Quirce, S; Sastre, J; Walusiak-Skorupa, J; Kotschy-Lang, N; Müsken, H; Mahler, V; Schliemann, S; Ochmann, U; Sültz, J; Worm, M; Sander, I; Zahradnik, E; Brüning, T; Merget, R; Raulf-Heimsoth, M

2013-01-01

Skin prick testing (SPT) is an important step in the diagnosis of IgE-mediated occupational allergic diseases. The outcome of SPT is related to the quality of allergen extracts. Thus, the aim of the study was to assess different commercially available SPT solutions for selected occupational allergens. SPT was performed in 116 bakers, 47 farmers and 33 subjects exposed to natural rubber latex (NRL), all with work-related allergic symptoms. The SPT solutions from different manufacturers (n = 3-5) for wheat flour, rye flour, soy, cow hair/dander, storage mites (Tyrophagus putrescentiae, Lepidoglyphus destructor, Acarus siro) and NRL were analysed with respect to their protein and antigen contents. SPT was carried out in 16 allergy centres in six European countries using standardized procedures. Specific IgE values were used as the gold standard to calculate the sensitivity and specificity of SPT solutions. The optimal cut-point for each SPT solution was determined by Youden Index. Protein and antigen contents and patterns of the SPT solutions varied remarkably depending on the manufacturer. While SPT solutions for wheat flour and soy reached overall low sensitivities, sensitivities of other tested SPT solutions depended on the manufacturer. As a rule, solutions with higher protein and antigen content showed higher sensitivities and test efficiencies. There is a wide variability of SPT solutions for occupational allergens, and the sensitivity of several solutions is low. Thus, improvement and standardization of SPT solutions for occupational allergens is essential. © 2013 John Wiley & Sons A/S. Published by Blackwell Publishing Ltd.

Heteromeric assembly of P2X subunits

PubMed Central

Saul, Anika; Hausmann, Ralf; Kless, Achim; Nicke, Annette

2013-01-01

Transcripts and/or proteins of P2X receptor (P2XR) subunits have been found in virtually all mammalian tissues. Generally more than one of the seven known P2X subunits have been identified in a given cell type. Six of the seven cloned P2X subunits can efficiently form functional homotrimeric ion channels in recombinant expression systems. This is in contrast to other ligand-gated ion channel families, such as the Cys-loop or glutamate receptors, where homomeric assemblies seem to represent the exception rather than the rule. P2XR mediated responses recorded from native tissues rarely match exactly the biophysical and pharmacological properties of heterologously expressed homomeric P2XRs. Heterotrimerization of P2X subunits is likely to account for this observed diversity. While the existence of heterotrimeric P2X2/3Rs and their role in physiological processes is well established, the composition of most other P2XR heteromers and/or the interplay between distinct trimeric receptor complexes in native tissues is not clear. After a description of P2XR assembly and the structure of the intersubunit ATP-binding site, this review summarizes the distribution of P2XR subunits in selected mammalian cell types and the biochemically and/or functionally characterized heteromeric P2XRs that have been observed upon heterologous co-expression of P2XR subunits. We further provide examples where the postulated heteromeric P2XRs have been suggested to occur in native tissues and an overview of the currently available pharmacological tools that have been used to discriminate between homo- and heteromeric P2XRs. PMID:24391538

Drosophila Vps16A is required for trafficking to lysosomes and biogenesis of pigment granules.

PubMed

Pulipparacharuvil, Suprabha; Akbar, Mohammed Ali; Ray, Sanchali; Sevrioukov, Evgueny A; Haberman, Adam S; Rohrer, Jack; Krämer, Helmut

2005-08-15

Mutations that disrupt trafficking to lysosomes and lysosome-related organelles cause multiple diseases, including Hermansky-Pudlak syndrome. The Drosophila eye is a model system for analyzing such mutations. The eye-color genes carnation and deep orange encode two subunits of the Vps-C protein complex required for endosomal trafficking and pigment-granule biogenesis. Here we demonstrate that dVps16A (CG8454) encodes another Vps-C subunit. Biochemical experiments revealed a specific interaction between the dVps16A C-terminus and the Sec1/Munc18 homolog Carnation but not its closest homolog, dVps33B. Instead, dVps33B interacted with a related protein, dVps16B (CG18112). Deep orange bound both Vps16 homologs. Like a deep orange null mutation, eye-specific RNAi-induced knockdown of dVps16A inhibited lysosomal delivery of internalized ligands and interfered with biogenesis of pigment granules. Ubiquitous knockdown of dVps16A was lethal. Together, these findings demonstrate that Drosophila Vps16A is essential for lysosomal trafficking. Furthermore, metazoans have two types of Vps-C complexes with non-redundant functions.

Subunit stoichiometry of the CNG channel of rod photoreceptors.

PubMed

Weitz, Dietmar; Ficek, Nicole; Kremmer, Elisabeth; Bauer, Paul J; Kaupp, U Benjamin

2002-12-05

Cyclic nucleotide-gated (CNG) channels play a central role in the conversion of sensory stimuli into electrical signals. CNG channels form heterooligomeric complexes built of A and B subunits. Here, we study the subunit stoichiometry of the native rod CNG channel by chemical crosslinking. The apparent molecular weight (M(w)) of each crosslink product was determined by SDS-PAGE, and its composition was analyzed by Western blotting using antibodies specific for the A1 or B1 subunit. The number of crosslink products and their M(w) as well as the immunological identification of A1 and B1 subunits in the crosslink products led us to conclude that the native rod CNG channel is a tetramer composed of three A1 and one B1 subunit. This is an example of violation of symmetry in tetrameric channels.

Rotation of Subunits During Catalysis by Escherichia coli F_1-ATPase

NASA Astrophysics Data System (ADS)

Duncan, Thomas M.; Bulygin, Vladimir V.; Zhou, Yuantai; Hutcheon, Marcus L.; Cross, Richard L.

1995-11-01

During oxidative and photo-phosphorylation, F_0F_1-ATP synthases couple the movement of protons down an electrochemical gradient to the synthesis of ATP. One proposed mechanistic feature that has remained speculative is that this coupling process requires the rotation of subunits within F_0F_1. Guided by a recent, high-resolution structure for bovine F_1 [Abrahams, J. P., Leslie, A. G., Lutter, R. & Walker, J. E. (1994) Nature (London) 370, 621-628], we have developed a critical test for rotation of the central γ subunit relative to the three catalytic β subunits in soluble F_1 from Escherichia coli. In the bovine F_1 structure, a specific point of contact between the γ subunit and one of the three catalytic β subunits includes positioning of the homolog of E. coli γ-subunit C87 (γC87) close to the β-subunit 380DELSEED386 sequence. A βD380C mutation allowed us to induce formation of a specific disulfide bond between β and γC87 in soluble E. coli F_1. Formation of the crosslink inactivated βD380C-F_1, and reduction restored full activity. Using a dissociation/reassembly approach with crosslinked βD380C-F_1, we incorporated radiolabeled β subunits into the two noncrosslinked β-subunit positions of F_1. After reduction of the initial nonradio-active β-γ crosslink, only exposure to conditions for catalytic turnover results in similar reactivities of unlabeled and radiolabeled β subunits with γC87 upon reoxidation. The results demonstrate that γ subunit rotates relative to the β subunits during catalysis.

Localization of yeast RNA polymerase I core subunits by immunoelectron microscopy.

PubMed Central

Klinger, C; Huet, J; Song, D; Petersen, G; Riva, M; Bautz, E K; Sentenac, A; Oudet, P; Schultz, P

1996-01-01

Immunoelectron microscopy was used to determine the spatial organization of the yeast RNA polymerase I core subunits on a three-dimensional model of the enzyme. Images of antibody-labeled enzymes were compared with the native enzyme to determine the localization of the antibody binding site on the surface of the model. Monoclonal antibodies were used as probes to identify the two largest subunits homologous to the bacterial beta and beta' subunits. The epitopes for the two monoclonal antibodies were mapped using subunit-specific phage display libraries, thus allowing a direct correlation of the structural data with functional information on conserved sequence elements. An epitope close to conserved region C of the beta-like subunit is located at the base of the finger-like domain, whereas a sequence between conserved regions C and D of the beta'-like subunit is located in the apical region of the enzyme. Polyclonal antibodies outlined the alpha-like subunit AC40 and subunit AC19 which were found co-localized also in the apical region of the enzyme. The spatial location of the subunits is correlated with their biological activity and the inhibitory effect of the antibodies. Images PMID:8887555

Long-Term Consequences for Teens with Older Sexual Partners. Fact Sheet. Publication #2008-16

ERIC Educational Resources Information Center

Schelar, Erin; Ryan, Suzanne; Manlove, Jennifer

2008-01-01

Existing research on the risks of having sex at a young age with an older partner has focused on poorer reproductive health outcomes in the adolescent years, but scant attention has been paid to the longer-term implications of these relationships. Using survey data from high school students, this "Fact Sheet" presents findings from recent Child…

Genetic analysis of the cytoplasmic dynein subunit families.

PubMed

Pfister, K Kevin; Shah, Paresh R; Hummerich, Holger; Russ, Andreas; Cotton, James; Annuar, Azlina Ahmad; King, Stephen M; Fisher, Elizabeth M C

2006-01-01

Cytoplasmic dyneins, the principal microtubule minus-end-directed motor proteins of the cell, are involved in many essential cellular processes. The major form of this enzyme is a complex of at least six protein subunits, and in mammals all but one of the subunits are encoded by at least two genes. Here we review current knowledge concerning the subunits, their interactions, and their functional roles as derived from biochemical and genetic analyses. We also carried out extensive database searches to look for new genes and to clarify anomalies in the databases. Our analysis documents evolutionary relationships among the dynein subunits of mammals and other model organisms, and sheds new light on the role of this diverse group of proteins, highlighting the existence of two cytoplasmic dynein complexes with distinct cellular roles.

Genetic Analysis of the Cytoplasmic Dynein Subunit Families

PubMed Central

Pfister, K. Kevin; Shah, Paresh R; Hummerich, Holger; Russ, Andreas; Cotton, James; Annuar, Azlina Ahmad; King, Stephen M; Fisher, Elizabeth M. C

2006-01-01

Cytoplasmic dyneins, the principal microtubule minus-end-directed motor proteins of the cell, are involved in many essential cellular processes. The major form of this enzyme is a complex of at least six protein subunits, and in mammals all but one of the subunits are encoded by at least two genes. Here we review current knowledge concerning the subunits, their interactions, and their functional roles as derived from biochemical and genetic analyses. We also carried out extensive database searches to look for new genes and to clarify anomalies in the databases. Our analysis documents evolutionary relationships among the dynein subunits of mammals and other model organisms, and sheds new light on the role of this diverse group of proteins, highlighting the existence of two cytoplasmic dynein complexes with distinct cellular roles. PMID:16440056

Two-subunit DNA escort mechanism and inactive subunit bypass in an ultra-fast ring ATPase

DOE PAGES

Liu, Ninning; Chistol, Gheorghe; Bustamante, Carlos

2015-10-09

SpoIIIE is a homo-hexameric dsDNA translocase responsible for completing chromosome segregation in Bacillus subtilis. Here in this study, we use a single-molecule approach to monitor SpoIIIE translocation when challenged with neutral-backbone DNA and non-hydrolyzable ATP analogs. We show that SpoIIIE makes multiple essential contacts with phosphates on the 5'→3' strand in the direction of translocation. Using DNA constructs with two neutral-backbone segments separated by a single charged base pair, we deduce that SpoIIIE’s step size is 2 bp. Finally, experiments with non-hydrolyzable ATP analogs suggest that SpoIIIE can operate with non-consecutive inactive subunits. We propose a two-subunit escort translocation mechanismmore » that is strict enough to enable SpoIIIE to track one DNA strand, yet sufficiently compliant to permit the motor to bypass inactive subunits without arrest. We speculate that such a flexible mechanism arose for motors that, like SpoIIIE, constitute functional bottlenecks where the inactivation of even a single motor can be lethal for the cell.« less

Two-subunit DNA escort mechanism and inactive subunit bypass in an ultra-fast ring ATPase

DOE PAGES

Liu, Ninning; Chistol, Gheorghe; Bustamante, Carlos

2015-10-09

SpoIIIE is a homo-hexameric dsDNA translocase responsible for completing chromosome segregation in Bacillus subtilis . Here, we use a single-molecule approach to monitor SpoIIIE translocation when challenged with neutral-backbone DNA and non-hydrolyzable ATP analogs. We show that SpoIIIE makes multiple essential contacts with phosphates on the 5'→3' strand in the direction of translocation. Using DNA constructs with two neutral-backbone segments separated by a single charged base pair, we deduce that SpoIIIE’s step size is 2 bp. Finally, experiments with non-hydrolyzable ATP analogs suggest that SpoIIIE can operate with non-consecutive inactive subunits. We propose a two-subunit escort translocation mechanism that ismore » strict enough to enable SpoIIIE to track one DNA strand, yet sufficiently compliant to permit the motor to bypass inactive subunits without arrest. We speculate that such a flexible mechanism arose for motors that, like SpoIIIE, constitute functional bottlenecks where the inactivation of even a single motor can be lethal for the cell.« less

17 CFR 165.16 - No immunity.

Code of Federal Regulations, 2013 CFR

2013-04-01

... 17 Commodity and Securities Exchanges 1 2013-04-01 2013-04-01 false No immunity. 165.16 Section... § 165.16 No immunity. The Commodity Whistleblower Incentives and Protections provisions set forth in... information to the Commission with immunity from prosecution. The fact that an individual may become a...

17 CFR 165.16 - No immunity.

Code of Federal Regulations, 2012 CFR

2012-04-01

... 17 Commodity and Securities Exchanges 1 2012-04-01 2012-04-01 false No immunity. 165.16 Section... § 165.16 No immunity. The Commodity Whistleblower Incentives and Protections provisions set forth in... information to the Commission with immunity from prosecution. The fact that an individual may become a...

17 CFR 165.16 - No immunity.

Code of Federal Regulations, 2014 CFR

2014-04-01

... 17 Commodity and Securities Exchanges 2 2014-04-01 2014-04-01 false No immunity. 165.16 Section...) WHISTLEBLOWER RULES § 165.16 No immunity. The Commodity Whistleblower Incentives and Protections provisions set... provide information to the Commission with immunity from prosecution. The fact that an individual may...

Expression of accessory colonization factor subunit A (ACFA) of Vibrio cholerae and ACFA fused to cholera toxin B subunit in transgenic tomato (Solanum lycopersicum).

PubMed

Sharma, Manoj Kumar; Jani, Dewal; Thungapathra, M; Gautam, J K; Meena, L S; Singh, Yogendra; Ghosh, Amit; Tyagi, Akhilesh Kumar; Sharma, Arun Kumar

2008-05-20

In earlier study from our group, cholera toxin B subunit had been expressed in tomato for developing a plant-based vaccine against cholera. In the present investigation, gene for accessory colonization factor (acf) subunit A, earlier reported to be essential for efficient colonization in the intestine, has been expressed in Escherichia coli as well as tomato plants. Gene encoding for a chimeric protein having a fusion of cholera toxin B subunit and accessory colonization factor A was also expressed in tomato to generate more potent combinatorial antigen. CaMV35S promoter with a duplicated enhancer sequence was used for expression of these genes in tomato. Integration of transgenes into tomato genome was confirmed by PCR and Southern hybridization. Expression of the genes was confirmed at transcript and protein levels. Accessory colonization factor A and cholera toxin B subunit fused to this protein accumulated up to 0.25% and 0.08% of total soluble protein, respectively, in the fruits of transgenic plants. Whereas protein purified from E. coli, in combination with cholera toxin B subunit can be used for development of conventional subunit vaccine, tomato fruits expressing these proteins can be used together with tomato plants expressing cholera toxin B subunit for development of oral vaccine against cholera.

TFIIH Subunit Alterations Causing Xeroderma Pigmentosum and Trichothiodystrophy Specifically Disturb Several Steps during Transcription

PubMed Central

Singh, Amita; Compe, Emanuel; Le May, Nicolas; Egly, Jean-Marc

2015-01-01

Mutations in genes encoding the ERCC3 (XPB), ERCC2 (XPD), and GTF2H5 (p8 or TTD-A) subunits of the transcription and DNA-repair factor TFIIH lead to three autosomal-recessive disorders: xeroderma pigmentosum (XP), XP associated with Cockayne syndrome (XP/CS), and trichothiodystrophy (TTD). Although these diseases were originally associated with defects in DNA repair, transcription deficiencies might be also implicated. By using retinoic acid receptor beta isoform 2 (RARB2) as a model in several cells bearing mutations in genes encoding TFIIH subunits, we observed that (1) the recruitment of the TFIIH complex was altered at the activated RARB2 promoter, (2) TFIIH participated in the recruitment of nucleotide excision repair (NER) factors during transcription in a manner different from that observed during NER, and (3) the different TFIIH variants disturbed transcription by having distinct consequences on post-translational modifications of histones, DNA-break induction, DNA demethylation, and gene-loop formation. The transition from heterochromatin to euchromatin was disrupted depending on the variant, illustrating the fact that TFIIH, by contributing to NER factor recruitment, orchestrates chromatin remodeling. The subtle transcriptional differences found between various TFIIH variants thus participate in the phenotypic variability observed among XP, XP/CS, and TTD individuals. PMID:25620205

Child Care in Sweden. Fact Sheets on Sweden.

ERIC Educational Resources Information Center

Swedish Inst., Stockholm.

This fact sheet outlines Sweden's policies of government-supported child care and parental insurance provisions. Swedish families receive: (1) free maternity and child health care; (2) child allowances for each child of 9,000 krona per year through age 16; (3) up to 450 days of paid parental leave for the birth of a child, with 360 days paid at 90…

Diabetes diagnosis and nutrition facts label use among US adults, 2005-2010.

PubMed

An, Ruopeng

2016-08-01

To assess the role of diabetes diagnosis as a potential teachable moment in nutrition facts label use among US adults. Logistic regression analyses were conducted to examine the relationship between diabetes diagnosis status (diagnosed diabetes, undiagnosed diabetes, diagnosed prediabetes, undiagnosed prediabetes, no diabetes or prediabetes) and self-reported nutrition facts label use, adjusted by individual characteristics and survey design. Study sample came from the National Health and Nutrition Examination Survey 2005-2010 waves. A total of 5110 US adults aged 20 years and older were included in the analyses. Diabetes/prediabetes was identified by fasting plasma glucose and glycated Hb testing. People with diagnosed diabetes/prediabetes were substantially more likely to report nutrition facts label use when making daily food purchase decisions compared with those with undiagnosed diabetes/prediabetes, whereas the prevalence of nutrition facts label use was similar between people with undiagnosed diabetes/prediabetes and those without diabetes/prediabetes. The adjusted prevalence (95 % CI) of any and regular nutrition facts label use was 85·93 (82·91, 88·95) % and 55·60 (50·04, 61·16) % among those with diagnosed diabetes, respectively, in comparison to 71·50 (59·64, 83·37) % and 32·88 (19·11, 46·65) % among those with undiagnosed diabetes. Analogously, the adjusted prevalence (95 % CI) of any and regular nutrition facts label use was 81·16 (75·27, 87·06) % and 45·28 (37·28, 53·29) % among those with diagnosed prediabetes, respectively, in comparison to 72·83 (68·06, 77·59) % and 39·95 (34·02, 45·89) % among those with undiagnosed prediabetes. As a potential teachable moment, diabetes diagnosis may positively impact nutrition facts label use and motivate diabetic patients to manage their condition through making healthier food choices.

Essential arginine in subunit a and aspartate in subunit c of FoF1 ATP synthase: effect of repositioning within helix 4 of subunit a and helix 2 of subunit c.

PubMed

Langemeyer, Lars; Engelbrecht, Siegfried

2007-07-01

FoF1 ATP synthase couples proton flow through the integral membrane portion Fo (ab2c10) to ATP-synthesis in the extrinsic F1-part ((alphabeta)3gammadeltaepsilon) (Escherichia coli nomenclature and stoichiometry). Coupling occurs by mechanical rotation of subunits c10gammaepsilon relative to (alphabeta)3deltaab2. Two residues were found to be essential for proton flow through ab2c10, namely Arg210 in subunit a (aR210) and Asp61 in subunits c (cD61). Their deletion abolishes proton flow, but "horizontal" repositioning, by anchoring them in adjacent transmembrane helices, restores function. Here, we investigated the effects of "vertical" repositioning aR210, cD61, or both by one helical turn towards the N- or C-termini of their original helices. Other than in the horizontal the vertical displacement changes the positions of the side chains within the depth of the membrane. Mutant aR210A/aN214R appeared to be short-circuited in that it supported proton conduction only through EF1-depleted EFo, but not in EFoEF1, nor ATP-driven proton pumping. Mutant cD61N/cM65D grew on succinate, retained the ability to synthesize ATP and supported passive proton conduction but apparently not ATP hydrolysis-driven proton pumping.

Optimal cutoff values of allergen-specific immunoglobulin E to house dust mites and animal dander based on skin-prick test results: Analysis in 16,209 patients with allergic rhinitis.

PubMed

2017-10-25

The most common tests for allergen sensitization in patients with allergic rhinitis are the skin-prick test(SPT) and an in vitro test to detect serum specific immunoglobulin E (sIgE). However, in vitro allergen test results were interpreted dichotomically as positive or negative at a threshold of 0.35kU/L of sIgE, regardless of the patient characteristics or antigen types. The purpose of this study was to determine the cutoff value for sIgE in house-dust mites and animal dander, and to analyze differences in cutoff value according to age and gender. A total of 16,209 patients with more than one allergic rhinitis symptom who underwent both SPT and serum sIgE testing were retrospectively evaluated between March 2008 and May 2012. There were 9374 male (57.8%) and 6835 female (42.2%) patients. The mean age was 31.8 years (range, 2-89 years). The criterion standard for allergen sensitization was defined as a wheal of > 3 mm or an allergen-to-histamine ratio of greater than or equal to 1 in SPT results. The Youden index was used to calculate the cutoff value of sIgE. Cutoff values of sIgE for Dermatophagoides pteronyssinus, Dermatophagoides farinae, cat, and dog were 0.69, 1.16, 0.13, and 0.45 kU/L, respectively. The cutoff value of sIgE changed according to age for D. pteronyssinus and D. farinae but not for cat and dog allergens. When categorizingaccording to age group, the cutoff values of sIgE for D. pteronyssinus and D. farinae had a tendency to decrease with age. There was no significant difference in cutoff value according to gender. The cutoff value for sIgE differed for each antigen and changed with age. Physicians should select the proper cutoff value for sIgE for appropriate criteria according to antigen and patient age rather than using a uniform cutoff value.

Optimal cutoff values of allergen-specific immunoglobulin E to house dust mites and animal dander based on skin-prick test results: Analysis in 16,209 patients with allergic rhinitis.

PubMed

Hong, Sang Duk; Ryu, Gwanghui; Seo, Min Young; Jeong, Jong In; Kim, Hyo Yeol; Chung, Seung-Kyu; Dhong, Hun-Jong

2018-01-25

The most common tests for allergen sensitization in patients with allergic rhinitis are the skin-prick test (SPT) and an in vitro test to detect serum specific immunoglobulin E (sIgE). However, in vitro allergen test results were interpreted dichotomically as positive or negative at a threshold of 0.35 kU/L of sIgE, regardless of the patient characteristics or antigen types. The purpose of this study was to determine the cutoff value for sIgE in house-dust mites and animal dander, and to analyze differences in cutoff value according to age and gender. A total of 16,209 patients with more than one allergic rhinitis symptom who underwent both SPT and serum sIgE testing were retrospectively evaluated between March 2008 and May 2012. There were 9374 male (57.8%) and 6835 female (42.2%) patients. The mean age was 31.8 years (range, 2-89 years). The criterion standard for allergen sensitization was defined as a wheal of >3 mm or an allergen-to-histamine ratio of ≥1 in SPT results. The Youden index was used to calculate the cutoff value of sIgE. Cutoff values of sIgE for Dermatophagoides pteronyssinus, Dermatophagoides farinae, cat, and dog were 0.69, 1.16, 0.13, and 0.45 kU/L, respectively. The cutoff value of sIgE changed according to age for D. pteronyssinus and D. farinae but not for cat and dog allergens. When categorizing according to age group, the cutoff values of sIgE for D. pteronyssinus and D. farinae had a tendency to decrease with age. There was no significant difference in cutoff value according to gender. The cutoff value for sIgE differed for each antigen and changed with age. Physicians should select the proper cutoff value for sIgE for appropriate criteria according to antigen and patient age rather than using a uniform cutoff value.

Differential targeting of Gbetagamma-subunit signaling with small molecules.

PubMed

Bonacci, Tabetha M; Mathews, Jennifer L; Yuan, Chujun; Lehmann, David M; Malik, Sundeep; Wu, Dianqing; Font, Jose L; Bidlack, Jean M; Smrcka, Alan V

2006-04-21

G protein betagamma subunits have potential as a target for therapeutic treatment of a number of diseases. We performed virtual docking of a small-molecule library to a site on Gbetagamma subunits that mediates protein interactions. We hypothesized that differential targeting of this surface could allow for selective modulation of Gbetagamma subunit functions. Several compounds bound to Gbetagamma subunits with affinities from 0.1 to 60 muM and selectively modulated functional Gbetagamma-protein-protein interactions in vitro, chemotactic peptide signaling pathways in HL-60 leukocytes, and opioid receptor-dependent analgesia in vivo. These data demonstrate an approach for modulation of G protein-coupled receptor signaling that may represent an important therapeutic strategy.

Loss of Complex I activity in the Escherichia coli enzyme results from truncating the C-terminus of subunit K, but not from cross-linking it to subunits N or L.

PubMed

Zhu, Shaotong; Canales, Alejandra; Bedair, Mai; Vik, Steven B

2016-06-01

Complex I is a multi-subunit enzyme of the respiratory chain with seven core subunits in its membrane arm (A, H, J, K, L, M, and N). In the enzyme from Escherichia coli the C-terminal ten amino acids of subunit K lie along the lateral helix of subunit L, and contribute to a junction of subunits K, L and N on the cytoplasmic surface. Using double cysteine mutagenesis, the cross-linking of subunit K (R99C) to either subunit L (K581C) or subunit N (T292C) was attempted. A partial yield of cross-linked product had no effect on the activity of the enzyme, or on proton translocation, suggesting that the C-terminus of subunit K has no dynamic role in function. To further elucidate the role of subunit K genetic deletions were constructed at the C-terminus. Upon the serial deletion of the last 4 residues of the C-terminus of subunit K, various results were obtained. Deletion of one amino acid had little effect on the activity of Complex I, but deletions of 2 or more amino acids led to total loss of enzyme activity and diminished levels of subunits L, M, and N in preparations of membrane vesicles. Together these results suggest that while the C-terminus of subunit K has no dynamic role in energy transduction by Complex I, it is vital for the correct assembly of the enzyme.

Detection of the kinematic Sunyaev–Zel'dovich effect with DES Year 1 and SPT

DOE Office of Scientific and Technical Information (OSTI.GOV)

Soergel, B.; Flender, S.; Story, K. T.

Here, we detect the kinematic Sunyaev-Zel'dovich (kSZ) effect with a statistical significance ofmore » $$4.2 \\sigma$$ by combining a cluster catalogue derived from the first year data of the Dark Energy Survey (DES) with CMB temperature maps from the South Pole Telescope Sunyaev-Zel'dovich (SPT-SZ) Survey. This measurement is performed with a differential statistic that isolates the pairwise kSZ signal, providing the first detection of the large-scale, pairwise motion of clusters using redshifts derived from photometric data. By fitting the pairwise kSZ signal to a theoretical template we measure the average central optical depth of the cluster sample, $$\\bar{\\tau}_e = (3.75 \\pm 0.89)\\cdot 10^{-3}$$. We compare the extracted signal to realistic simulations and find good agreement with respect to the signal-to-noise, the constraint on $$\\bar{\\tau}_e$$, and the corresponding gas fraction. High-precision measurements of the pairwise kSZ signal with future data will be able to place constraints on the baryonic physics of galaxy clusters, and could be used to probe gravity on scales $$ \\gtrsim 100$$ Mpc.« less

Detection of the kinematic Sunyaev–Zel'dovich effect with DES Year 1 and SPT

DOE PAGES

Soergel, B.; Flender, S.; Story, K. T.; ...

2016-06-17

Here, we detect the kinematic Sunyaev-Zel'dovich (kSZ) effect with a statistical significance ofmore » $$4.2 \\sigma$$ by combining a cluster catalogue derived from the first year data of the Dark Energy Survey (DES) with CMB temperature maps from the South Pole Telescope Sunyaev-Zel'dovich (SPT-SZ) Survey. This measurement is performed with a differential statistic that isolates the pairwise kSZ signal, providing the first detection of the large-scale, pairwise motion of clusters using redshifts derived from photometric data. By fitting the pairwise kSZ signal to a theoretical template we measure the average central optical depth of the cluster sample, $$\\bar{\\tau}_e = (3.75 \\pm 0.89)\\cdot 10^{-3}$$. We compare the extracted signal to realistic simulations and find good agreement with respect to the signal-to-noise, the constraint on $$\\bar{\\tau}_e$$, and the corresponding gas fraction. High-precision measurements of the pairwise kSZ signal with future data will be able to place constraints on the baryonic physics of galaxy clusters, and could be used to probe gravity on scales $$ \\gtrsim 100$$ Mpc.« less

Catalytic Subunit 1 of Protein Phosphatase 2A Is a Subunit of the STRIPAK Complex and Governs Fungal Sexual Development.

PubMed

Beier, Anna; Teichert, Ines; Krisp, Christoph; Wolters, Dirk A; Kück, Ulrich

2016-06-21

The generation of complex three-dimensional structures is a key developmental step for most eukaryotic organisms. The details of the molecular machinery controlling this step remain to be determined. An excellent model system to study this general process is the generation of three-dimensional fruiting bodies in filamentous fungi like Sordaria macrospora Fruiting body development is controlled by subunits of the highly conserved striatin-interacting phosphatase and kinase (STRIPAK) complex, which has been described in organisms ranging from yeasts to humans. The highly conserved heterotrimeric protein phosphatase PP2A is a subunit of STRIPAK. Here, catalytic subunit 1 of PP2A was functionally characterized. The Δpp2Ac1 strain is sterile, unable to undergo hyphal fusion, and devoid of ascogonial septation. Further, PP2Ac1, together with STRIPAK subunit PRO22, governs vegetative and stress-related growth. We revealed in vitro catalytic activity of wild-type PP2Ac1, and our in vivo analysis showed that inactive PP2Ac1 blocks the complementation of the sterile deletion strain. Tandem affinity purification, followed by mass spectrometry and yeast two-hybrid analysis, verified that PP2Ac1 is a subunit of STRIPAK. Further, these data indicate links between the STRIPAK complex and other developmental signaling pathways, implying the presence of a large interconnected signaling network that controls eukaryotic developmental processes. The insights gained in our study can be transferred to higher eukaryotes and will be important for understanding eukaryotic cellular development in general. The striatin-interacting phosphatase and kinase (STRIPAK) complex is highly conserved from yeasts to humans and is an important regulator of numerous eukaryotic developmental processes, such as cellular signaling and cell development. Although functional insights into the STRIPAK complex are accumulating, the detailed molecular mechanisms of single subunits are only partially understood

INTRINSIC REGULATION OF HEMOGLOBIN EXPRESSION BY VARIABLE SUBUNIT INTERFACE STRENGTHS

PubMed Central

Manning, James M.; Popowicz, Anthony M.; Padovan, Julio C.; Chait, Brian T.; Manning, Lois R.

2012-01-01

SUMMARY The expression of the six types of human hemoglobin subunits over time is currently considered to be regulated mainly by transcription factors that bind to upstream control regions of the gene (the “extrinsic” component of regulation). Here we describe how subunit pairing and further assembly to tetramers in the liganded state is influenced by the affinity of subunits for one another (the “intrinsic” component of regulation). The adult hemoglobin dimers have the strongest subunit interfaces and the embryonic hemoglobins are the weakest with fetal hemoglobins of intermediate strength, corresponding to the temporal order of their expression. These variable subunit binding strengths and the attenuating effects of acetylation contribute to the differences with which these hemoglobin types form functional O2-binding tetramers consistent with gene switching. PMID:22129306

A systematic comparison of all mutations in hereditary sensory neuropathy type I (HSAN I) reveals that the G387A mutation is not disease associated.

PubMed

Hornemann, Thorsten; Penno, Anke; Richard, Stephane; Nicholson, Garth; van Dijk, Fleur S; Rotthier, Annelies; Timmerman, Vincent; von Eckardstein, Arnold

2009-04-01

Hereditary sensory neuropathy type 1 (HSAN I) is an autosomal dominant inherited neurodegenerative disorder of the peripheral nervous system associated with mutations in the SPTLC1 subunit of the serine palmitoyltransferase (SPT). Four missense mutations (C133W, C133Y, V144D and G387A) in SPTLC1 were reported to cause HSAN I. SPT catalyses the condensation of Serine and Palmitoyl-CoA, which is the first and rate-limiting step in the de novo synthesis of ceramides. Earlier studies showed that C133W and C133Y mutants have a reduced activity, whereas the impact of the V144D and G387A mutations on the human enzyme was not tested yet. In this paper, we show that none of the HSAN I mutations interferes with SPT complex formation. We demonstrate that also V144D has a reduced SPT activity, however to a lower extent than C133W and C133Y. In contrast, the G387A mutation showed no influence on SPT activity. Furthermore, the growth phenotype of LY-B cells--a SPTLC1 deficient CHO cell line--could be reversed by expressing either the wild-type SPTLC1 or the G387A mutant, but not the C133W mutant. This indicates that the G387A mutation is most likely not directly associated with HSAN I. These findings were genetically confirmed by the identification of a nuclear HSAN family which showed segregation of the G387A variant as a non-synonymous SNP.

Analysis of the Subunit Stoichiometries in Viral Entry

PubMed Central

Magnus, Carsten; Regoes, Roland R.

2012-01-01

Virions of the Human Immunodeficiency Virus (HIV) infect cells by first attaching with their surface spikes to the CD4 receptor on target cells. This leads to conformational changes in the viral spikes, enabling the virus to engage a coreceptor, commonly CCR5 or CXCR4, and consecutively to insert the fusion peptide into the cellular membrane. Finally, the viral and the cellular membranes fuse. The HIV spike is a trimer consisting of three identical heterodimers composed of the gp120 and gp41 envelope proteins. Each of the gp120 proteins in the trimer is capable of attaching to the CD4 receptor and the coreceptor, and each of the three gp41 units harbors a fusion domain. It is still under debate how many of the envelope subunits within a given trimer have to bind to the CD4 receptors and to the coreceptors, and how many gp41 protein fusion domains are required for fusion. These numbers are referred to as subunit stoichiometries. We present a mathematical framework for estimating these parameters individually by analyzing infectivity assays with pseudotyped viruses. We find that the number of spikes that are engaged in mediating cell entry and the distribution of the spike number play important roles for the estimation of the subunit stoichiometries. Our model framework also shows why it is important to subdivide the question of the number of functional subunits within one trimer into the three different subunit stoichiometries. In a second step, we extend our models to study whether the subunits within one trimer cooperate during receptor binding and fusion. As an example for how our models can be applied, we reanalyze a data set on subunit stoichiometries. We find that two envelope proteins have to engage with CD4-receptors and coreceptors and that two fusion proteins must be revealed within one trimer for viral entry. Our study is motivated by the mechanism of HIV entry but the experimental technique and the model framework can be extended to other viral systems

COP9 signalosome subunit 7 from Arabidopsis interacts with and regulates the small subunit of ribonucleotide reductase (RNR2).

PubMed

Halimi, Yair; Dessau, Moshe; Pollak, Shaul; Ast, Tslil; Erez, Tamir; Livnat-Levanon, Nurit; Karniol, Baruch; Hirsch, Joel A; Chamovitz, Daniel A

2011-09-01

The COP9 Signalosome protein complex (CSN) is a pleiotropic regulator of plant development and contains eight-subunits. Six of these subunits contain the PCI motif which mediates specific protein interactions necessary for the integrity of the complex. COP9 complex subunit 7 (CSN7) contains an N-terminal PCI motif followed by a C-terminal extension which is also necessary for CSN function. A yeast-interaction trap assay identified the small subunit of ribonucelotide reductase (RNR2) from Arabidopsis as interacting with the C-terminal section of CSN7. This interaction was confirmed in planta by both bimolecular fluorescence complementation and immuoprecipitation assays with endogenous proteins. The subcellular localization of RNR2 was primarily nuclear in meristematic regions, and cytoplasmic in adult cells. RNR2 was constitutively nuclear in csn7 mutant seedlings, and was also primarily nuclear in wild type seedlings following exposure to UV-C. These two results correlate with constitutive expression of several DNA-damage response genes in csn7 mutants, and to increased tolerance of csn7 seedlings to UV-C treatment. We propose that the CSN is a negative regulator of RNR activity in Arabidopsis.

[Chymotripsin-like activity and subunit composition of proteasomes in human cancers].

PubMed

Kondakova, I V; Spirina, L V; Koval, V D; Shashova, E E; Choinzonov, E L; Ivanova, E V; Kolomiets, L A; Chernyshova, A L; Slonimskaya, E M; Usynin, E A; Afanasyev, S G

2014-01-01

Activity of the proteasome, polyfunctional enzymatic complex, is known to undergo changes during cancer development. This phenomenon is, probably, caused by the changes in subunit composition of proteasomes. In present work, we studied chymotrypsin-like activity of proteasomes, subunit composition and their association in breast cancer, head and neck squamous cell carcinoma, endometrial cancer, renal cancer, bladder cancer, stomach cancer and colorectal cancer. The increase of proteasome activity was revealed in most cancer tissues compared with adjacent tissues except for the renal cell carcinoma. Changes in proteasome activity in cancer tissues compared with correspondent normal tissues were accompanied by modification of its subunit composition. High proteasome activity was observed in combination with an increased expression of immune subunits and/or proteasome activator PA28, associated with activity of 20S proteasome. In breast cancer, head and neck squamous cell carcinoma, bladder cancer, stomach cancer and colorectal cancer we additionally found higher expression of Rpt6 subunit of 26S proteasome. Correlations between chymotrypsin like proteasome activity and subunit expressions were found in human cancer tissues. In summary, we suggest that proteasome ac- tivation and changes in its subunit composition plays an important role in cancer pathogenesis.

Deciphering the function of the CNGB1b subunit in olfactory CNG channels.

PubMed

Nache, Vasilica; Wongsamitkul, Nisa; Kusch, Jana; Zimmer, Thomas; Schwede, Frank; Benndorf, Klaus

2016-07-11

Olfactory cyclic nucleotide-gated (CNG) ion channels are key players in the signal transduction cascade of olfactory sensory neurons. The second messengers cAMP and cGMP directly activate these channels, generating a depolarizing receptor potential. Olfactory CNG channels are composed of two CNGA2 subunits and two modulatory subunits, CNGA4, and CNGB1b. So far the exact role of the modulatory subunits for channel activation is not fully understood. By measuring ligand binding and channel activation simultaneously, we show that in functional heterotetrameric channels not only the CNGA2 subunits and the CNGA4 subunit but also the CNGB1b subunit binds cyclic nucleotides and, moreover, also alone translates this signal to open the pore. In addition, we show that the CNGB1b subunit is the most sensitive subunit in a heterotetrameric channel to cyclic nucleotides and that it accelerates deactivation to a similar extent as does the CNGA4 subunit. In conclusion, the CNGB1b subunit participates in ligand-gated activation of olfactory CNG channels and, particularly, contributes to rapid termination of odorant signal in an olfactory sensory neuron.

Direct photoaffinity labeling of an allosteric site on subunit protein M1 of mouse ribonucleotide reductase by dTTP.

PubMed Central

Eriksson, S; Caras, I W; Martin, D W

1982-01-01

The protein M1 subunit of ribonucleotide reductase contains at least two allosteric nucleotide binding sites that control the capacity of the enzyme to reduce ribonucleotides to the deoxyribonucleotides required for DNA synthesis. Direct photoaffinity labeling of partially purified protein M1 from mouse T-lymphoma (S49) cells was observed after UV irradiation in the presence of dTTP at 0 degrees C. The relative molar incorporation of nucleotide per subunit was 4-8%. Competition experiments showed that the dTTP was bound to an allosteric domain genetically and kinetically defined as the substrate specificity site of the enzyme. An altered protein M1 isolated from a thymidine-resistant mutant cell line showed significantly decreased photoincorporation of dTTP, consistent with the fact that its CDP reductase activity is resistant to feedback inhibition by dTTP. Specific photolabeling of several other proteins with pyrimidine and purine nucleotides was also found, indicating the general usefulness of direct photoaffinity labeling in the study of enzymes involved in nucleotide and nucleic acid metabolism. Images PMID:7033963

Motor loop dysfunction causes impaired cognitive sequencing in patients suffering from Parkinson's disease.

PubMed

Schönberger, Anna R; Hagelweide, Klara; Pelzer, Esther A; Fink, Gereon R; Schubotz, Ricarda I

2015-10-01

Cognitive impairment in Parkinson's disease (PD) is often attributed to dopamine deficiency in the prefrontal-basal ganglia-thalamo-cortical loops. Although recent studies point to a close interplay between motor and cognitive abilities in PD, the so-called "motor loop" connecting supplementary motor area (SMA) and putamen has been considered solely with regard to the patients' motor impairment. Our study challenges this view by testing patients with the serial prediction task (SPT), a cognitive task that requires participants to predict stimulus sequences and particularly engages premotor sites of the motor loop. We hypothesised that affection of the motor loop causes impaired SPT performance, especially when the internal sequence representation is challenged by suspension of external stimuli. As shown for motor tasks, we further expected this impairment to be compensated by hyperactivity of the lateral premotor cortex (PM). We tested 16 male PD patients ON and OFF dopaminergic medication and 16 male age-matched healthy controls in an functional Magnetic Resonance Imaging study. All subjects performed two versions of the SPT: one with on-going sequences (SPT0), and one with sequences containing non-informative wildcards (SPT+) increasing the demands on mnemonic sequence representation. Patients ON (compared to controls) revealed an impaired performance coming along with hypoactivity of SMA and putamen. Patients OFF compared to ON medication, while showing poorer performance, exhibited a significantly increased PM activity for SPT+ vs. SPT0. Furthermore, patients' performance positively co-varied with PM activity, corroborating a compensatory account. Our data reveal a contribution of the motor loop to cognitive impairment in PD, and suggest a close interplay of SMA and PM beyond motor control. Copyright © 2015 Elsevier Ltd. All rights reserved.

Stiffness of γ subunit of F(1)-ATPase.

PubMed

Okuno, Daichi; Iino, Ryota; Noji, Hiroyuki

2010-11-01

F(1)-ATPase is a molecular motor in which the γ subunit rotates inside the α(3)β(3) ring upon adenosine triphosphate (ATP) hydrolysis. Recent works on single-molecule manipulation of F(1)-ATPase have shown that kinetic parameters such as the on-rate of ATP and the off-rate of adenosine diphosphate (ADP) strongly depend on the rotary angle of the γ subunit (Hirono-Hara et al. 2005; Iko et al. 2009). These findings provide important insight into how individual reaction steps release energy to power F(1) and also have implications regarding ATP synthesis and how reaction steps are reversed upon reverse rotation. An important issue regarding the angular dependence of kinetic parameters is that the angular position of a magnetic bead rotation probe could be larger than the actual position of the γ subunit due to the torsional elasticity of the system. In the present study, we assessed the stiffness of two different portions of F(1) from thermophilic Bacillus PS3: the internal part of the γ subunit embedded in the α(3)β(3) ring, and the complex of the external part of the γ subunit and the α(3)β(3) ring (and streptavidin and magnetic bead), by comparing rotational fluctuations before and after crosslinkage between the rotor and stator. The torsional stiffnesses of the internal and remaining parts were determined to be around 223 and 73 pNnm/radian, respectively. Based on these values, it was estimated that the actual angular position of the internal part of the γ subunit is one-fourth of the magnetic bead position upon stalling using an external magnetic field. The estimated elasticity also partially explains the accommodation of the intrinsic step size mismatch between F(o) and F(1)-ATPase.

The multifaceted subunit interfaces of ionotropic glutamate receptors.

PubMed

Green, Tim; Nayeem, Naushaba

2015-01-01

The past fifteen years has seen a revolution in our understanding of ionotropic glutamate receptor (iGluR) structure, starting with the first view of the ligand binding domain (LBD) published in 1998, and in many ways culminating in the publication of the full-length structure of GluA2 in 2009. These reports have revealed not only the central role played by subunit interfaces in iGluR function, but also myriad binding sites within interfaces for endogenous and exogenous factors. Changes in the conformation of inter-subunit interfaces are central to transmission of ligand gating into pore opening (itself a rearrangement of interfaces), and subsequent closure through desensitization. With the exception of the agonist binding site, which is located entirely within individual subunits, almost all modulatory factors affecting iGluRs appear to bind to sites in subunit interfaces. This review seeks to summarize what we currently understand about the diverse roles interfaces play in iGluR function, and to highlight questions for future research. © 2014 The Authors. The Journal of Physiology © 2014 The Physiological Society.

Loss-of-function mutations and inducible RNAi suppression of Arabidopsis LCB2 genes reveal the critical role of sphingolipids in gametophytic and sporophytic cell viability.

PubMed

Dietrich, Charles R; Han, Gongshe; Chen, Ming; Berg, R Howard; Dunn, Teresa M; Cahoon, Edgar B

2008-04-01

Serine palmitoyltransferase (SPT) catalyzes the first step in sphingolipid biosynthesis, and downregulation of this enzyme provides a means for exploring sphingolipid function in cells. We have previously demonstrated that Arabidopsis SPT requires LCB1 and LCB2 subunits for activity, as is the case in other eukaryotes. In this study, we show that Arabidopsis has two genes (AtLCB2a and AtLCB2b) that encode functional isoforms of the LCB2 subunit. No alterations in sphingolipid content or growth were observed in T-DNA mutants for either gene, but homozygous double mutants were not recoverable, suggesting that these genes are functionally redundant. Reciprocal crosses conducted with Atlcb2a and Atlcb2b mutants indicated that lethality is associated primarily with the inability to transmit the lcb2 null genotype through the haploid pollen. Consistent with this, approximately 50% of the pollen obtained from plants homozygous for a mutation in one gene and heterozygous for a mutation in the second gene arrested during transition from uni-nucleate microspore to bicellular pollen. Ultrastructural analyses revealed that these pollen grains contained aberrant endomembranes and lacked an intine layer. To examine sphingolipid function in sporophytic cells, Arabidopsis lines were generated that allowed inducible RNAi silencing of AtLCB2b in an Atlcb2a mutant background. Studies conducted with these lines demonstrated that sphingolipids are essential throughout plant development, and that lethality resulting from LCB2 silencing in seedlings could be partially rescued by supplying exogenous long-chain bases. Overall, these studies provide insights into the genetic and biochemical properties of SPT and sphingolipid function in Arabidopsis.

TFIIH subunit alterations causing xeroderma pigmentosum and trichothiodystrophy specifically disturb several steps during transcription.

PubMed

Singh, Amita; Compe, Emanuel; Le May, Nicolas; Egly, Jean-Marc

2015-02-05

Mutations in genes encoding the ERCC3 (XPB), ERCC2 (XPD), and GTF2H5 (p8 or TTD-A) subunits of the transcription and DNA-repair factor TFIIH lead to three autosomal-recessive disorders: xeroderma pigmentosum (XP), XP associated with Cockayne syndrome (XP/CS), and trichothiodystrophy (TTD). Although these diseases were originally associated with defects in DNA repair, transcription deficiencies might be also implicated. By using retinoic acid receptor beta isoform 2 (RARB2) as a model in several cells bearing mutations in genes encoding TFIIH subunits, we observed that (1) the recruitment of the TFIIH complex was altered at the activated RARB2 promoter, (2) TFIIH participated in the recruitment of nucleotide excision repair (NER) factors during transcription in a manner different from that observed during NER, and (3) the different TFIIH variants disturbed transcription by having distinct consequences on post-translational modifications of histones, DNA-break induction, DNA demethylation, and gene-loop formation. The transition from heterochromatin to euchromatin was disrupted depending on the variant, illustrating the fact that TFIIH, by contributing to NER factor recruitment, orchestrates chromatin remodeling. The subtle transcriptional differences found between various TFIIH variants thus participate in the phenotypic variability observed among XP, XP/CS, and TTD individuals. Copyright © 2015 The American Society of Human Genetics. Published by Elsevier Inc. All rights reserved.

Painkiller (Oxy, Vike) Facts

MedlinePlus

... That People Abuse Alcohol Facts Bath Salts Facts Cocaine (Coke, Crack) Facts Heroin (Smack, Junk) Facts Marijuana ( ... these pills just like they sell heroin or cocaine. Some people borrow or steal these pills from ...

Genetic ablation of the alpha 6-integrin subunit in Tie1Cre mice enhances tumour angiogenesis.

PubMed

Germain, Mitchel; De Arcangelis, Adèle; Robinson, Stephen D; Baker, Marianne; Tavora, Bernardo; D'Amico, Gabriela; Silva, Rita; Kostourou, Vassiliki; Reynolds, Louise E; Watson, Alan; Jones, J Louise; Georges-Labouesse, Elisabeth; Hodivala-Dilke, Kairbaan

2010-02-01

Laminins are expressed highly in blood vessel basement membranes and have been implicated in angiogenesis. alpha6beta1- and alpha6beta4-integrins are major receptors for laminins in endothelial cells, but the precise role of endothelial alpha6-integrin in tumour angiogenesis is not clear. We show that blood vessels in human invasive ductal carcinoma of the breast have decreased expression of the alpha6-integrin-subunit when compared with normal breast tissue. These data suggest that a decrease in alpha6-integrin-subunit expression in endothelial cells is associated with tumour angiogenesis. To test whether the loss of the endothelial alpha6-integrin subunit affects tumour growth and angiogenesis, we generated alpha6fl/fl-Tie1Cre+ mice and showed that endothelial deletion of alpha6-integrin is sufficient to enhance tumour size and tumour angiogenesis in both murine B16F0 melanoma and Lewis cell lung carcinoma. Mechanistically, endothelial alpha6-integrin deficiency elevated significantly VEGF-mediated angiogenesis both in vivo and ex vivo. In particular, alpha6-integrin-deficient endothelial cells displayed increased levels of VEGF-receptor 2 (VEGFR2) and VEGF-mediated downstream ERK1/2 activation. By developing the first endothelial-specific alpha6-knockout mice, we show that the expression of the alpha6-integrin subunit in endothelial cells acts as a negative regulator of angiogenesis both in vivo and ex vivo. Copyright 2009 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.

Immunochemical analysis of Micrococcus lysodeikticus (luteus) F1-ATPase and its subunits.

PubMed

Urban, C; Salton, M R

1983-08-31

The F1-ATPase from Micrococcus lysodeikticus has been purified to 95% protein homogeneity in this laboratory and as all other bacterial F1S, possesses five distinct subunits with molecular weights ranging from 60 000 to 10 000 (Huberman, M. and Salton, M.R.J. (1979) Biochim. Biophys. Acta 547, 230-240). In this communication, we demonstrate the immunochemical reactivities of antibodies to native and SDS-dissociated subunits with the native and dissociated F1-ATPase and show that: (1) the antibodies generated to the native or SDS-dissociated subunits react with the native molecule; (2) all of the subunits comprising the F1 are antigenically unique as determined by crossed immunoelectrophoresis and the Ouchterlony double-diffusion techniques; (3) antibodies to the SDS-denatured individual delta- and epsilon-subunits can be used to destabilize the interaction of these specific subunits with the rest of the native F1; and (4) all subunit antibodies as well as anti-native F1 were found to inhibit ATPase activity to varying degrees, the strongest inhibition being seen with antibodies to the total F1 and anti-alpha- and anti-beta-subunit antibodies. The interaction of specific subunit antibodies may provide a new and novel way to study further and characterize the catalytic portions of F1-ATPases and in general may offer an additional method for the examination of multimeric proteins.

Differential regulation of thyrotropin subunit apoprotein and carbohydrate biosynthesis by thyroid hormone

DOE Office of Scientific and Technical Information (OSTI.GOV)

Taylor, T.; Weintraub, B.D.

1985-04-01

The regulation of TSH apoprotein and carbohydrate biosynthesis by thyroid hormone was studied by incubating pituitaries from normal and hypothyroid (3 weeks post-thyroidectomy) rats in medium containing (/sup 14/C)alanine and (/sup 3/H) glucosamine. After 6 h, samples were sequentially treated with anti-TSH beta to precipitate TSH and free TSH beta, anti-LH beta to clear the sample of LH and free LH beta, then anti-LH alpha to precipitate free alpha-subunit. Total proteins were acid precipitated. All precipitates were subjected to electrophoresis on sodium dodecyl sulfate-polyacrylamide gels, which were then sliced and assayed by scintillation spectrometry. In hypothyroid pituitaries plus medium, (/supmore » 14/C)alanine incorporation in combined and free beta-subunits was 26 times normal and considerably greater than the 3.4-fold increase seen in total protein; combined and free alpha-subunits showed no specific increase in apoprotein synthesis. (/sup 3/H)Glucosamine incorporation in combined alpha- and beta-subunits in hypothyroid samples was 13 and 21 times normal, respectively, and was greater than the 1.9-fold increase in total protein; free alpha-subunit showed no specific increase in carbohydrate synthesis. The glucosamine to alanine ratio, reflecting relative glycosylation of newly synthesized molecules, was increased in hypothyroidism for combined alpha-subunits, but not for combined beta-subunits, free alpha-subunits, or total proteins. In summary, short term hypothyroidism selectively stimulated TSH beta apoprotein synthesis and carbohydrate synthesis of combined alpha- and beta-subunits. Hypothyroidism also increased the relative glycosylation of combined alpha-subunit. Thus, thyroid hormone deficiency appears to alter the rate-limiting step in TSH assembly (i.e. beta-subunit synthesis) as well as the carbohydrate structure of TSH, which may play important roles in its biological function.« less

Differential Targeting of Gβγ-Subunit Signaling with Small Molecules

NASA Astrophysics Data System (ADS)

Bonacci, Tabetha M.; Mathews, Jennifer L.; Yuan, Chujun; Lehmann, David M.; Malik, Sundeep; Wu, Dianqing; Font, Jose L.; Bidlack, Jean M.; Smrcka, Alan V.

2006-04-01

G protein βγ subunits have potential as a target for therapeutic treatment of a number of diseases. We performed virtual docking of a small-molecule library to a site on Gβγ subunits that mediates protein interactions. We hypothesized that differential targeting of this surface could allow for selective modulation of Gβγ subunit functions. Several compounds bound to Gβγ subunits with affinities from 0.1 to 60 μM and selectively modulated functional Gβγ-protein-protein interactions in vitro, chemotactic peptide signaling pathways in HL-60 leukocytes, and opioid receptor-dependent analgesia in vivo. These data demonstrate an approach for modulation of G protein-coupled receptor signaling that may represent an important therapeutic strategy.

Problem-Based Test: Functional Analysis of Mutant 16S rRNAs

ERIC Educational Resources Information Center

Szeberenyi, Jozsef

2010-01-01

Terms to be familiar with before you start to solve the test: ribosome, ribosomal subunits, antibiotics, point mutation, 16S, 5S, and 23S rRNA, Shine-Dalgarno sequence, mRNA, tRNA, palindrome, hairpin, restriction endonuclease, fMet-tRNA, peptidyl transferase, initiation, elongation, termination of translation, expression plasmid, transformation,…

Fuels planning: science synthesis and integration; social issues fact sheet 16: Prescribed fire and visual quality

Treesearch

Christine Esposito

2006-01-01

Research shows that, while prescribed burning and other fuels treatments can lower visual quality in some situations, they can also improve it in others. This fact sheet reviews the visual aspects of different levels of prescribed burning.Other publications in this series...

Electrophysiology and Beyond: Multiple roles of Na+ channel β subunits in development and disease

PubMed Central

Patino, Gustavo A.; Isom, Lori L.

2010-01-01

Voltage-gated Na+ channel (VGSC) β subunits are not “auxiliary.” These multifunctional molecules not only modulate Na+ current (INa), but also function as cell adhesion molecules (CAMs) – playing roles in aggregation, migration, invasion, neurite outgrowth, and axonal fasciculation. β subunits are integral members of VGSC signaling complexes at nodes of Ranvier, axon initial segments, and cardiac intercalated disks, regulating action potential propagation through critical intermolecular and cell-cell communication events. At least in vitro, many β subunit cell adhesive functions occur both in the presence and absence of pore-forming VGSC α subunits, and in vivo β subunits are expressed in excitable as well as non-excitable cells, thus β subunits may play important functional roles on their own, in the absence of α subunits. VGSC β1 subunits are essential for life and appear to be especially important during brain development. Mutations in β subunit genes result in a variety of human neurological and cardiovascular diseases. Moreover, some cancer cells exhibit alterations in β subunit expression during metastasis. In short, these proteins, originally thought of as merely accessory to α subunits, are critical players in their own right in human health and disease. Here we discuss the role of VGSC β subunits in the nervous system. PMID:20600605

Distinctive interactions of the Arabidopsis homolog of the 30 kD subunit of the cleavage and polyadenylation specificity factor (AtCPSF30) with other polyadenylation factor subunits

USDA-ARS?s Scientific Manuscript database

Background: The Arabidopsis ortholog of the 30 kD subunit of the mammalian Cleavage and Polyadenylation Specificity Factor (AtCPSF30) is an RNA-binding endonuclease that is associated with other Arabidopsis CPSF subunits (orthologs of the 160, 100, and 73 kD subunits of CPSF). In order to better u...

Purification of subunits of Escherichia coli DNA gyrase and reconstitution of enzymatic activity.

PubMed

Higgins, N P; Peebles, C L; Sugino, A; Cozzarelli, N R

1978-04-01

Extensively purified DNA gyrase from Escherichia coli is inhibited by nalidixic acid and by novobiocin. The enzyme is composed of two subunits, A and B, which were purified as separate components. Subunit A is the product of the gene controlling sensitivity to nalidixic acid (nalA) because: (i) the electrophoretic mobility of subunit A in the presence of sodium dodecyl sulfate is identical to that of the 105,000-dalton nalA gene product; (ii) mutants that are resistant to nalidixic acid (nalA(r)) produce a drug-resistant subunit A; and (iii) wild-type subunit A confers drug sensitivity to in vitro synthesis of varphiX174 DNA directed by nalA(r) mutants. Subunit B contains a 95,000-dalton polypeptide and is controlled by the gene specifying sensitivity to novobiocin (cou) because cou(r) mutants produce a novobiocin-resistant subunit B and novobiocin-resitant gyrase is made drug sensitive by wild-type subunit B. Subunits A and B associate, so that gyrase was also purified as a complex containing 105,000- and 95,000-dalton polypeptides. This enzyme and gyrase reconstructed from subunits have the same drug sensitivity, K(m) for ATP, and catalytic properties. The same ratio of subunits gives efficient reconstitution of the reactions intrinsic to DNA gyrase, including catalysis of supercoiling of closed duplex DNA, relaxation of supercoiled DNA in the absence of ATP, and site-specific cleavage of DNA induced by sodium dodecyl sulfate.

Indiana crash facts 1997

DOT National Transportation Integrated Search

1997-01-01

Indiana Crash Facts 1997...is the fifth annual publication : of Indiana Crash Facts. This year significant changes have been made : to the book, including the consolidation of the Alcohol Crash Facts and : the Crash Facts book into one publication. A...

Identification of a small molecule that inhibits herpes simplex virus DNA Polymerase subunit interactions and viral replication.

PubMed

Pilger, Beatrice D; Cui, Can; Coen, Donald M

2004-05-01

The interaction between the catalytic subunit Pol and the processivity subunit UL42 of herpes simplex virus DNA polymerase has been characterized structurally and mutationally and is a potential target for novel antiviral drugs. We developed and validated an assay for small molecules that could disrupt the interaction of UL42 and a Pol-derived peptide and used it to screen approximately 16,000 compounds. Of 37 "hits" identified, four inhibited UL42-stimulated long-chain DNA synthesis by Pol in vitro, of which two exhibited little inhibition of polymerase activity by Pol alone. One of these specifically inhibited the physical interaction of Pol and UL42 and also inhibited viral replication at concentrations below those that caused cytotoxic effects. Thus, a small molecule can inhibit this protein-protein interaction, which provides a starting point for the discovery of new antiviral drugs.

31 CFR 16.17 - Rights of parties.

Code of Federal Regulations, 2012 CFR

2012-07-01

... 31 Money and Finance: Treasury 1 2012-07-01 2012-07-01 false Rights of parties. 16.17 Section 16.17 Money and Finance: Treasury Office of the Secretary of the Treasury REGULATIONS IMPLEMENTING THE...; and (h) Submit written beliefs and proposed findings of fact and conclusions of law after the hearing. ...

31 CFR 16.17 - Rights of parties.

Code of Federal Regulations, 2013 CFR

2013-07-01

... 31 Money and Finance: Treasury 1 2013-07-01 2013-07-01 false Rights of parties. 16.17 Section 16.17 Money and Finance: Treasury Office of the Secretary of the Treasury REGULATIONS IMPLEMENTING THE...; and (h) Submit written beliefs and proposed findings of fact and conclusions of law after the hearing. ...

31 CFR 16.17 - Rights of parties.

Code of Federal Regulations, 2010 CFR

2010-07-01

... 31 Money and Finance: Treasury 1 2010-07-01 2010-07-01 false Rights of parties. 16.17 Section 16.17 Money and Finance: Treasury Office of the Secretary of the Treasury REGULATIONS IMPLEMENTING THE...; and (h) Submit written beliefs and proposed findings of fact and conclusions of law after the hearing. ...

31 CFR 16.17 - Rights of parties.

Code of Federal Regulations, 2011 CFR

2011-07-01

... 31 Money and Finance: Treasury 1 2011-07-01 2011-07-01 false Rights of parties. 16.17 Section 16.17 Money and Finance: Treasury Office of the Secretary of the Treasury REGULATIONS IMPLEMENTING THE...; and (h) Submit written beliefs and proposed findings of fact and conclusions of law after the hearing. ...

31 CFR 16.17 - Rights of parties.

Code of Federal Regulations, 2014 CFR

2014-07-01

... 31 Money and Finance: Treasury 1 2014-07-01 2014-07-01 false Rights of parties. 16.17 Section 16.17 Money and Finance: Treasury Office of the Secretary of the Treasury REGULATIONS IMPLEMENTING THE...; and (h) Submit written beliefs and proposed findings of fact and conclusions of law after the hearing. ...

Research Facts on Homeschooling. General Facts and Trends

ERIC Educational Resources Information Center

Ray, Brian D.

2006-01-01

This fact sheet presents the following general facts and trends. Homeschooling may be the fastest-growing form of education in the United States and it is also growing around the world in many nations. There are about 2 million homeschool students in the United States. There were an estimated 1.9 to 2.4 million children (in grades K to 12) home…

Subunits of the Snf1 kinase heterotrimer show interdependence for association and activity.

PubMed

Elbing, Karin; Rubenstein, Eric M; McCartney, Rhonda R; Schmidt, Martin C

2006-09-08

The Snf1 kinase and its mammalian orthologue, the AMP-activated protein kinase (AMPK), function as heterotrimers composed of a catalytic alpha-subunit and two non-catalytic subunits, beta and gamma. The beta-subunit is thought to hold the complex together and control subcellular localization whereas the gamma-subunit plays a regulatory role by binding to and blocking the function of an auto-inhibitory domain (AID) present in the alpha-subunit. In addition, catalytic activity requires phosphorylation by a distinct upstream kinase. In yeast, any one of three Snf1-activating kinases, Sak1, Tos3, or Elm1, can fulfill this role. We have previously shown that Sak1 is the only Snf1-activating kinase that forms a stable complex with Snf1. Here we show that the formation of the Sak1.Snf1 complex requires the beta- and gamma-subunits in vivo. However, formation of the Sak1.Snf1 complex is not necessary for glucose-regulated phosphorylation of the Snf1 activation loop. Snf1 kinase purified from cells lacking the beta-subunits do not contain any gamma-subunit, indicating that the Snf1 kinase does not form a stable alphagamma dimer in vivo. In vitro kinase assays using purified full-length and truncated Snf1 proteins demonstrate that the kinase domain, which lacks the AID, is significantly more active than the full-length Snf1 protein. Addition of purified beta- and gamma-subunits could stimulate the kinase activity of the full-length alpha-subunit but only when all three subunits were present, suggesting an interdependence of all three subunits for assembly of a functional complex.

The gamma subunit of transducin is farnesylated.

PubMed Central

Lai, R K; Perez-Sala, D; Cañada, F J; Rando, R R

1990-01-01

Protein prenylation with farnesyl or geranylgeranyl moieties is an important posttranslational modification that affects the activity of such diverse proteins as the nuclear lamins, the yeast mating factor mata, and the ras oncogene products. In this article, we show that whole retinal cultures incorporate radioactive mevalonic acid into proteins of 23-26 kDa and one of 8 kDa. The former proteins are probably the "small" guanine nucleotide-binding regulatory proteins (G proteins) and the 8-kDa protein is the gamma subunit of the well-studied retinal heterotrimeric G protein (transducin). After deprenylating purified transducin and its subunits with Raney nickel or methyl iodide/base, the adducted prenyl group can be identified as an all-trans-farnesyl moiety covalently linked to a cysteine residue. Thus far, prenylation reactions have been found to occur at cysteine in a carboxyl-terminal consensus CAAX sequence, where C is the cysteine, A is an aliphatic amino acid, and X is undefined. Both the alpha and gamma subunits of transducin have this consensus sequence, but only the gamma subunit is prenylated. Therefore, the CAAX motif is not necessary and sufficient to direct prenylation. Finally, since transducin is the best understood G protein, both structurally and mechanistically, the discovery that it is farnesylated should allow for a quantitative understanding of this post-translational modification. Images PMID:2217200

Molecular mechanics of 30S subunit head rotation.

PubMed

Mohan, Srividya; Donohue, John Paul; Noller, Harry F

2014-09-16

During ribosomal translocation, a process central to the elongation phase of protein synthesis, movement of mRNA and tRNAs requires large-scale rotation of the head domain of the small (30S) subunit of the ribosome. It has generally been accepted that the head rotates by pivoting around the neck helix (h28) of 16S rRNA, its sole covalent connection to the body domain. Surprisingly, we observe that the calculated axis of rotation does not coincide with the neck. Instead, comparative structure analysis across 55 ribosome structures shows that 30S head movement results from flexing at two hinge points lying within conserved elements of 16S rRNA. Hinge 1, although located within the neck, moves by straightening of the kinked helix h28 at the point of contact with the mRNA. Hinge 2 lies within a three-way helix junction that extends to the body through a second, noncovalent connection; its movement results from flexing between helices h34 and h35 in a plane orthogonal to the movement of hinge 1. Concerted movement at these two hinges accounts for the observed magnitudes of head rotation. Our findings also explain the mode of action of spectinomycin, an antibiotic that blocks translocation by binding to hinge 2.

Molecular mechanics of 30S subunit head rotation

PubMed Central

Mohan, Srividya; Donohue, John Paul; Noller, Harry F.

2014-01-01

During ribosomal translocation, a process central to the elongation phase of protein synthesis, movement of mRNA and tRNAs requires large-scale rotation of the head domain of the small (30S) subunit of the ribosome. It has generally been accepted that the head rotates by pivoting around the neck helix (h28) of 16S rRNA, its sole covalent connection to the body domain. Surprisingly, we observe that the calculated axis of rotation does not coincide with the neck. Instead, comparative structure analysis across 55 ribosome structures shows that 30S head movement results from flexing at two hinge points lying within conserved elements of 16S rRNA. Hinge 1, although located within the neck, moves by straightening of the kinked helix h28 at the point of contact with the mRNA. Hinge 2 lies within a three-way helix junction that extends to the body through a second, noncovalent connection; its movement results from flexing between helices h34 and h35 in a plane orthogonal to the movement of hinge 1. Concerted movement at these two hinges accounts for the observed magnitudes of head rotation. Our findings also explain the mode of action of spectinomycin, an antibiotic that blocks translocation by binding to hinge 2. PMID:25187561

16 CFR 1145.16 - Lighters that are intended for igniting smoking materials and that can be operated by children...

Code of Federal Regulations, 2011 CFR

2011-01-01

... smoking materials and that can be operated by children; risks of death or injury. 1145.16 Section 1145.16... for igniting smoking materials and that can be operated by children; risks of death or injury. (a) The... risks of injury associated with the fact that lighters intended for igniting smoking materials can be...

16 CFR 1145.16 - Lighters that are intended for igniting smoking materials and that can be operated by children...

Code of Federal Regulations, 2010 CFR

2010-01-01

... smoking materials and that can be operated by children; risks of death or injury. 1145.16 Section 1145.16... for igniting smoking materials and that can be operated by children; risks of death or injury. (a) The... risks of injury associated with the fact that lighters intended for igniting smoking materials can be...

16 CFR 1145.16 - Lighters that are intended for igniting smoking materials and that can be operated by children...

Code of Federal Regulations, 2012 CFR

2012-01-01

... smoking materials and that can be operated by children; risks of death or injury. 1145.16 Section 1145.16... for igniting smoking materials and that can be operated by children; risks of death or injury. (a) The... risks of injury associated with the fact that lighters intended for igniting smoking materials can be...

16 CFR 1145.16 - Lighters that are intended for igniting smoking materials and that can be operated by children...

Code of Federal Regulations, 2014 CFR

2014-01-01

... smoking materials and that can be operated by children; risks of death or injury. 1145.16 Section 1145.16... for igniting smoking materials and that can be operated by children; risks of death or injury. (a) The... risks of injury associated with the fact that lighters intended for igniting smoking materials can be...

The H,K-ATPase beta-subunit can act as a surrogate for the beta-subunit of Na,K-pumps.

PubMed

Horisberger, J D; Jaunin, P; Reuben, M A; Lasater, L S; Chow, D C; Forte, J G; Sachs, G; Rossier, B C; Geering, K

1991-10-15

Na,K-ATPase and H,K-ATPase are the only members of the P-type ATPases in which a glycosylated beta-subunit is part of the purified active enzyme. In this study, we have followed the synthesis and the posttranslational processing of the beta-subunit of H,K-ATPase (beta HK) in Xenopus oocytes injected with beta HK cRNA and have tested whether it can act as a surrogate for the beta-subunit of Na,K-ATPase (beta NaK) to support the functional expression of Na,K-pumps. In Xenopus oocytes, beta HK is processed from an Endo H-sensitive 51-kDa coreglycosylated form to an Endo H-resistant 71-kDa fully glycosylated form. Similar to beta NaK, beta HK can stabilize and increase the trypsin resistance of alpha-subunits of Na,K-ATPase (alpha NaK). Finally, expression of beta HK together with alpha NaK leads to an increased number of ouabain binding sites at the plasma membrane accompanied by an increased Rb+ uptake and Na,K-pump current. Our data suggest that beta HK, similar to beta NaK, can assemble to alpha NaK, support the structural maturation and the intracellular transport of catalytic alpha NaK, and ultimately form active alpha NaK-beta HK complexes with Na,K-pump transport properties.

The Development and Institutionalization of Subunit Power in Organizations.

ERIC Educational Resources Information Center

Boeker, Warren

1989-01-01

Examines the effects of founding events on the evolution of subunit importance in the semiconductor industry from 1958 to 1985. Distributions of power and subunit importance represent not only influences of current conditions, but also vestiges of earlier events, including the institution's founding. Includes 55 references. (MLH)

Yeast Inner-Subunit PA-NZ-1 Labeling Strategy for Accurate Subunit Identification in a Macromolecular Complex through Cryo-EM Analysis.

PubMed

Wang, Huping; Han, Wenyu; Takagi, Junichi; Cong, Yao

2018-05-11

Cryo-electron microscopy (cryo-EM) has been established as one of the central tools in the structural study of macromolecular complexes. Although intermediate- or low-resolution structural information through negative staining or cryo-EM analysis remains highly valuable, we lack general and efficient ways to achieve unambiguous subunit identification in these applications. Here, we took advantage of the extremely high affinity between a dodecapeptide "PA" tag and the NZ-1 antibody Fab fragment to develop an efficient "yeast inner-subunit PA-NZ-1 labeling" strategy that when combined with cryo-EM could precisely identify subunits in macromolecular complexes. Using this strategy combined with cryo-EM 3D reconstruction, we were able to visualize the characteristic NZ-1 Fab density attached to the PA tag inserted into a surface-exposed loop in the middle of the sequence of CCT6 subunit present in the Saccharomyces cerevisiae group II chaperonin TRiC/CCT. This procedure facilitated the unambiguous localization of CCT6 in the TRiC complex. The PA tag was designed to contain only 12 amino acids and a tight turn configuration; when inserted into a loop, it usually has a high chance of maintaining the epitope structure and low likelihood of perturbing the native structure and function of the target protein compared to other tagging systems. We also found that the association between PA and NZ-1 can sustain the cryo freezing conditions, resulting in very high occupancy of the Fab in the final cryo-EM images. Our study demonstrated the robustness of this strategy combined with cryo-EM in efficient and accurate subunit identification in challenging multi-component complexes. Copyright © 2018 Elsevier Ltd. All rights reserved.

Human thyrotropin receptor subunits characterized by thyrotropin affinity purification and western blotting.

PubMed

Leedman, P J; Newman, J D; Harrison, L C

1989-07-01

We studied the subunit structure of the human TSH receptor in thyroid tissue from patients with Graves' disease and multinodular goiter by TSH affinity chromatography, immunoprecipitation with Graves' immunoglobulins (Igs), and a modified technique of Western blotting. Human TSH receptor-binding activity was purified about 1,270-fold by sequential affinity chromatography on wheat germ lectin-agarose and TSH-agarose. Analysis by sodium dodecyl sulfate-polyacrylamide gel electrophoresis of nonreduced affinity-purified receptors eluted in sodium dodecyl sulfate sample buffer revealed three noncovalently linked subunits of 70,000, 50,000, and 35,000 mol wt. When reduced, a major subunit of 25,000 mol wt was identified. When 3 mol/L NaCl was used to elute affinity-purified receptors only the 50,000 mol wt nonreduced subunit was detected. This subunit bound [125I]bovine TSH and was precipitated by Graves' Igs. Modifications to the conventional Western blotting technique enabled thyroglobulin components (approximately 220,000 mol wt), thyroid microsomal antigen (a doublet of approximately 110,000 mol wt), and putative TSH receptor subunits of 70,000 and 50,000 mol wt to be identified in thyroid particulate membranes by Graves' Igs. Blotting of affinity-purified receptors eluted in sodium dodecyl sulfate sample buffer revealed subunits of either 70,000 or 50,000 mol wt, with a minority of Graves' serum samples. We conclude that the nonreduced human TSH receptor is an oligomeric complex comprising three different subunits of 70,000, 50,000, and 35,000 mol wt. The reduced receptor exists as a single subunit of 25,000 mol wt, which may be disulfide linked to form the higher mol wt forms. The 70,000 and 50,000 mol wt subunits contain epitopes that bind Graves' Igs in modified Western blots, thus directly confirming that the human TSH receptor is a target for Graves' Igs.

Topographic antigenic determinants recognized by monoclonal antibodies on human choriogonadotropin beta-subunit

DOE Office of Scientific and Technical Information (OSTI.GOV)

Bidart, J.M.; Troalen, F.; Salesse, R.

1987-06-25

We describe a first attempt to study the antibody-combining sites recognized by monoclonal antibodies raised against the beta-subunit of human choriogonadotropin (hCG). Two groups of antibodies were first defined by their ability to recognize only the free beta-subunit or the free and combined subunit. Antibodies FBT-11 and FBT-11-L bind only to hCG beta-subunit but not to hCG, whereas antibodies FBT-10 and D1E8 bind to both the beta-subunit and the hormone. In both cases, the antigenic determinants were localized to the core of the protein (residues 1-112), indicating the weak immunogenicity of the specific carboxyl-terminal extension of hCG-beta. Nine synthetic peptidesmore » spanning different regions of hCG-beta and lutropin-beta were assessed for their capacity to inhibit antibody binding. A synthetic peptide inclusive of the NH2-terminal region (residues 1-7) of the hCG beta-subunit was found to inhibit binding to the radiolabeled subunit of a monoclonal antibody specific for free hCG-beta (FBT-11). Further delineation of the antigenic site recognized by this antibody provided evidence for the involvement of fragment 82-92. Moreover, monoclonal antibody FBT-11 inhibited the recombination of hCG-beta to hCG-alpha, indicating that its antigenic determinant might be located nearby or in the hCG-beta portion interacting with the alpha-subunit. Binding of monoclonal antibody FBT-10, corresponding to the second antigenic determinant, was weakly inhibited by fragment 82-105 and did not impair the recombination of the hCG beta-subunit to the hCG alpha-subunit. Its combining site appeared to be located in a region of the intact native choriogonadotropin present at the surface of the hormone-receptor complex.« less

The Ror receptor tyrosine kinase CAM-1 is required for ACR-16-mediated synaptic transmission at the C. elegans neuromuscular junction.

PubMed

Francis, Michael M; Evans, Susan P; Jensen, Michael; Madsen, David M; Mancuso, Joel; Norman, Kenneth R; Maricq, Andres Villu

2005-05-19

Nicotinic (cholinergic) neurotransmission plays a critical role in the vertebrate nervous system, underlies nicotine addiction, and nicotinic receptor dysfunction leads to neurological disorders. The C. elegans neuromuscular junction (NMJ) shares many characteristics with neuronal synapses, including multiple classes of postsynaptic currents. Here, we identify two genes required for the major excitatory current found at the C. elegans NMJ: acr-16, which encodes a nicotinic AChR subunit homologous to the vertebrate alpha7 subunit, and cam-1, which encodes a Ror receptor tyrosine kinase. acr-16 mutants lack fast cholinergic current at the NMJ and exhibit synthetic behavioral deficits with other known AChR mutants. In cam-1 mutants, ACR-16 is mislocalized and ACR-16-dependent currents are disrupted. The postsynaptic deficit in cam-1 mutants is accompanied by alterations in the distribution of cholinergic vesicles and associated synaptic proteins. We hypothesize that CAM-1 contributes to the localization or stabilization of postsynaptic ACR-16 receptors and presynaptic release sites.

ENU mutagenesis identifies mice with cardiac fibrosis and hepatic steatosis caused by a mutation in the mitochondrial trifunctional protein beta-subunit.

PubMed

Kao, Hsiao-Jung; Cheng, Ching-Feng; Chen, Yen-Hui; Hung, Shuen-Iu; Huang, Cheng-Chih; Millington, David; Kikuchi, Tateki; Wu, Jer-Yuarn; Chen, Yuan-Tsong

2006-12-15

Using the metabolomics-guided screening coupled to N-ethyl-N-nitrosourea-mediated mutagenesis, we identified mice that exhibited elevated levels of long-chain acylcarnitines. Whole genome homozygosity mapping with 262 SNP markers mapped the disease gene to chromosome 5 where candidate genes Hadha and Hadhb, encoding the mitochondria trifunctional protein (MTP) alpha- and beta-subunits, respectively, are located. Direct sequencing revealed a normal alpha-subunit, but detected a nucleotide T-to-A transversion in exon 14 (c.1210T>A) of beta-subunit (Hadhb) which resulted in a missense mutation of methionine to lysine (M404K). Western blot analysis showed a significant reduction of both the alpha- and beta-subunits, consistent with reduced enzyme activity in both the long-chain 3-hydroxyacyl-CoA dehydrogenase and the long-chain 3-ketoacyl-CoA thiolase activities. These mice had a decreased weight gain and cardiac arrhythmias which manifested from a prolonged PR interval to a complete atrio-ventricular dissociation, and died suddenly between 9 and 16 months of age. Histopathological studies showed multifocal cardiac fibrosis and hepatic steatosis. This mouse model will be useful to further investigate the mechanisms underlying arrhythmogenesis relating to lipotoxic cardiomyopathy and to investigate pathophysiology and treatment strategies for human MTP deficiency.

Small subunits of RNA polymerase: localization, levels and implications for core enzyme composition.

PubMed

Doherty, Geoff P; Fogg, Mark J; Wilkinson, Anthony J; Lewis, Peter J

2010-12-01

Bacterial RNA polymerases (RNAPs) contain several small auxiliary subunits known to co-purify with the core α, β and β' subunits. The ω subunit is conserved between Gram-positive and Gram-negative bacteria, while the δ subunit is conserved within, but restricted to, Gram-positive bacteria. Although various functions have been assigned to these subunits via in vitro assays, very little is known about their in vivo roles. In this work we constructed a pair of vectors to investigate the subcellular localization of the δ and ω subunits in Bacillus subtilis with respect to the core RNAP. We found these subunits to be closely associated with RNAP involved in transcribing both mRNA and rRNA operons. Quantification of these subunits revealed δ to be present at equimolar levels with RNAP and ω to be present at around half the level of core RNAP. For comparison, the localization and quantification of RNAP β' and ω subunits in Escherichia coli was also investigated. Similar to B. subtilis, β' and ω closely associated with the nucleoid and formed subnucleoid regions of high green fluorescent protein intensity, but, unlike ω in B. subtilis, ω levels in E. coli were close to parity with those of β'. These results indicate that δ is likely to be an integral RNAP subunit in Gram-positives, whereas ω levels differ substantially between Gram-positives and -negatives. The ω subunit may be required for RNAP assembly and subsequently be turned over at different rates or it may play roles in Gram-negative bacteria that are performed by other factors in Gram-positives.

Cognitive Function in Prepubertal Children with Obstructive Sleep Apnea: A Modifying Role for NADPH Oxidase p22 Subunit Gene Polymorphisms?

PubMed Central

Khalyfa, Abdelnaby; Capdevila, Oscar Sans; Kheirandish-Gozal, Leila; Khalyfa, Ahamed A.; Kim, Jinkwan

2012-01-01

Abstract Pediatric obstructive sleep apnea (OSA) may lead to neurocognitive dysfunction, but not in everyone affected. The frequencies of NADPH oxidase (NOX) polymorphisms in the p22phox subunit were similar between children with OSA and controls, except for rs6520785 and rs4673, the latter being significantly more frequent among the OSA children without deficits than with deficits (p<0.02). Similarly, 8-hydroxydeoxyguanine urine levels and NOX activity were lower among children without cognitive deficits and particularly among those with the rs4673 polymorphism. Thus, polymorphisms within the NOX gene or its functional subunits may account for important components of the variance in cognitive function deficits associated with OSA in children. Antioxid. Redox Signal. 16, 171–177. PMID:21902598

Epitopes from two soybean glycinin subunits antigenic in pigs

USDA-ARS?s Scientific Manuscript database

Background: Glycinin is a seed storage protein in soybean (Glycine max) that is allergenic in pigs. Glycinin is a hexamer composed of subunits consisting of a basic and acidic portion joined by disulfide bridges. There are 5 glycinin subunits designated Gy1-Gy5. Results: Twenty seven out of 30 pi...

Prevalence and relevance of the positivity of skin prick testing in patients with chronic urticaria.

PubMed

Kulthanan, Kanokvalai; Jiamton, Sukhum; Rutnin, Ni-on; Insawang, Metavee; Pinkaew, Sumrauy

2008-06-01

Many patients with chronic urticaria (CU) worry that foods or other allergens are responsible for their urticaria. Skin prick testing (SPT) may be one of the investigations used to provide a clear illustration. The purpose of our study was to assess the prevalence of positivity of SPT to food allergens and aeroallergens and their relevance in patients with CU, in order to demonstrate the diagnostic value of SPT in CU. We retrospectively reviewed case record forms of patients with chronic ordinary urticaria who underwent SPT in the Urticaria Clinic, Siriraj Hospital, during the period 2000-2004. The studied allergens included 16 food allergens and 12 aeroallergens. Eighty-eight patients were enrolled. The prevalence of positive SPT among patients with CU was 47.7%. Patients who had personal histories of atopy had statistically significant positive SPT results compared with patients who had negative SPT. Of 88 patients, 26 patients (30%) gave positive results to food allergens, 36 patients (41%) gave positive results to aeroallergens and 20 patients (22.7%) gave positive results to both food and aeroallergens. One-third of the subjects (34.6%) who had positive SPT results to food allergens had clinical relevance of food allergy in some systems but only one patient had clinical relevance of food-induced urticaria. Half of the patients who had a history of aeroallergen sensitivity gave a positive SPT response for aeroallergens; however, there was no clinical relevance to their CU. Our study showed that the prevalence of positive SPT to food allergens and aeroallergens in patients with CU was common but had little clinical relevance to CU.

Fact Sheet: Final Air Toxics Standards for Area Sources in the Chemical Manufacturing Industry

EPA Pesticide Factsheets

Fact sheet on the national air toxics standards issued October 16, 2009 by the Environmental Protection Agency (EPA) for smaller-emitting sources, known as area sources, in the chemical manufacturing industry.

Gal4-VP16 directs ATP-independent chromatin reorganization in a yeast chromatin assembly system.

PubMed

Robinson, Karen M; Schultz, Michael C

2005-03-22

Major insights into the regulation of chromatin organization have stemmed from biochemical studies using Gal4-VP16, a chimeric transcriptional activator in which the DNA binding domain of Gal4p is fused to the activation domain of viral protein VP16. Unexpectedly, given previous intensive efforts to understand how Gal4-VP16 functions in the context of chromatin, we have uncovered a new mode of chromatin reorganization that is dependent on Gal4-VP16. This reorganization is performed by an activity in a crude DEAE (CD) fraction from budding yeast which also supports ATP-dependent assembly of physiologically spaced nucleosome arrays. Biochemical analysis reveals that the activity tightly associates with chromatin and reorganizes nucleosome arrays by a mechanism which is insensitive to ATP depletion after nucleosome assembly. It generates a chromatin organization in which a nucleosome is stably positioned immediately adjacent to Gal4p binding sites in the template DNA. Individual deletion of genes previously implicated in chromatin assembly and remodeling, namely, the histone chaperones NAP1, ASF1, and CAC1 and the SNF2-like DEAD/H ATPases SNF2, ISW1, ISW2, CHD1, SWR1, YFR038w, and SPT20, does not significantly perturb reorganization. Therefore, Gal4-VP16-directed chromatin reorganization in yeast can occur by an ATP-independent mechanism that does not require SAGA, SWI/SNF, Isw1, or Isw2 chromatin remodeling complexes.

Disordered organic electronic materials based on non-benzenoid 1,6-methano[10]annulene rings

DOEpatents

Tovar, John D; Streifel, Benjamin C; Peart, Patricia A

2014-10-07

Conjugated polymers and small molecules including the nonplanar aromatic 1,6-methano[10]annulene ring structure along with aromatic subunits, such as diketopyrrolopyrrole, and 2,1,3-benzothiadiazole, substituted with alkyl chains in a "Tail In," "Tail Out," or "No Tail" regiochemistry are disclosed.

Inactivation properties of voltage-gated K+ channels altered by presence of beta-subunit.

PubMed

Rettig, J; Heinemann, S H; Wunder, F; Lorra, C; Parcej, D N; Dolly, J O; Pongs, O

1994-05-26

Structural and functional diversity of voltage-gated Kv1-type potassium channels in rat brain is enhanced by the association of two different types of subunits, the membrane-bound, poreforming alpha-subunits and a peripheral beta-subunit. We have cloned a beta-subunit (Kv beta 1) that is specifically expressed in the rat nervous system. Association of Kv beta 1 with alpha-subunits confers rapid A-type inactivation on non-inactivating Kv1 channels (delayed rectifiers) in expression systems in vitro. This effect is mediated by an inactivating ball domain in the Kv beta 1 amino terminus.

Diet myths and facts

MedlinePlus

Obesity - diet myths and facts; Overweight - diet myths and fact; Weight-loss diet myths and facts ... evidence: using the proposed effect of breakfast on obesity to show 2 practices that distort scientific evidence. ...

The mongoose acetylcholine receptor alpha-subunit: analysis of glycosylation and alpha-bungarotoxin binding.

PubMed

Asher, O; Jensen, B S; Lupu-Meiri, M; Oron, Y; Fuchs, S

1998-04-17

The mongoose AChR alpha-subunit has been cloned and shown to be highly homologous to other AChR alpha-subunits, with only six differences in amino acid residues at positions that are conserved in animal species that bind alpha-bungarotoxin (alpha-BTX). Four of these six substitutions cluster in the ligand binding site, and one of them, Asn-187, forms a consensus N-glycosylation site. The mongoose glycosylated alpha-subunit has a higher apparent molecular mass than that of the rat glycosylated alpha-subunit, probably resulting from the additional glycosylation at Asn-187 of the mongoose subunit. The in vitro translated mongoose alpha-subunit, in a glycosylated or non-glycosylated form, does not bind alpha-BTX, indicating that lack of alpha-BTX binding can be achieved also in the absence of glycosylation.

Proteopedia Entry: The Large Ribosomal Subunit of "Haloarcula Marismortui"

ERIC Educational Resources Information Center

Decatur, Wayne A.

2010-01-01

This article presents a "Proteopedia" page that shows the refined version of the structure of the "Haloarcula" large ribosomal subunit as solved by the laboratories of Thomas Steitz and Peter Moore. The landmark structure is of great impact as it is the first atomic-resolution structure of the highly conserved ribosomal subunit which harbors…

How glutamate receptor subunits mix and match: details uncovered.

PubMed

Hansen, Kasper B; Traynelis, Stephen F

2011-07-28

Until now, the atomic details explaining why certain subunits prefer to coassemble has been lacking in our understanding of glutamate receptor biogenesis. In this issue, Kumar et al. describe the structural basis by which preferential subunit assembly occurs for homomeric and heteromeric kainate-type glutamate receptors. Copyright © 2011 Elsevier Inc. All rights reserved.

Pea chloroplast DNA encodes homologues of Escherichia coli ribosomal subunit S2 and the beta'-subunit of RNA polymerase.

PubMed Central

Cozens, A L; Walker, J E

1986-01-01

The nucleotide sequence has been determined of a segment of 4680 bases of the pea chloroplast genome. It adjoins a sequence described elsewhere that encodes subunits of the F0 membrane domain of the ATP-synthase complex. The sequence contains a potential gene encoding a protein which is strongly related to the S2 polypeptide of Escherichia coli ribosomes. It also encodes an incomplete protein which contains segments that are homologous to the beta'-subunit of E. coli RNA polymerase and to yeast RNA polymerases II and III. PMID:3530249

Heterosubunit composition and crystal structures of a novel bacterial M16B metallopeptidase.

PubMed

Maruyama, Yukie; Chuma, Asako; Mikami, Bunzo; Hashimoto, Wataru; Murata, Kousaku

2011-03-18

Three subfamilies of metallopeptidase family M16 enzymes--M16A, M16B, and M16C--are widely distributed among eukaryotes and prokaryotes. SPH2681, a periplasmic M16B protein found in Sphingomonas sp. strain A1, contains an HXXEH motif essential for Zn(2+) binding and catalytic activity. SPH2682 is another member of M16B, which lacks the metal-binding motif but conserves an active-site R/Y pair commonly found in the C-terminal half of M16 enzymes. Two genes coding for SPH2681 and SPH2682 assemble into a single operon in the bacterial genome. This study determined SPH2681 to be constitutively expressed in strain A1 cells grown on different carbon sources, suggesting a more general cellular function. SPH2681 and SPH2681/SPH2682 were overexpressed in Escherichia coli, purified, and characterized. SPH2681 was found to associate with SPH2682, forming a heterosubunit enzyme with peptidase activity, while SPH2681 alone exhibited no enzymatic activity. X-ray crystallography of the SPH2681/SPH2682 complex revealed two conformations (open and closed heterodimeric forms) within the same crystal. Compared with the closed form, the open form contains two subunits rotated away from each other by approximately 8°, increasing the distance between the zinc ion and active-site residues by up to 8 Å. In addition, many hydrogen bonds are formed or broken on change between the conformations of the heterodimers, suggesting that subunit dynamics is a prerequisite for catalysis. To our knowledge, this is the first report on both conformational forms of the same M16 peptidase, providing a unique insight into the general proteolytic mechanism of M16 proteases. Copyright © 2011 Elsevier Ltd. All rights reserved.

The Role of Auxiliary Subunits for the Functional Diversity of Voltage-Gated Calcium Channels

PubMed Central

Campiglio, Marta; Flucher, Bernhard E

2015-01-01

Voltage-gated calcium channels (VGCCs) represent the sole mechanism to convert membrane depolarization into cellular functions like secretion, contraction, or gene regulation. VGCCs consist of a pore-forming α1 subunit and several auxiliary channel subunits. These subunits come in multiple isoforms and splice-variants giving rise to a stunning molecular diversity of possible subunit combinations. It is generally believed that specific auxiliary subunits differentially regulate the channels and thereby contribute to the great functional diversity of VGCCs. If auxiliary subunits can associate and dissociate from pre-existing channel complexes, this would allow dynamic regulation of channel properties. However, most auxiliary subunits modulate current properties very similarly, and proof that any cellular calcium channel function is indeed modulated by the physiological exchange of auxiliary subunits is still lacking. In this review we summarize available information supporting a differential modulation of calcium channel functions by exchange of auxiliary subunits, as well as experimental evidence in support of alternative functions of the auxiliary subunits. At the heart of the discussion is the concept that, in their native environment, VGCCs function in the context of macromolecular signaling complexes and that the auxiliary subunits help to orchestrate the diverse protein–protein interactions found in these calcium channel signalosomes. Thus, in addition to a putative differential modulation of current properties, differential subcellular targeting properties and differential protein–protein interactions of the auxiliary subunits may explain the need for their vast molecular diversity. J. Cell. Physiol. 999: 00–00, 2015. © 2015 The Authors. Journal of Cellular Physiology Published by Wiley Periodicals, Inc. J. Cell. Physiol. 230: 2019–2031, 2015. © 2015 Wiley Periodicals, Inc. PMID:25820299

The depletion of F1 subunit ε in yeast leads to an uncoupled respiratory phenotype that is rescued by mutations in the proton-translocating subunits of F0

PubMed Central

Tetaud, Emmanuel; Godard, François; Giraud, Marie-France; Ackerman, Sharon H.; di Rago, Jean-Paul

2014-01-01

The central stalk of the ATP synthase is an elongated hetero-oligomeric structure providing a physical connection between the catalytic sites in F1 and the proton translocation channel in F0 for energy transduction between the two subdomains. The shape of the central stalk and relevance to energy coupling are essentially the same in ATP synthases from all forms of life, yet the protein composition of this domain changed during evolution of the mitochondrial enzyme from a two- to a three-subunit structure (γ, δ, ε). Whereas the mitochondrial γ- and δ-subunits are homologues of the bacterial central stalk proteins, the deliberate addition of subunit ε is poorly understood. Here we report that down-regulation of the gene (ATP15) encoding the ε-subunit rapidly leads to lethal F0-mediated proton leaks through the membrane because of the loss of stability of the ATP synthase. The ε-subunit is thus essential for oxidative phosphorylation. Moreover, mutations in F0 subunits a and c, which slow the proton translocation rate, are identified that prevent ε-deficient ATP synthases from dissipating the electrochemical potential. Cumulatively our data lead us to propose that the ε-subunit evolved to permit operation of the central stalk under the torque imposed at the normal speed of proton movement through mitochondrial F0. PMID:24451261

Influvac, a trivalent inactivated subunit influenza vaccine.

PubMed

Zuccotti, Gian Vincenzo; Fabiano, Valentina

2011-01-01

Influenza represents a major sanitary and socio-economic burden and vaccination is universally considered the most effective strategy for preventing the disease and its complications. Traditional influenza vaccines have been on the market since the late 1940s, with million of doses administered annually worldwide, and demonstrated a substantial efficacy and safety. The trivalent inactivated subunit vaccine has been available for more than 25 years and has been studied in healthy children, adults and the elderly and in people affected by underlying chronic medical conditions. We describe vaccine technology focusing on subunit vaccine production procedures and mode of action and provide updated information on efficacy and safety available data. A review of efficacy and safety data in healthy subjects and in high risk populations from major sponsor- and investigator-driven studies. The vaccine showed a good immunogenicity and a favorable safety profile in all target groups. In the panorama of actually available influenza vaccines, trivalent inactivated subunit vaccine represents a well-established tool for preventing flu and the associated complications.

Nicotinic acetylcholine receptor alpha5 subunits modulate oxotremorine-induced salivation and tremor.

PubMed

Wang, Ningshan; Orr-Urtreger, Avi; Chapman, Joab; Rabinowitz, Ruth; Korczyn, Amos D

2004-07-15

Neuronal nicotinic acetylcholine receptors (nAChRs) are composed of 12 subunits (alpha2-alpha10 and beta2-beta4). alpha5 Subunits, expressed throughout the central nervous system (CNS) and the autonomic nervous system (ANS), possess unique pharmacological properties. The effects of oxotremorine (OXO) on autonomic functions and tremor were examined in mice lacking alpha5 nAChR subunits (alpha5-/-) and compared with those in wild-type (WT) control mice. The alpha5-/- mice showed significantly increased salivation and tremor responses to OXO. The hypothermia, bradycardia and defecation induced by OXO were of similar magnitudes in the two mouse strains. The enhanced OXO effects in alpha5-/- mice indicate inhibitory effects of alpha5 subunits in autonomic ganglia, and support the participation of these subunits in cholinergic transmission in autonomic ganglia.

First Weak-lensing Results from “See Change”: Quantifying Dark Matter in the Two z ≳ 1.5 High-redshift Galaxy Clusters SPT-CL J2040-4451 and IDCS J1426+3508

NASA Astrophysics Data System (ADS)

Jee, M. James; Ko, Jongwan; Perlmutter, Saul; Gonzalez, Anthony; Brodwin, Mark; Linder, Eric; Eisenhardt, Peter

2017-10-01

We present a weak-lensing study of SPT-CL J2040-4451 and IDCS J1426+3508 at z = 1.48 and 1.75, respectively. The two clusters were observed in our “See Change” program, a Hubble Space Telescope survey of 12 massive high-redshift clusters aimed at high-z supernova measurements and weak-lensing estimation of accurate cluster masses. We detect weak but significant galaxy shape distortions using infrared images from the Wide Field Camera 3 (WFC3), which has not yet been used for weak-lensing studies. Both clusters appear to possess relaxed morphology in projected mass distribution, and their mass centroids agree nicely with those defined by both the galaxy luminosity and X-ray emission. Using a Navarro-Frenk-White profile, for which we assume that the mass is tightly correlated with the concentration parameter, we determine the masses of SPT-CL J2040-4451 and IDCS J1426 + 3508 to be {M}200={8.6}-1.4+1.7× {10}14 {M}⊙ and {2.2}-0.7+1.1× {10}14 {M}⊙ , respectively. The weak-lensing mass of SPT-CL J2040-4451 shows that the cluster is clearly a rare object. Adopting the central value, the expected abundance of such a massive cluster at z≳ 1.48 is only ˜ 0.07 in the parent 2500 sq. deg. survey. However, it is yet premature to claim that the presence of this cluster creates a serious tension with the current ΛCDM paradigm unless that tension will remain in future studies after marginalizing over many sources of uncertainties such as the accuracy of the mass function and the mass-concentration relation at the high-mass end. The mass of IDCS J1426+3508 is in excellent agreement with our previous Advanced Camera for Surveys-based weak-lensing result, while the much higher source density from our WFC3 imaging data makes the current statistical uncertainty ˜ 40% smaller.

Cloning and characterization of two novel zebrafish P2X receptor subunits.

PubMed

Diaz-Hernandez, Miguel; Cox, Jane A; Migita, Keisuke; Haines, William; Egan, Terrance M; Voigt, Mark M

2002-07-26

In this report we describe the cloning and characterization of two P2X receptor subunits cloned from the zebrafish (Danio rerio). Primary sequence analysis suggests that one cDNA encodes an ortholog of the mammalian P2X(4) subunit and the second cDNA encodes the ortholog of the mammalian P2X(5) subunit. The zP2X(4) subunit forms a homo-oligomeric receptor that displays a low affinity for ATP (EC(50)=274+/-48 microM) and very low affinity (EC(50)>500 microM) for other purinergic ligands such as alphabetameATP, suramin, and PPADS. As seen with the mammalian orthologs, the zP2X(5) subunit forms a homo-oligomeric receptor that yields very small whole-cell currents (<20pA), making determination of an EC(50) problematic. Both subunit genes were physically mapped onto the zebrafish genome using radiation hybrid analysis of the T51 panel, with the zp2x4 localized to LG21 and zp2x5 to LG5.

Quantification of Transthyretin Kinetic Stability in Human Plasma Using Subunit Exchange

PubMed Central

2015-01-01

The transthyretin (TTR) amyloidoses are a group of degenerative diseases caused by TTR aggregation, requiring rate-limiting tetramer dissociation. Kinetic stabilization of TTR, by preferential binding of a drug to the native tetramer over the dissociative transition state, dramatically slows the progression of familial amyloid polyneuropathy. An established method for quantifying the kinetic stability of recombinant TTR tetramers in buffer is subunit exchange, in which tagged TTR homotetramers are added to untagged homotetramers at equal concentrations to measure the rate at which the subunits exchange. Herein, we report a subunit exchange method for quantifying the kinetic stability of endogenous TTR in human plasma. The subunit exchange reaction is initiated by the addition of a substoichiometric quantity of FLAG-tagged TTR homotetramers to endogenous TTR in plasma. Aliquots of the subunit exchange reaction, taken as a function of time, are then added to an excess of a fluorogenic small molecule, which immediately arrests further subunit exchange. After binding, the small molecule reacts with the TTR tetramers, rendering them fluorescent and detectable in human plasma after subsequent ion exchange chromatography. The ability to report on the extent of TTR kinetic stabilization resulting from treatment with oral tafamidis is important, especially for selection of the appropriate dose for patients carrying rare mutations. This method could also serve as a surrogate biomarker for the prediction of the clinical outcome. Subunit exchange was used to quantify the stabilization of WT TTR from senile systemic amyloidosis patients currently being treated with tafamidis (20 mg orally, once daily). TTR kinetic stability correlated with the tafamidis plasma concentration. PMID:24661308

The eukaryotic RNA exosome: same scaffold but variable catalytic subunits.

PubMed

Lykke-Andersen, Søren; Tomecki, Rafal; Jensen, Torben Heick; Dziembowski, Andrzej

2011-01-01

The RNA exosome is a versatile ribonucleolytic protein complex that participates in a multitude of cellular RNA processing and degradation events. It consists of an invariable nine-subunit core that associates with a variety of enzymatically active subunits and co-factors. These contribute to or even provide the catalytic activity and substrate specificity of the complex. The S. cerevisiae exosome has been intensively studied since its discovery in 1997 and thus serves as the archetype of eukaryotic exosomes. Notably, its catalytic potential, derived exclusively from associated subunits, differs between the nuclear and cytoplasmic versions of the complex. The same holds true for other eukaryotes, however, recent discoveries from various laboratories including our own have revealed that there are variations on this theme. Here, we review the latest findings concerning catalytic subunits of eukaryotic exosomes, and we discuss the apparent need for differential composition and subcellular distribution of exosome variants.

An Approach to Flooding Inundation Combining the Streamflow Prediction Tool (SPT) and Downscaled Soil Moisture

NASA Astrophysics Data System (ADS)

Cotterman, K. A.; Follum, M. L.; Pradhan, N. R.; Niemann, J. D.

2017-12-01

Flooding impacts numerous aspects of society, from localized flash floods to continental-scale flood events. Many numerical flood models focus solely on riverine flooding, with some capable of capturing both localized and continental-scale flood events. However, these models neglect flooding away from channels that are related to excessive ponding, typically found in areas with flat terrain and poorly draining soils. In order to obtain a holistic view of flooding, we combine flood results from the Streamflow Prediction Tool (SPT), a riverine flood model, with soil moisture downscaling techniques to determine if a better representation of flooding is obtained. This allows for a more holistic understanding of potential flood prone areas, increasing the opportunity for more accurate warnings and evacuations during flooding conditions. Thirty-five years of near-global historical streamflow is reconstructed with continental-scale flow routing of runoff from global land surface models. Elevation data was also obtained worldwide, to establish a relationship between topographic attributes and soil moisture patterns. Derived soil moisture data is validated against observed soil moisture, increasing confidence in the ability to accurately capture soil moisture patterns. Potential flooding situations can be examined worldwide, with this study focusing on the United States, Central America, and the Philippines.

Arginine 26 and aspartic acid 69 of the regulatory subunit are key residues of subunits interaction of acetohydroxyacid synthase isozyme III from E. coli.

PubMed

Zhao, Yuefang; Wen, Xin; Niu, Congwei; Xi, Zhen

2012-11-05

Acetohydroxyacid synthase (AHAS), which catalyzes the first step in the biosynthesis of branched-chain amino acids, is composed of catalytic and regulatory subunits. The enzyme exhibits full activity only when the regulatory subunit (RSU) binds to the catalytic subunit (CSU). However, the crystal structure of the holoenzyme has not been reported yet, and the molecular interaction between the CSU and RSU is also unknown. Herein, we introduced a global-surface, site-directed labeling scanning method to determine the potential interaction region of the RSU. This approach relies on the insertion of a bulky fluorescent probe at the designated site on the surface of the RSU to cause a dramatic change in holoenzyme activity by perturbing subunit interaction. Then, the key amino acid residues in the potential interaction regions were identified by site-directed mutagenesis. Compared to the wild-type, the single-point mutants R26A and D69A showed 54 and 64 % activity, respectively, whereas the double mutant (R26A+D69A) gave 14 %, thus suggesting that residues Arg26 and Asp69 are the key residues of subunit interaction with cooperative action. Additionally, the results of GST pull-down assays and pH-dependence experiments suggested that polar interaction is the main force for subunits interaction. A plausible protein-protein interaction model of the holoenzyme of Escherichia coli AHAS III is proposed, based on the mutagenesis and protein docking studies. The protocol established here should be useful for the identification of the molecular interactions between proteins. Copyright © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Effects of detergents on ribosomal precursor subunits of Bacillus megaterium.

PubMed

Body, A; Brownstein, B H

1978-01-01

Cell extracts prepared by osmotic lysis of protoplasts were analyzed by sucrose gradient sedimentation. In the absence of detergents, ribosomal precursor particles were found in a gradient fraction which sedimented faster than mature 50S subunits and in two other fractions coincident with mature 50S and 30S ribosomal subunits. Phospholipid, an indicator of membrane, was shown to be associated with only the fastest-sedimenting ribosomal precursor particle fraction. After the extracts were treated with detergents, all phospholipid was found at the top of the gradients. Brij 58, Triton X-100, and Nonidet P-40 did not cause a change in the sedimentation values of precursors; however, the detergents deoxycholate or LOC (Amway Corp.) disrupted the fastest-sedimenting precursor and converted the ribosomal precursor subunits which sedimented at the 50S and 30S positions to five different classes of more slowly sedimenting particles. Earlier reports on the in vivo assembly of ribosomal subunits have shown that several stages of ribosomal precursor subunits exist, and, in the presence of the detergents deoxycholate and LOC, which had been used to prepare cell extracts, the precursors sedimented more slowly. Our data are consistent with the hypothesis that those detergents selectively modify the structure of ribosomal precursors and lend further support to the hypothesis that the in vivo ribosomal precursor subunits have 50S and 30S sedimentation values. In addition, these data support the idea that the ribosomal precursor particles found in the fast-sedimenting fraction may constitute a unique precursor fraction.

Effects of Detergents on Ribosomal Precursor Subunits of Bacillus megaterium

PubMed Central

Body, Barbara A.; Brownstein, Bernard H.

1978-01-01

Cell extracts prepared by osmotic lysis of protoplasts were analyzed by sucrose gradient sedimentation. In the absence of detergents, ribosomal precursor particles were found in a gradient fraction which sedimented faster than mature 50S subunits and in two other fractions coincident with mature 50S and 30S ribosomal subunits. Phospholipid, an indicator of membrane, was shown to be associated with only the fastest-sedimenting ribosomal precursor particle fraction. After the extracts were treated with detergents, all phospholipid was found at the top of the gradients. Brij 58, Triton X-100, and Nonidet P-40 did not cause a change in the sedimentation values of precursors; however, the detergents deoxycholate or LOC (Amway Corp.) disrupted the fastest-sedimenting precursor and converted the ribosomal precursor subunits which sedimented at the 50S and 30S positions to five different classes of more slowly sedimenting particles. Earlier reports on the in vivo assembly of ribosomal subunits have shown that several stages of ribosomal precursor subunits exist, and, in the presence of the detergents deoxycholate and LOC, which had been used to prepare cell extracts, the precursors sedimented more slowly. Our data are consistent with the hypothesis that those detergents selectively modify the structure of ribosomal precursors and lend further support to the hypothesis that the in vivo ribosomal precursor subunits have 50S and 30S sedimentation values. In addition, these data support the idea that the ribosomal precursor particles found in the fast-sedimenting fraction may constitute a unique precursor fraction. PMID:412833

CRISPR Genome-Wide Screening Identifies Dependence on the Proteasome Subunit PSMC6 for Bortezomib Sensitivity in Multiple Myeloma.

PubMed

Shi, Chang-Xin; Kortüm, K Martin; Zhu, Yuan Xiao; Bruins, Laura A; Jedlowski, Patrick; Votruba, Patrick G; Luo, Moulun; Stewart, Robert A; Ahmann, Jonathan; Braggio, Esteban; Stewart, A Keith

2017-12-01

Bortezomib is highly effective in the treatment of multiple myeloma; however, emergent drug resistance is common. Consequently, we employed CRISPR targeting 19,052 human genes to identify unbiased targets that contribute to bortezomib resistance. Specifically, we engineered an RPMI8226 multiple myeloma cell line to express Cas9 infected by lentiviral vector CRISPR library and cultured derived cells in doses of bortezomib lethal to parental cells. Sequencing was performed on surviving cells to identify inactivated genes responsible for drug resistance. From two independent whole-genome screens, we selected 31 candidate genes and constructed a second CRISPR sgRNA library, specifically targeting each of these 31 genes with four sgRNAs. After secondary screening for bortezomib resistance, the top 20 "resistance" genes were selected for individual validation. Of these 20 targets, the proteasome regulatory subunit PSMC6 was the only gene validated to reproducibly confer bortezomib resistance. We confirmed that inhibition of chymotrypsin-like proteasome activity by bortezomib was significantly reduced in cells lacking PSMC6. We individually investigated other members of the PSMC group (PSMC1 to 5) and found that deficiency in each of those subunits also imparts bortezomib resistance. We found 36 mutations in 19S proteasome subunits out of 895 patients in the IA10 release of the CoMMpass study (https://themmrf.org). Our findings demonstrate that the PSMC6 subunit is the most prominent target required for bortezomib sensitivity in multiple myeloma cells and should be examined in drug-refractory populations. Mol Cancer Ther; 16(12); 2862-70. ©2017 AACR . ©2017 American Association for Cancer Research.

16 CFR 2.32 - Agreement.

Code of Federal Regulations, 2013 CFR

2013-01-01

... 16 Commercial Practices 1 2013-01-01 2013-01-01 false Agreement. 2.32 Section 2.32 Commercial... Consent Order Procedure § 2.32 Agreement. Every agreement in settlement of a Commission complaint shall... of findings of fact and conclusions of law. Every agreement also shall waive further procedural steps...

16 CFR 2.32 - Agreement.

Code of Federal Regulations, 2010 CFR

2010-01-01

... 16 Commercial Practices 1 2010-01-01 2010-01-01 false Agreement. 2.32 Section 2.32 Commercial... Consent Order Procedure § 2.32 Agreement. Every agreement in settlement of a Commission complaint shall... of findings of fact and conclusions of law. Every agreement also shall waive further procedural steps...

16 CFR 2.32 - Agreement.

Code of Federal Regulations, 2012 CFR

2012-01-01

... 16 Commercial Practices 1 2012-01-01 2012-01-01 false Agreement. 2.32 Section 2.32 Commercial... Consent Order Procedure § 2.32 Agreement. Every agreement in settlement of a Commission complaint shall... of findings of fact and conclusions of law. Every agreement also shall waive further procedural steps...

16 CFR 2.32 - Agreement.

Code of Federal Regulations, 2011 CFR

2011-01-01

... 16 Commercial Practices 1 2011-01-01 2011-01-01 false Agreement. 2.32 Section 2.32 Commercial... Consent Order Procedure § 2.32 Agreement. Every agreement in settlement of a Commission complaint shall... of findings of fact and conclusions of law. Every agreement also shall waive further procedural steps...

16 CFR 2.32 - Agreement.

Code of Federal Regulations, 2014 CFR

2014-01-01

... 16 Commercial Practices 1 2014-01-01 2014-01-01 false Agreement. 2.32 Section 2.32 Commercial... Consent Order Procedure § 2.32 Agreement. Every agreement in settlement of a Commission complaint shall... of findings of fact and conclusions of law. Every agreement also shall waive further procedural steps...

Rat lung glutathione S-transferases. Evidence for two distinct types of 22000-Mr subunits.

PubMed Central

Singh, S V; Partridge, C A; Awasthi, Y C

1984-01-01

Two immunologically distinct types of 22000-Mr subunits are present in rat lung glutathione S-transferases. One of these subunits is probably similar to Ya subunits of rat liver glutathione S-transferases, whereas the other subunit Ya' is immunologically distinct. Glutathione S-transferase II (pI7.2) of rat lung is a heterodimer (YaYa') of these subunits, and glutathione S-transferase VI (pI4.8) of rat lung is a homodimer of Ya' subunits. On hybridization in vitro of the subunits of glutathione S-transferase II of rat lung three active dimers having pI values 9.4, 7.2 and 4.8 are obtained. Immunological properties and substrate specificities indicate that the hybridized enzymes having pI7.2 and 4.8 correspond to glutathione S-transferases II and VI of rat lung respectively. Images Fig. 1. Fig. 5. PMID:6433888

Exact Length Distribution of Filamentous Structures Assembled from a Finite Pool of Subunits.

PubMed

Harbage, David; Kondev, Jané

2016-07-07

Self-assembling filamentous structures made of protein subunits are ubiquitous in cell biology. These structures are often highly dynamic, with subunits in a continuous state of flux, binding to and falling off of filaments. In spite of this constant turnover of their molecular parts, many cellular structures seem to maintain a well-defined size over time, which is often required for their proper functioning. One widely discussed mechanism of size regulation involves the cell maintaining a finite pool of protein subunits available for assembly. This finite pool mechanism can control the length of a single filament by having assembly proceed until the pool of free subunits is depleted to the point when assembly and disassembly are balanced. Still, this leaves open the question of whether the same mechanism can provide size control for multiple filamentous structures that are assembled from a common pool of protein subunits, as is often the case in cells. We address this question by solving the steady-state master equation governing the stochastic assembly and disassembly of multifilament structures made from a shared finite pool of subunits. We find that, while the total number of subunits within a multifilament structure is well-defined, individual filaments within the structure have a wide, power-law distribution of lengths. We also compute the phase diagram for two multifilament structures competing for the same pool of subunits and identify conditions for coexistence when both have a well-defined size. These predictions can be tested in cell experiments in which the size of the subunit pool or the number of filament nucleators is tuned.

Non-surgical and supportive periodontal therapy: predictors of compliance

PubMed Central

Delatola, Chrysoula; Adonogianaki, Evagelia; Ioannidou, Effie

2015-01-01

Aim To identify predictors of compliance during non-surgical and supportive periodontal therapy (SPT). Materials and Methods In this retrospective study, demographic, dental, medical data of 427 new patients in a private practice were collected. Data were analysed in statistical models with non-surgical therapy and SPT compliance used as dependent variables. Results Of the 427 patients, 17.3% never agreed to initial therapy, 10.7% never completed therapy and 20.8% completed treatment, but never entered SPT. Of the 218 SPT patients, 56% became non-attenders after a period of 20 months, 33% were erratic attenders and 10.5% were regular attenders until the end of the observation period (5.5–6.5 years). Patients became erratic attenders after a mean period of regular attendance of 18.1 ± 16.2 months, whereas 49.6% of the patients, who abandoned SPT, were regular attenders until the time they stopped. In a univariate correlation model, periodontal disease severity emerged as a significant predictor of the completion of non-surgical periodontal therapy (p = 0.01). In a multivariate linear regression model, smoking was negatively associated with SPT compliance (p = 0.047). Conclusions A low compliance of the population was observed. Smoking and periodontal disease severity represented significant, but modest modifiers of a patient compliance with SPT and initial therapy respectively. PMID:24813661

Factors Affecting Nuclear Export of the 60S Ribosomal Subunit In Vivo

PubMed Central

Stage-Zimmermann, Tracy; Schmidt, Ute; Silver, Pamela A.

2000-01-01

In Saccharomyces cerevisiae, the 60S ribosomal subunit assembles in the nucleolus and then is exported to the cytoplasm, where it joins the 40S subunit for translation. Export of the 60S subunit from the nucleus is known to be an energy-dependent and factor-mediated process, but very little is known about the specifics of its transport. To begin to address this problem, an assay was developed to follow the localization of the 60S ribosomal subunit in S. cerevisiae. Ribosomal protein L11b (Rpl11b), one of the ∼45 ribosomal proteins of the 60S subunit, was tagged at its carboxyl terminus with the green fluorescent protein (GFP) to enable visualization of the 60S subunit in living cells. A panel of mutant yeast strains was screened for their accumulation of Rpl11b–GFP in the nucleus as an indicator of their involvement in ribosome synthesis and/or transport. This panel included conditional alleles of several rRNA-processing factors, nucleoporins, general transport factors, and karyopherins. As predicted, conditional alleles of rRNA-processing factors that affect 60S ribosomal subunit assembly accumulated Rpl11b–GFP in the nucleus. In addition, several of the nucleoporin mutants as well as a few of the karyopherin and transport factor mutants also mislocalized Rpl11b–GFP. In particular, deletion of the previously uncharacterized karyopherin KAP120 caused accumulation of Rpl11b–GFP in the nucleus, whereas ribosomal protein import was not impaired. Together, these data further define the requirements for ribosomal subunit export and suggest a biological function for KAP120. PMID:11071906

Functional diversification of maize RNA polymerase IV and V subtypes via alternative catalytic subunits

DOE PAGES

Haag, Jeremy R.; Brower-Toland, Brent; Krieger, Elysia K.; ...

2014-10-02

Unlike nuclear multisubunit RNA polymerases I, II, and III, whose subunit compositions are conserved throughout eukaryotes, plant RNA polymerases IV and V are nonessential, Pol II-related enzymes whose subunit compositions are still evolving. Whereas Arabidopsis Pols IV and V differ from Pol II in four or five of their 12 subunits, respectively, and differ from one another in three subunits, proteomic analyses show that maize Pols IV and V differ from Pol II in six subunits but differ from each other only in their largest subunits. Use of alternative catalytic second subunits, which are nonredundant for development and paramutation, yieldsmore » at least two sub-types of Pol IV and three subtypes of Pol V in maize. Pol IV/Pol V associations with MOP1, RMR1, AGO121, Zm_DRD1/CHR127, SHH2a, and SHH2b extend parallels between paramutation in maize and the RNA-directed DNA methylation pathway in Arabidopsis.« less

Functional diversification of maize RNA polymerase IV and V subtypes via alternative catalytic subunits

PubMed Central

Haag, Jeremy R.; Brower-Toland, Brent; Krieger, Elysia K.; Sidorenko, Lyudmila; Nicora, Carrie D.; Norbeck, Angela D.; Irsigler, Andre; LaRue, Huachun; Brzeski, Jan; McGinnis, Karen; Ivashuta, Sergey; Pasa-Tolic, Ljiljana; Chandler, Vicki L.; Pikaard, Craig S.

2014-01-01

Summary Unlike nuclear multisubunit RNA polymerases I, II and III, whose subunit compositions are conserved throughout eukaryotes, plant RNA Polymerases IV and V are non-essential, Pol II-related enzymes whose subunit compositions are still evolving. Whereas Arabidopsis Pols IV and V differ from Pol II in four or five of their twelve subunits, respectively, and differ from one another in three subunits, proteomic analyses show that maize Pols IV and V differ from Pol II in six subunits, but differ from each other only in their largest subunits. Use of alternative catalytic second-subunits, which are non-redundant for development and paramutation, yields at least two subtypes of Pol IV, and three subtypes of Pol V in maize. Pol IV/V associations with MOP1, RMR1, AGO121, Zm_DRD1/CHR127, SHH2a and SHH2b extend parallels between paramutation in maize and the RNA-directed DNA methylation pathway in Arabidopsis. PMID:25284785

The 2.3 {angstrom} crystal structure of cholera toxin B subunit pentamer: Choleragenoid

DOE Office of Scientific and Technical Information (OSTI.GOV)

Zhang, Rong-Guang; Westbrook, M.L.; Maulik, P.R.

1996-02-01

Cholera toxin, a heterohexameric AB{sub 5} enterotoxin released by Vibrio cholera, induces a profuse secretory diarrhea in susceptible hosts. Choleragenoid, the B subunit pentamer of cholera toxin, directs the enzymatic A subunit to its target by binding to GM{sub 1} gangliosides exposed on the luminal surface of intestinal epithelial cells. We have solved the crystal structure of choleragenoid at 2.3 {Angstrom} resolution by combining single isomorphous replacement with non-crystallographic symmetry averaging. The structure of the B subunits, and their pentameric arrangement, closely resembles that reported for the intact holotoxin (choleragen), the heat-labile enterotoxin from E. coli, and for a choleragenoid-GM{submore » 1} pentasaccharide complex. In the absence of the A subunit the central cavity of the B pentamer is a highly solvated channel. The binding of the A subunit or the receptor pentasaccharide to choleragenoid has only a modest effect on the local stereochemistry and does not perceptibly alter the subunit interface.« less

Receptor-binding region in human choriogonadotropin/lutropin. beta. subunit

DOE Office of Scientific and Technical Information (OSTI.GOV)

Keutmann, H.T.; Charlesworth, M.C.; Mason, K.A.

1987-04-01

Synthetic fragments have not been widely used thus far to evaluate structure-activity relations in the glycoprotein hormones. The authors prepared a series of peptides representing the intercysteine loop sequence (residues 38-57) in human choriogonadotropin (hCG) and lutropin (hLH) ..beta.. subunits, anticipating that it might be oriented toward the surface and accessible to receptors. The peptides were characterized chemically and tested for bioactivity by binding to rat ovarian membrane receptor and stimulation of Leydig cell testosterone production. The hCG..beta..-(38-57) and hLH..beta..-(38-57) peptides inhibited binding of /sup 125/I-labeled hCG half-maximally at 1.51 x 10/sup -4/ and 2.03 x 10/sup -5/ M, respectively,more » while other peptide hormones and fragments from elsewhere in the ..beta.. subunit were inactive. Both peptides stimulated testosterone production, with half-maximal responses at 3.55 x 10/sup -5/ M (hCG) and 2.18 x 10/sup -5/ M (hLH). By radioimmunoassay with an antibody to thyroglobulin-conjugated hCG..beta..-(38-57) peptide, native hCG and ..beta.. subunit were highly reactive, as were the reduced and carboxymethylated subunit and peptide. These results indicate that the 38-57 region of ..beta.. subunit is exposed on the surface and constitutes a component in the receptor-binding domain for hCG and hLH. A region of amphipathic-helical structure in the 38-57 sequence may promote hormone-receptor interactions in a manner proposed for several other peptide hormones.« less

Vimentin filament precursors exchange subunits in an ATP-dependent manner

PubMed Central

Robert, Amélie; Rossow, Molly J.; Hookway, Caroline; Adam, Stephen A.; Gelfand, Vladimir I.

2015-01-01

Intermediate filaments (IFs) are a component of the cytoskeleton capable of profound reorganization in response to specific physiological situations, such as differentiation, cell division, and motility. Various mechanisms were proposed to be responsible for this plasticity depending on the type of IF polymer and the biological context. For example, recent studies suggest that mature vimentin IFs (VIFs) undergo rearrangement by severing and reannealing, but direct subunit exchange within the filament plays little role in filament dynamics at steady state. Here, we studied the dynamics of subunit exchange in VIF precursors, called unit-length filaments (ULFs), formed by the lateral association of eight vimentin tetramers. To block vimentin assembly at the ULF stage, we used the Y117L vimentin mutant (vimentinY117L). By tagging vimentinY117L with a photoconvertible protein mEos3.2 and photoconverting ULFs in a limited area of the cytoplasm, we found that ULFs, unlike mature filaments, were highly dynamic. Subunit exchange among ULFs occurred within seconds and was limited by the diffusion of soluble subunits in the cytoplasm rather than by the association and dissociation of subunits from ULFs. Our data demonstrate that cells expressing vimentinY117L contained a large pool of soluble vimentin tetramers that was in rapid equilibrium with ULFs. Furthermore, vimentin exchange in ULFs required ATP, and ATP depletion caused a dramatic reduction of the soluble tetramer pool. We believe that the dynamic exchange of subunits plays a role in the regulation of ULF assembly and the maintenance of a soluble vimentin pool during the reorganization of filament networks. PMID:26109569

Ribosomal protein L14 contributes to the early assembly of 60S ribosomal subunits in Saccharomyces cerevisiae.

PubMed

Espinar-Marchena, Francisco; Rodríguez-Galán, Olga; Fernández-Fernández, José; Linnemann, Jan; de la Cruz, Jesús

2018-05-18

The contribution of most ribosomal proteins to ribosome synthesis has been quite well analysed in Saccharomyces cerevisiae. However, few yeast ribosomal proteins still await characterization. Herein, we show that L14, an essential 60S ribosomal protein, assembles in the nucleolus at an early stage into pre-60S particles. Depletion of L14 results in a deficit in 60S subunits and defective processing of 27SA2 and 27SA3 to 27SB pre-rRNAs. As a result, 27S pre-rRNAs are subjected to turnover and export of pre-60S particles is blocked. These phenotypes likely appear as the direct consequence of the reduced pre-60S particle association not only of L14 upon its depletion but also of a set of neighboring ribosomal proteins located at the solvent interface of 60S subunits and the adjacent region surrounding the polypeptide exit tunnel. These pre-60S intermediates also lack some essential trans-acting factors required for 27SB pre-rRNA processing but accumulate practically all factors required for processing of 27SA3 pre-rRNA. We have also analysed the functional interaction between the eukaryote-specific carboxy-terminal extensions of the neighboring L14 and L16 proteins. Our results indicate that removal of the most distal parts of these extensions cause slight translation alterations in mature 60S subunits.

Communication Fact Sheet.

ERIC Educational Resources Information Center

American Speech-Language-Hearing Association, Rockville, MD.

This brief fact sheet examines key aspects of communication, communication disabilities, and intervention. The fact sheet addresses the following questions: the nature of communication; communication disabilities (definitions of hearing impairments and speech and language impairments are given); effects of communication disabilities (factors…

MspA Nanopores from Subunit Dimers

PubMed Central

Pavlenok, Mikhail; Derrington, Ian M.; Gundlach, Jens H.; Niederweis, Michael

2012-01-01

Mycobacterium smegmatis porin A (MspA) forms an octameric channel and represents the founding member of a new family of pore proteins. Control of subunit stoichiometry is important to tailor MspA for nanotechnological applications. In this study, two MspA monomers were connected by linkers ranging from 17 to 62 amino acids in length. The oligomeric pore proteins were purified from M. smegmatis and were shown to form functional channels in lipid bilayer experiments. These results indicated that the peptide linkers did not prohibit correct folding and localization of MspA. However, expression levels were reduced by 10-fold compared to wild-type MspA. MspA is ideal for nanopore sequencing due to its unique pore geometry and its robustness. To assess the usefulness of MspA made from dimeric subunits for DNA sequencing, we linked two M1-MspA monomers, whose constriction zones were modified to enable DNA translocation. Lipid bilayer experiments demonstrated that this construct also formed functional channels. Voltage gating of MspA pores made from M1 monomers and M1-M1 dimers was identical indicating similar structural and dynamic channel properties. Glucose uptake in M. smegmatis cells lacking porins was restored by expressing the dimeric mspA M1 gene indicating correct folding and localization of M1-M1 pores in their native membrane. Single-stranded DNA hairpins produced identical ionic current blockades in pores made from monomers and subunit dimers demonstrating that M1-M1 pores are suitable for DNA sequencing. This study provides the proof of principle that production of single-chain MspA pores in M. smegmatis is feasible and paves the way for generating MspA pores with altered stoichiometries. Subunit dimers enable better control of the chemical and physical properties of the constriction zone of MspA. This approach will be valuable both in understanding transport across the outer membrane in mycobacteria and in tailoring MspA for nanopore sequencing of DNA. PMID

Structural and molecular characterization of the prefoldin beta subunit from Thermococcus strain KS-1.

PubMed

Kida, Hiroshi; Sugano, Yuri; Iizuka, Ryo; Fujihashi, Masahiro; Yohda, Masafumi; Miki, Kunio

2008-11-14

Prefoldin (PFD) is a heterohexameric molecular chaperone that is found in eukaryotic cytosol and archaea. PFD is composed of alpha and beta subunits and forms a "jellyfish-like" structure. PFD binds and stabilizes nascent polypeptide chains and transfers them to group II chaperonins for completion of their folding. Recently, the whole genome of Thermococcus kodakaraensis KOD1 was reported and shown to contain the genes of two alpha and two beta subunits of PFD. The genome of Thermococcus strain KS-1 also possesses two sets of alpha (alpha1 and alpha2) and beta subunits (beta1 and beta2) of PFD (TsPFD). However, the functions and roles of each of these PFD subunits have not been investigated in detail. Here, we report the crystal structure of the TsPFD beta1 subunit at 1.9 A resolution and its functional analysis. TsPFD beta1 subunits form a tetramer with four coiled-coil tentacles resembling the jellyfish-like structure of heterohexameric PFD. The beta hairpin linkers of beta1 subunits assemble to form a beta barrel "body" around a central fourfold axis. Size-exclusion chromatography and multi-angle light-scattering analyses show that the beta1 subunits form a tetramer at pH 8.0 and a dimer of tetramers at pH 6.8. The tetrameric beta1 subunits can protect against aggregation of relatively small proteins, insulin or lysozyme. The structural and biochemical analyses imply that PFD beta1 subunits act as molecular chaperones in living cells of some archaea.

The Kv7.2/Kv7.3 heterotetramer assembles with a random subunit arrangement.

PubMed

Stewart, Andrew P; Gómez-Posada, Juan Camilo; McGeorge, Jessica; Rouhani, Maral J; Villarroel, Alvaro; Murrell-Lagnado, Ruth D; Edwardson, J Michael

2012-04-06

Voltage-gated K(+) channels composed of Kv7.2 and Kv7.3 are the predominant contributors to the M-current, which plays a key role in controlling neuronal activity. Various lines of evidence have indicated that Kv7.2 and Kv7.3 form a heteromeric channel. However, the subunit stoichiometry and arrangement within this putative heteromer are so far unknown. Here, we have addressed this question using atomic force microscopy imaging of complexes between isolated Kv7.2/Kv7.3 channels and antibodies to epitope tags on the two subunits, Myc on Kv7.2 and HA on Kv7.3. Initially, tsA 201 cells were transiently transfected with equal amounts of cDNA for the two subunits. The heteromer was isolated through binding of either tag to immunoaffinity beads and then decorated with antibodies to the other tag. In both cases, the distribution of angles between pairs of bound antibodies had two peaks, at around 90° and around 180°, and in both cases the 90° peak was about double the size of the 180° peak. These results indicate that the Kv7.2/Kv7.3 heteromer generated by cells expressing approximately equal amounts of the two subunits assembles as a tetramer with a predominantly 2:2 subunit stoichiometry and with a random subunit arrangement. When the DNA ratio for the two subunits was varied, copurification experiments indicated that the subunit stoichiometry was variable and not fixed at 2:2. Hence, there are no constraints on either the subunit stoichiometry or the subunit arrangement.

Lessons from an evolving rRNA: 16S and 23S rRNA structures from a comparative perspective

NASA Technical Reports Server (NTRS)

Gutell, R. R.; Larsen, N.; Woese, C. R.

1994-01-01

The 16S and 23S rRNA higher-order structures inferred from comparative analysis are now quite refined. The models presented here differ from their immediate predecessors only in minor detail. Thus, it is safe to assert that all of the standard secondary-structure elements in (prokaryotic) rRNAs have been identified, with approximately 90% of the individual base pairs in each molecule having independent comparative support, and that at least some of the tertiary interactions have been revealed. It is interesting to compare the rRNAs in this respect with tRNA, whose higher-order structure is known in detail from its crystal structure (36) (Table 2). It can be seen that rRNAs have as great a fraction of their sequence in established secondary-structure elements as does tRNA. However, the fact that the former show a much lower fraction of identified tertiary interactions and a greater fraction of unpaired nucleotides than the latter implies that many of the rRNA tertiary interactions remain to be located. (Alternatively, the ribosome might involve protein-rRNA rather than intramolecular rRNA interactions to stabilize three-dimensional structure.) Experimental studies on rRNA are consistent to a first approximation with the structures proposed here, confirming the basic assumption of comparative analysis, i.e., that bases whose compositions strictly covary are physically interacting. In the exhaustive study of Moazed et al. (45) on protection of the bases in the small-subunit rRNA against chemical modification, the vast majority of bases inferred to pair by covariation are found to be protected from chemical modification, both in isolated small-subunit rRNA and in the 30S subunit. The majority of the tertiary interactions are reflected in the chemical protection data as well (45). On the other hand, many of the bases not shown as paired in Fig. 1 are accessible to chemical attack (45). However, in this case a sizeable fraction of them are also protected against chemical

Purification of an eight subunit RNA polymerase I complex in Trypanosoma brucei.

PubMed

Nguyen, Tu N; Schimanski, Bernd; Zahn, André; Klumpp, Birgit; Günzl, Arthur

2006-09-01

Trypanosoma brucei harbors a unique multifunctional RNA polymerase (pol) I which transcribes, in addition to ribosomal RNA genes, the gene units encoding the major cell surface antigens variant surface glycoprotein and procyclin. In consequence, this RNA pol I is recruited to three structurally different types of promoters and sequestered to two distinct nuclear locations, namely the nucleolus and the expression site body. This versatility may require parasite-specific protein-protein interactions, subunits or subunit domains. Thus far, data mining of trypanosomatid genomes have revealed 13 potential RNA pol I subunits which include two paralogous sets of RPB5, RPB6, and RPB10. Here, we analyzed a cDNA library prepared from procyclic insect form T. brucei and found that all 13 candidate subunits are co-expressed. Moreover, we PTP-tagged the largest subunit TbRPA1, tandem affinity-purified the enzyme complex to homogeneity, and determined its subunit composition. In addition to the already known subunits RPA1, RPA2, RPC40, 1RPB5, and RPA12, the complex contained RPC19, RPB8, and 1RPB10. Finally, to evaluate the absence of RPB6 in our purifications, we used a combination of epitope-tagging and reciprocal coimmunoprecipitation to demonstrate that 1RPB6 but not 2RPB6 binds to RNA pol I albeit in an unstable manner. Collectively, our data strongly suggest that T. brucei RNA pol I binds a distinct set of the RPB5, RPB6, and RPB10 paralogs.

Sleep Apnea Facts

MedlinePlus

... Apnea Facts Sleep Apnea Links Sleep Apnea Facts Sleep apnea affects up to 18 million Americans The condition was ... member is the first to notice signs of sleep apnea in someone with the ... diagnosed. The condition affects about 4 percent of middle-aged men and ...

Fragrant Dioxane Derivatives Identify β1-Subunit-containing GABAA Receptors*

PubMed Central

Sergeeva, Olga A.; Kletke, Olaf; Kragler, Andrea; Poppek, Anja; Fleischer, Wiebke; Schubring, Stephan R.; Görg, Boris; Haas, Helmut L.; Zhu, Xin-Ran; Lübbert, Hermann; Gisselmann, Günter; Hatt, Hanns

2010-01-01

Nineteen GABAA receptor (GABAAR) subunits are known in mammals with only a restricted number of functionally identified native combinations. The physiological role of β1-subunit-containing GABAARs is unknown. Here we report the discovery of a new structural class of GABAAR positive modulators with unique β1-subunit selectivity: fragrant dioxane derivatives (FDD). At heterologously expressed α1βxγ2L (x-for 1,2,3) GABAAR FDD were 6 times more potent at β1- versus β2- and β3-containing receptors. Serine at position 265 was essential for the high sensitivity of the β1-subunit to FDD and the β1N286W mutation nearly abolished modulation; vice versa the mutation β3N265S shifted FDD sensitivity toward the β1-type. In posterior hypothalamic neurons controlling wakefulness GABA-mediated whole-cell responses and GABAergic synaptic currents were highly sensitive to FDD, in contrast to β1-negative cerebellar Purkinje neurons. Immunostaining for the β1-subunit and the potency of FDD to modulate GABA responses in cultured hypothalamic neurons was drastically diminished by β1-siRNA treatment. In conclusion, with the help of FDDs we reveal a functional expression of β1-containing GABAARs in the hypothalamus, offering a new tool for studies on the functional diversity of native GABAARs. PMID:20511229

28 CFR 51.6 - Political subunits.

Code of Federal Regulations, 2011 CFR

2011-07-01

... THE VOTING RIGHTS ACT OF 1965, AS AMENDED General Provisions § 51.6 Political subunits. All political... coverage by obtaining the declaratory judgment described in section 4(a) of the Act are subject to the...

7 CFR 275.7 - Selection of sub-units for review.

Code of Federal Regulations, 2014 CFR

2014-01-01

... situations, the sub-unit would be classified based upon its primary function exclusively. However, when any... classification and if selected for review, all functions performed shall be examined. For example, if a sub-unit... functions related to more than one of the areas. For example, coupon issuers must maintain a level of coupon...

7 CFR 275.7 - Selection of sub-units for review.

Code of Federal Regulations, 2011 CFR

2011-01-01

... situations, the sub-unit would be classified based upon its primary function exclusively. However, when any... classification and if selected for review, all functions performed shall be examined. For example, if a sub-unit... functions related to more than one of the areas. For example, coupon issuers must maintain a level of coupon...

7 CFR 275.7 - Selection of sub-units for review.

Code of Federal Regulations, 2012 CFR

2012-01-01

... situations, the sub-unit would be classified based upon its primary function exclusively. However, when any... classification and if selected for review, all functions performed shall be examined. For example, if a sub-unit... functions related to more than one of the areas. For example, coupon issuers must maintain a level of coupon...

7 CFR 275.7 - Selection of sub-units for review.

Code of Federal Regulations, 2013 CFR

2013-01-01

... situations, the sub-unit would be classified based upon its primary function exclusively. However, when any... classification and if selected for review, all functions performed shall be examined. For example, if a sub-unit... functions related to more than one of the areas. For example, coupon issuers must maintain a level of coupon...

The histone H2A deubiquitinase Usp16 regulates hematopoiesis and hematopoietic stem cell function.

PubMed

Gu, Yue; Jones, Amanda E; Yang, Wei; Liu, Shanrun; Dai, Qian; Liu, Yudong; Swindle, C Scott; Zhou, Dewang; Zhang, Zhuo; Ryan, Thomas M; Townes, Tim M; Klug, Christopher A; Chen, Dongquan; Wang, Hengbin

2016-01-05

Epigenetic mechanisms play important regulatory roles in hematopoiesis and hematopoietic stem cell (HSC) function. Subunits of polycomb repressive complex 1 (PRC1), the major histone H2A ubiquitin ligase, are critical for both normal and pathological hematopoiesis; however, it is unclear which of the several counteracting H2A deubiquitinases functions along with PRC1 to control H2A ubiquitination (ubH2A) level and regulates hematopoiesis in vivo. Here we investigated the function of Usp16 in mouse hematopoiesis. Conditional deletion of Usp16 in bone marrow resulted in a significant increase of global ubH2A level and lethality. Usp16 deletion did not change HSC number but was associated with a dramatic reduction of mature and progenitor cell populations, revealing a role in governing HSC lineage commitment. ChIP- and RNA-sequencing studies in HSC and progenitor cells revealed that Usp16 bound to many important hematopoietic regulators and that Usp16 deletion altered the expression of genes in transcription/chromosome organization, immune response, hematopoietic/lymphoid organ development, and myeloid/leukocyte differentiation. The altered gene expression was partly rescued by knockdown of PRC1 subunits, suggesting that Usp16 and PRC1 counterbalance each other to regulate cellular ubH2A level and gene expression in the hematopoietic system. We further discovered that knocking down Cdkn1a (p21cip1), a Usp16 target and regulated gene, rescued the altered cell cycle profile and differentiation defect of Usp16-deleted HSCs. Collectively, these studies identified Usp16 as one of the histone H2A deubiquitinases, which coordinates with the H2A ubiquitin ligase PRC1 to regulate hematopoiesis, and revealed cell cycle regulation by Usp16 as key for HSC differentiation.

22 CFR 16.8 - Agency review.

Code of Federal Regulations, 2010 CFR

2010-04-01

...), the Director of Personnel and Manpower (AID), and the Chief, Employee-Management Relations Division... correspondence under § 16.7(a), any documentary evidence readily available to the grievant on which the grievance... documentary evidence, and, in that official's discretion, interview persons having knowledge of the facts. The...

Vector dark energy and high-z massive clusters

NASA Astrophysics Data System (ADS)

Carlesi, Edoardo; Knebe, Alexander; Yepes, Gustavo; Gottlöber, Stefan; Jiménez, Jose Beltrán.; Maroto, Antonio L.

2011-12-01

The detection of extremely massive clusters at z > 1 such as SPT-CL J0546-5345, SPT-CL J2106-5844 and XMMU J2235.3-2557 has been considered by some authors as a challenge to the standard Λ cold dark matter cosmology. In fact, assuming Gaussian initial conditions, the theoretical expectation of detecting such objects is as low as ≤1 per cent. In this paper we discuss the probability of the existence of such objects in the light of the vector dark energy paradigm, showing by means of a series of N-body simulations that chances of detection are substantially enhanced in this non-standard framework.

Multiple alpha subunits of integrin are involved in cell-mediated responses of the Manduca immune system.

PubMed

Zhuang, Shufei; Kelo, Lisha; Nardi, James B; Kanost, Michael R

2008-01-01

The cell-mediated responses of the insect innate immune system-phagocytosis, nodulation, encapsulation-involve multiple cell adhesion molecules of hemocyte surfaces. A hemocyte-specific (HS) integrin and a member of the immunoglobulin (Ig) superfamily (neuroglian) are involved in the encapsulation response of hemocytes in Manduca sexta. In addition, two new integrin alpha (alpha) subunits have been found on these hemocytes. The alpha2 subunit is mainly expressed in epidermis and Malphigian tubules, whereas the alpha3 subunit is primarily expressed on hemocytes and fat body cells. Of the three known alpha subunits, the alpha1 subunit found in HS integrin is the predominant subunit of hemocytes. Cell adhesion assays indicate that alpha2 belongs to the integrin family with RGD-binding motifs, confirming the phylogenetic analysis of alpha subunits based on the amino-acid sequence alignment of different alpha subunits. Double-stranded RNAs (dsRNAs) targeting each of these three integrin alpha subunits not only specifically decreased transcript expression of each alpha subunit in hemocytes, but also abolished the cell-mediated encapsulation response of hemocytes to foreign surfaces. The individual alpha subunits of M. sexta integrins, like their integrin counterparts in mammalian immune systems, have critical, individual roles in cell-substrate and cell-cell interactions during immune responses.

Developmental expression of human hemoglobins mediated by maturation of their subunit interfaces

PubMed Central

Manning, Lois R; Popowicz, Anthony M; Padovan, Julio; Chait, Brian T; Russell, J Eric; Manning, James M

2010-01-01

Different types of human hemoglobins (Hbs) consisting of various combinations of the embryonic, fetal, and adult Hb subunits are present at certain times during development representing a major paradigm of developmental biology that is still not understood and one which we address here. We show that the subunit interfaces of these Hbs have increasing bonding strengths as demonstrated by their distinct distribution of tetramers, dimers, and monomers during gel filtration at very low-Hb concentration. This maturation is mediated by competition between subunits for more favorable partners with stronger subunit interactions. Thus, the protein products of gene expression can themselves have a role in the developmental process due to their intrinsic properties. PMID:20572018

Mining Protein Evolution for Insights into Mechanisms of Voltage-Dependent Sodium Channel Auxiliary Subunits.

PubMed

Molinarolo, Steven; Granata, Daniele; Carnevale, Vincenzo; Ahern, Christopher A

2018-02-21

Voltage-gated sodium channel (VGSC) beta (β) subunits have been called the "overachieving" auxiliary ion channel subunit. Indeed, these subunits regulate the trafficking of the sodium channel complex at the plasma membrane and simultaneously tune the voltage-dependent properties of the pore-forming alpha-subunit. It is now known that VGSC β-subunits are capable of similar modulation of multiple isoforms of related voltage-gated potassium channels, suggesting that their abilities extend into the broader voltage-gated channels. The gene family for these single transmembrane immunoglobulin beta-fold proteins extends well beyond the traditional VGSC β1-β4 subunit designation, with deep roots into the cell adhesion protein family and myelin-related proteins - where inherited mutations result in a myriad of electrical signaling disorders. Yet, very little is known about how VGSC β-subunits support protein trafficking pathways, the basis for their modulation of voltage-dependent gating, and, ultimately, their role in shaping neuronal excitability. An evolutionary approach can be useful in yielding new clues to such functions as it provides an unbiased assessment of protein residues, folds, and functions. An approach is described here which indicates the greater emergence of the modern β-subunits roughly 400 million years ago in the early neurons of Bilateria and bony fish, and the unexpected presence of distant homologues in bacteriophages. Recent structural breakthroughs containing α and β eukaryotic sodium channels containing subunits suggest a novel role for a highly conserved polar contact that occurs within the transmembrane segments. Overall, a mixture of approaches will ultimately advance our understanding of the mechanism for β-subunit interactions with voltage-sensor containing ion channels and membrane proteins.

20 Facts on Women Workers. Fact Sheet No. 88-2.

ERIC Educational Resources Information Center

Women's Bureau (DOL), Washington, DC.

This fact sheet lists 20 interpreted statistics on women workers. The facts cover the following data: number of women workers and their percentage in the labor force; length of time women are expected to stay in the labor force; racial and ethnic groups in the labor force; part-time and full-time employment; types of occupations in which women are…

Two distinctive β subunits are separately involved in two binding sites of imidacloprid with different affinities in Locusta migratoria manilensis.

PubMed

Bao, Haibo; Liu, Yang; Zhang, Yixi; Liu, Zewen

2017-08-01

Due to great diversity of nicotinic acetylcholine receptor (nAChR) subtypes in insects, one β subunit may be contained in numerous nAChR subtypes. In the locust Locusta migratoria, a model insect species with agricultural importance, the third β subunits (Locβ3) was identified in this study, which reveals at least three β subunits in this insect species. Imidacloprid was found to bind nAChRs in L. migratoria central nervous system at two sites with different affinities, with K d values of 0.16 and 10.31nM. The specific antisera (L1-1, L2-1 and L3-1) were raised against fusion proteins at the large cytoplasmic loop of Locβ1, Locβ2 and Locβ3 respectively. Specific immunodepletion of Locβ1 with antiserum L1-1 resulted in the selective loss of the low affinity binding site for imidacloprid, whereas the immunodepletion of Locβ3 with L3-1 caused the selective loss of the high affinity site. Dual immunodepletion with L1-1 and L3-1 could completely abolish imidacloprid binding. In contrast, the immunodepletion of Locβ2 had no significant effect on the specific [ 3 H]imidacloprid binding. Taken together, these data indicated that Locβ1 and Locβ3 were respectively contained in the low- and high-affinity binding sites for imidacloprid in L. migratoria, which is different to the previous finding in Nilaparvata lugens that Nlβ1 was in two binding sites for imidacloprid. The involvement of two β subunits separately in two binding sites may decrease the risk of imidacloprid resistance due to putative point mutations in β subunits in L. migratoria. Copyright © 2017 Elsevier B.V. All rights reserved.

Therapeutic potential of Mediator complex subunits in metabolic diseases.

PubMed

Ranjan, Amol; Ansari, Suraiya A

2018-01-01

The multisubunit Mediator is an evolutionary conserved transcriptional coregulatory complex in eukaryotes. It is needed for the transcriptional regulation of gene expression in general as well as in a gene specific manner. Mediator complex subunits interact with different transcription factors as well as components of RNA Pol II transcription initiation complex and in doing so act as a bridge between gene specific transcription factors and general Pol II transcription machinery. Specific interaction of various Mediator subunits with nuclear receptors (NRs) and other transcription factors involved in metabolism has been reported in different studies. Evidences indicate that ligand-activated NRs recruit Mediator complex for RNA Pol II-dependent gene transcription. These NRs have been explored as therapeutic targets in different metabolic diseases; however, they show side-effects as targets due to their overlapping involvement in different signaling pathways. Here we discuss the interaction of various Mediator subunits with transcription factors involved in metabolism and whether specific interaction of these transcription factors with Mediator subunits could be potentially utilized as therapeutic strategy in a variety of metabolic diseases. Copyright © 2017 Elsevier B.V. and Société Française de Biochimie et Biologie Moléculaire (SFBBM). All rights reserved.

Palmitoylation of the β4-Subunit Regulates Surface Expression of Large Conductance Calcium-activated Potassium Channel Splice Variants*

PubMed Central

Chen, Lie; Bi, Danlei; Tian, Lijun; McClafferty, Heather; Steeb, Franziska; Ruth, Peter; Knaus, Hans Guenther; Shipston, Michael J.

2013-01-01

Regulatory β-subunits of large conductance calcium- and voltage-activated potassium (BK) channels play an important role in generating functional diversity and control of cell surface expression of the pore forming α-subunits. However, in contrast to α-subunits, the role of reversible post-translational modification of intracellular residues on β-subunit function is largely unknown. Here we demonstrate that the human β4-subunit is S-acylated (palmitoylated) on a juxtamembrane cysteine residue (Cys-193) in the intracellular C terminus of the regulatory β-subunit. β4-Subunit palmitoylation is important for cell surface expression and endoplasmic reticulum (ER) exit of the β4-subunit alone. Importantly, palmitoylated β4-subunits promote the ER exit and surface expression of the pore-forming α-subunit, whereas β4-subunits that cannot be palmitoylated do not increase ER exit or surface expression of α-subunits. Strikingly, however, this palmitoylation- and β4-dependent enhancement of α-subunit surface expression was only observed in α-subunits that contain a putative trafficking motif (… REVEDEC) at the very C terminus of the α-subunit. Engineering this trafficking motif to other C-terminal α-subunit splice variants results in α-subunits with reduced surface expression that can be rescued by palmitoylated, but not depalmitoylated, β4-subunits. Our data reveal a novel mechanism by which palmitoylated β4-subunit controls surface expression of BK channels through masking of a trafficking motif in the C terminus of the α-subunit. As palmitoylation is dynamic, this mechanism would allow precise control of specific splice variants to the cell surface. Our data provide new insights into how complex interplay between the repertoire of post-transcriptional and post-translational mechanisms controls cell surface expression of BK channels. PMID:23504458

Facts about Hib Disease. ARC Facts.

ERIC Educational Resources Information Center

Association for Retarded Citizens, Arlington, TX.

The fact sheet provides basic information about Hib Disease in young children, which may involve a bacterial meningitis causing mental retardation, hearing loss, partial blindness, speech disorders, partial paralysis, behavioral problems, or seizures. Stressed is prevention of Hib Disease through immunization. The question and answer format…

Modulation of Gain-of-function α6*-Nicotinic Acetylcholine Receptor by β3 Subunits*

PubMed Central

Dash, Bhagirathi; Lukas, Ronald J.

2012-01-01

We previously have shown that β3 subunits either eliminate (e.g. for all-human (h) or all-mouse (m) α6β4β3-nAChR) or potentiate (e.g. for hybrid mα6hβ4hβ3- or mα6mβ4hβ3-nAChR containing subunits from different species) function of α6*-nAChR expressed in Xenopus oocytes, and that nAChR hα6 subunit residues Asn-143 and Met-145 in N-terminal domain loop E are important for dominant-negative effects of nAChR hβ3 subunits on hα6*-nAChR function. Here, we tested the hypothesis that these effects of β3 subunits would be preserved even if nAChR α6 subunits harbored gain-of-function, leucine- or valine-to-serine mutations at 9′ or 13′ positions (L9′S or V13′S) in their second transmembrane domains, yielding receptors with heightened functional activity and more amenable to assessment of effects of β3 subunit incorporation. However, coexpression with β3 subunits potentiates rather than suppresses function of all-human, all-mouse, or hybrid α6(L9′S or V13′S)β4*- or α6(N143D+M145V)L9′Sβ2*-nAChR. This contrasts with the lack of consistent function when α6(L9′S or V13′S) and β2 subunits are expressed alone or in the presence of wild-type β3 subunits. These results provide evidence that gain-of-function hα6hβ2*-nAChR (i.e. hα6(N143D+M145V)L9′Shβ2hβ3 nAChR) could be produced in vitro. These studies also indicate that nAChR β3 subunits can be assembly partners in functional α6*-nAChR and that 9′ or 13′ mutations in the nAChR α6 subunit second transmembrane domain can act as gain-of-function and/or reporter mutations. Moreover, our findings suggest that β3 subunit coexpression promotes function of α6*-nAChR. PMID:22315221

The cytochrome oxidase subunit I and subunit III genes in Oenothera mitochondria are transcribed from identical promoter sequences

PubMed Central

Hiesel, Rudolf; Schobel, Werner; Schuster, Wolfgang; Brennicke, Axel

1987-01-01

Two loci encoding subunit III of the cytochrome oxidase (COX) in Oenothera mitochondria have been identified from a cDNA library of mitochondrial transcripts. A 657-bp sequence block upstream from the open reading frame is also present in the two copies of the COX subunit I gene and is presumably involved in homologous sequence rearrangement. The proximal points of sequence rearrangements are located 3 bp upstream from the COX I and 1139 bp upstream from the COX III initiation codons. The 5'-termini of both COX I and COX III mRNAs have been mapped in this common sequence confining the promoter region for the Oenothera mitochondrial COX I and COX III genes to the homologous sequence block. ImagesFig. 5. PMID:15981332

Electrophysiological and biochemical evidence that DEG/ENaC cation channels are composed of nine subunits.

PubMed

Snyder, P M; Cheng, C; Prince, L S; Rogers, J C; Welsh, M J

1998-01-09

Members of the DEG/ENaC protein family form ion channels with diverse functions. DEG/ENaC subunits associate as hetero- and homomultimers to generate channels; however the stoichiometry of these complexes is unknown. To determine the subunit stoichiometry of the human epithelial Na+ channel (hENaC), we expressed the three wild-type hENaC subunits (alpha, beta, and gamma) with subunits containing mutations that alter channel inhibition by methanethiosulfonates. The data indicate that hENaC contains three alpha, three beta, and three gamma subunits. Sucrose gradient sedimentation of alphahENaC translated in vitro, as well as alpha-, beta-, and gammahENaC coexpressed in cells, was consistent with complexes containing nine subunits. FaNaCh and BNC1, two related DEG/ENaC channels, produced complexes of similar mass. Our results suggest a novel nine-subunit stoichiometry for the DEG/ENaC family of ion channels.

Antigen discovery and delivery of subunit vaccines by nonliving bacterial ghost vectors.

PubMed

Walcher, Petra; Mayr, Ulrike B; Azimpour-Tabrizi, Chakameh; Eko, Francis O; Jechlinger, Wolfgang; Mayrhofer, Peter; Alefantis, Tim; Mujer, Cesar V; DelVecchio, Vito G; Lubitz, Werner

2004-12-01

The bacterial ghost (BG) platform system is a novel vaccine delivery system endowed with intrinsic adjuvant properties. BGs are nonliving Gram-negative bacterial cell envelopes which are devoid of their cytoplasmic contents, yet maintain their cellular morphology and antigenic structures, including bioadhesive properties. The main advantages of BGs as carriers of subunit vaccines include their ability to stimulate a high immune response and to target the carrier itself to primary antigen-presenting cells. The intrinsic adjuvant properties of BGs enhance the immune response to target antigens, including T-cell activation and mucosal immunity. Since native and foreign antigens can be carried in the envelope complex of BGs, combination vaccines with multiple antigens of diverse origin can be presented to the immune system simultaneously. Beside the capacity of BGs to function as carriers of protein antigens, they also have a high loading capacity for DNA. Thus, loading BGs with recombinant DNA takes advantage of the excellent bioavailability for DNA-based vaccines and the high expression rates of the DNA-encoded antigens in target cell types such as macrophages and dendritic cells. There are many spaces within BGs including the inner and outer membranes, the periplasmic space and the internal lumen which can carry antigens, DNA or mediators of the immune response. All can be used for subunit antigen to design new vaccine candidates with particle presentation technology. In addition, the fact that BGs can also carry piggyback large-size foreign antigen particles, increases the technologic usefulness of BGs as combination vaccines against viral and bacterial pathogens. Furthermore, the BG antigen carriers can be stored as freeze-dried preparations at room temperature for extended periods without loss of efficacy. The potency, safety and relatively low production cost of BGs offer a significant technical advantage over currently utilized vaccine technologies.

Atypical properties of a conventional calcium channel beta subunit from the platyhelminth Schistosoma mansoni.

PubMed

Salvador-Recatalà, Vicenta; Schneider, Toni; Greenberg, Robert M

2008-03-26

The function of voltage-gated calcium (Cav) channels greatly depends on coupling to cytoplasmic accessory beta subunits, which not only promote surface expression, but also modulate gating and kinetic properties of the alpha1 subunit. Schistosomes, parasitic platyhelminths that cause schistosomiasis, express two beta subunit subtypes: a structurally conventional beta subunit and a variant beta subunit with unusual functional properties. We have previously characterized the functional properties of the variant Cavbeta subunit. Here, we focus on the modulatory phenotype of the conventional Cavbeta subunit (SmCavbeta) using the human Cav2.3 channel as the substrate for SmCavbeta and the whole-cell patch-clamp technique. The conventional Schistosoma mansoni Cavbeta subunit markedly increases Cav2.3 currents, slows macroscopic inactivation and shifts steady state inactivation in the hyperpolarizing direction. However, currents produced by Cav2.3 in the presence of SmCavbeta run-down to approximately 75% of their initial amplitudes within two minutes of establishing the whole-cell configuration. This suppressive effect was independent of Ca2+, but dependent on intracellular Mg2+-ATP. Additional experiments revealed that SmCavbeta lends the Cav2.3/SmCavbeta complex sensitivity to Na+ ions. A mutant version of the Cavbeta subunit lacking the first forty-six amino acids, including a string of twenty-two acidic residues, no longer conferred sensitivity to intracellular Mg2+-ATP and Na+ ions, while continuing to show wild type modulation of current amplitude and inactivation of Cav2.3. The data presented in this article provide insights into novel mechanisms employed by platyhelminth Cavbeta subunits to modulate voltage-gated Ca2+ currents that indicate interactions between the Ca2+ channel complex and chelated forms of ATP as well as Na+ ions. These results have potentially important implications for understanding previously unknown mechanisms by which platyhelminths and

Fragrant dioxane derivatives identify beta1-subunit-containing GABAA receptors.

PubMed

Sergeeva, Olga A; Kletke, Olaf; Kragler, Andrea; Poppek, Anja; Fleischer, Wiebke; Schubring, Stephan R; Görg, Boris; Haas, Helmut L; Zhu, Xin-Ran; Lübbert, Hermann; Gisselmann, Günter; Hatt, Hanns

2010-07-30

Nineteen GABA(A) receptor (GABA(A)R) subunits are known in mammals with only a restricted number of functionally identified native combinations. The physiological role of beta1-subunit-containing GABA(A)Rs is unknown. Here we report the discovery of a new structural class of GABA(A)R positive modulators with unique beta1-subunit selectivity: fragrant dioxane derivatives (FDD). At heterologously expressed alpha1betaxgamma2L (x-for 1,2,3) GABA(A)R FDD were 6 times more potent at beta1- versus beta2- and beta3-containing receptors. Serine at position 265 was essential for the high sensitivity of the beta1-subunit to FDD and the beta1N286W mutation nearly abolished modulation; vice versa the mutation beta3N265S shifted FDD sensitivity toward the beta1-type. In posterior hypothalamic neurons controlling wakefulness GABA-mediated whole-cell responses and GABAergic synaptic currents were highly sensitive to FDD, in contrast to beta1-negative cerebellar Purkinje neurons. Immunostaining for the beta1-subunit and the potency of FDD to modulate GABA responses in cultured hypothalamic neurons was drastically diminished by beta1-siRNA treatment. In conclusion, with the help of FDDs we reveal a functional expression of beta1-containing GABA(A)Rs in the hypothalamus, offering a new tool for studies on the functional diversity of native GABA(A)Rs.

Molecular Cloning of Pituitary Glycoprotein α-Subunit and Follicle Stimulating Hormone and Chorionic Gonadotropin β-Subunits from New World Squirrel Monkey and Owl Monkey

PubMed Central

Scammell, Jonathan G.; Funkhouser, Jane D.; Moyer, Felricia S.; Gibson, Susan V.; Willis, Donna L.

2008-01-01

The goal of this study was to characterize the gonadotropins expressed in pituitary glands of the New World squirrel monkey (Saimiri sp.) and owl monkey (Aotus sp.). The various subunits were amplified from total RNA from squirrel monkey and owl monkey pituitary glands by reverse transcription-polymerase chain reaction and the deduced amino acid sequences compared to those of other species. Mature squirrel monkey and owl monkey glycoprotein hormone α-polypeptides (96 amino acids in length) were determined to be 80% homologous to the human sequence. The sequences of mature β subunits of follicle stimulating hormone (FSHβ) from squirrel monkey and owl monkey (111 amino acids in length) are 92% homologous to human FSHβ. New World primate glycoprotein hormone α-polypeptides and FSHβ subunits showed conservation of all cysteine residues and consensus N-linked glycosylation sites. Attempts to amplify the β-subunit of luteinizing hormone from squirrel monkey and owl monkey pituitary glands were unsuccessful. Rather, the β-subunit of chorionic gonadotropin (CG) was amplified from pituitaries of both New World primates. Squirrel monkey and owl monkey CGβ are 143 and 144 amino acids in length and 77% homologous with human CGβ. The greatest divergence is in the C terminus, where all four sites for O-linked glycosylation in human CGβ, responsible for delayed metabolic clearance, are predicted to be absent in New World primate CGβs. It is likely that CG secreted from pituitary of New World primates exhibits a relatively short half-life compared to human CG. PMID:17897645

Molecular cloning of pituitary glycoprotein alpha-subunit and follicle stimulating hormone and chorionic gonadotropin beta-subunits from New World squirrel monkey and owl monkey.

PubMed

Scammell, Jonathan G; Funkhouser, Jane D; Moyer, Felricia S; Gibson, Susan V; Willis, Donna L

2008-02-01

The goal of this study was to characterize the gonadotropins expressed in pituitary glands of the New World squirrel monkey (Saimiri sp.) and owl monkey (Aotus sp.). The various subunits were amplified from total RNA from squirrel monkey and owl monkey pituitary glands by reverse transcription-polymerase chain reaction and the deduced amino acid sequences compared to those of other species. Mature squirrel monkey and owl monkey glycoprotein hormone alpha-polypeptides (96 amino acids in length) were determined to be 80% homologous to the human sequence. The sequences of mature beta subunits of follicle stimulating hormone (FSHbeta) from squirrel monkey and owl monkey (111 amino acids in length) are 92% homologous to human FSHbeta. New World primate glycoprotein hormone alpha-polypeptides and FSHbeta subunits showed conservation of all cysteine residues and consensus N-linked glycosylation sites. Attempts to amplify the beta-subunit of luteinizing hormone from squirrel monkey and owl monkey pituitary glands were unsuccessful. Rather, the beta-subunit of chorionic gonadotropin (CG) was amplified from pituitaries of both New World primates. Squirrel monkey and owl monkey CGbeta are 143 and 144 amino acids in length and 77% homologous with human CGbeta. The greatest divergence is in the C terminus, where all four sites for O-linked glycosylation in human CGbeta, responsible for delayed metabolic clearance, are predicted to be absent in New World primate CGbetas. It is likely that CG secreted from pituitary of New World primates exhibits a relatively short half-life compared to human CG.

Copolymer semiconductors comprising thiazolothiazole or benzobisthiazole, or benzobisoxazole electron acceptor subunits, and electron donor subunits, and their uses in transistors and solar cells

DOEpatents

Jenekhe, Samson A; Subramaniyan, Selvam; Ahmed, Eilaf; Xin, Hao; Kim, Felix Sunjoo

2014-10-28

The inventions disclosed, described, and/or claimed herein relate to copolymers comprising copolymers comprising electron accepting A subunits that comprise thiazolothiazole, benzobisthiazole, or benzobisoxazoles rings, and electron donating subunits that comprise certain heterocyclic groups. The copolymers are useful for manufacturing organic electronic devices, including transistors and solar cells. The invention also relates to certain synthetic precursors of the copolymers. Methods for making the copolymers and the derivative electronic devices are also described.

The NMDA receptor NR2A subunit regulates proliferation of MKN45 human gastric cancer cells

DOE Office of Scientific and Technical Information (OSTI.GOV)

Watanabe, Kanako; Department of Anesthesiology, Hyogo College of Medicine, 1-1 Mukogawa-cho, Nishinomiya 663-8501; Kanno, Takeshi

2008-03-07

The present study investigated proliferation of MKN28 and MKN45 human gastric cancer cells regulated by the N-methyl-D-aspartate (NMDA) receptor subunit. The NMDA receptor antagonist DL-2-amino-5-phosphonovaleric acid (AP5) inhibited proliferation of MKN45 cells, but not MKN28 cells. Of the NMDA subunits such as NR1, NR2 (2A, 2B, 2C, and 2D), and NR3 (3A and 3B), all the NMDA subunit mRNAs except for the NR2B subunit mRNA were expressed in both MKN28 and MKN45 cells. MKN45 cells were characterized by higher expression of the NR2A subunit mRNA and lower expression of the NR1 subunit mRNA, but MKN28 otherwise by higher expression ofmore » the NR1 subunit mRNA and lower expression of the NR2A subunit mRNA. MKN45 cell proliferation was also inhibited by silencing the NR2A subunit-targeted gene. For MKN45 cells, AP5 or knocking-down the NR2A subunit increased the proportion of cells in the G{sub 1} phase of cell cycling and decreased the proportion in the S/G{sub 2} phase. The results of the present study, thus, suggest that blockage of NMDA receptors including the NR2A subunit suppresses MKN45 cell proliferation due to cell cycle arrest at the G{sub 1} phase; in other words, the NR2A subunit promotes MKN45 cell proliferation by accelerating cell cycling.« less

Immunocytochemical localization of the NMDA-R2A receptor subunit in the cat retina.

PubMed

Goebel, D J; Aurelia, J L; Tai, Q; Jojich, L; Poosch, M S

1998-10-19

Immunocytochemical studies were performed to determine the distribution and cellular localization of the NMDA-R2A receptor subunit (R2A) in the cat retina. R2A-immunoreactivity (R2A-IR) was noted in all layers of the retina, with specific localizations in the outer segments of red/green and blue cone photoreceptors, B-type horizontal cells, several types of amacrine cells, Müller cells and the majority of cells in the ganglion cell layer. In the inner nuclear layer, 48% of all cells residing in the amacrine cell layer were R2A-IR including a cell resembling the GABAergic A17 amacrine cell. Interestingly, the AII rod amacrine cell was devoid of R2A-IR. Although the localization of the R2A subunit was anticipated in ganglion cells, amacrines and Müller cells, the presence of this receptor subunit to the cells in the outer retina was not expected. Here, both the R2A and the R2B subunits were found to be present in the outer segments of cone photoreceptors and to the tips of rod outer segments. Although the function of these receptor subunits in rod and cone photoreceptors remains to be determined, the fact that both R2A and R2B receptor subunits are localized to cone outer segments suggests a possible alternative pathway for calcium entry into a region where this cation plays such a crucial role in the process of phototransduction. To further classify the cells that display NR2A-IR, we performed dual labeling experiments showing the relationship between R2A-labeled cells with GABA. Results showed that all GABAergic-amacrines and displaced amacrines express the R2A-subunit protein. In addition, approximately 11% of the NR2A-labeled amacrines, did not stain for GABA. These findings support pharmacological data showing that NMDA directly facilitates GABA release in retina and retinal cultures [I.L. Ferreira, C.B. Duarte, P.F. Santos, C.M. Carvalho, A.P. Carvalho, Release of [3H]GABA evoked by glutamate receptor agonist in cultured chick retinal cells: effect of Ca2

AFM study of Escherichia coli RNA polymerase σ⁷⁰ subunit aggregation.

PubMed

Dubrovin, Evgeniy V; Koroleva, Olga N; Khodak, Yulia A; Kuzmina, Natalia V; Yaminsky, Igor V; Drutsa, Valeriy L

2012-01-01

The self-assembly of Escherichia coli RNA polymerase σ⁷⁰ subunit was investigated using several experimental approaches. A novel rodlike shape was reported for σ⁷⁰ subunit aggregates. Atomic force microscopy reveals that these aggregates, or σ⁷⁰ polymers, have a straight rodlike shape 5.4 nm in diameter and up to 300 nm in length. Atomic force microscopy data, Congo red binding assay, and sodium dodecyl sulfate gel electrophoresis confirm the amyloid nature of observed aggregates. The process of formation of rodlike structures proceeds spontaneously under nearly physiological conditions. E. coli RNA polymerase σ⁷⁰ subunit may be an interesting object for investigation of amyloidosis as well as for biotechnological applications that exploit self-assembled bionanostructures. Polymerization of σ⁷⁰ subunit may be a competitive process with its three-dimensional crystallization and association with core RNA polymerase. In this basic science study, the self-assembly of Escherichia coli RNA polymerase σ⁷⁰( subunit was investigated using atomic force microscopy and other complementary approaches. 2012 Elsevier Inc. All rights reserved.

Fluorescence detection of the movement of single KcsA subunits reveals cooperativity

PubMed Central

Blunck, Rikard; McGuire, Hugo; Hyde, H. Clark; Bezanilla, Francisco

2008-01-01

The prokaryotic KcsA channel is gated at the helical bundle crossing by intracellular protons and inactivates at the extracellular selectivity filter. The C-terminal transmembrane helix has to undergo a conformational change for potassium ions to access the central cavity. Whereas a partial opening of the tetrameric channel is suggested to be responsible for subconductance levels of ion channels, including KcsA, a cooperative opening of the 4 subunits is postulated as the final opening step. In this study, we used single-channel fluorescence spectroscopy of KcsA to directly observe the movement of each subunit and the temporal correlation between subunits. Purified KcsA channels labeled at the C terminus near the bundle crossing have been inserted into supported lipid bilayer, and the fluorescence traces analyzed by means of a cooperative or independent Markov model. The analysis revealed that the 4 subunits do not move fully independently but instead showed a certain degree of cooperativity. However, the 4 subunits do not simply open in 1 concerted step. PMID:19074286

Supportive periodontal therapy and periodontal biotype as prognostic factors in implants placed in patients with a history of periodontitis.

PubMed

Aguirre-Zorzano, Luis-Antonio; Vallejo-Aisa, Francisco-Javier; Estefanía-Fresco, Ruth

2013-09-01

To evaluate bone loss around implants placed in patients with a history of treated chronic periodontitis and who did or did not attend supportive periodontal therapy, after one year in function. Furthermore, the influence of periodontal biotype and level of plaque was also evaluated. Forty-nine patients participated voluntarily in the study. All subjects had a history of chronic periodontitis, which had been previously treated. After the active treatment, 27 patients attended supportive periodontal therapy (SPT) and the rest did not (No SPT). The O'Leary plaque index and periodontal biotype were recorded for each subject and 246 Astra Tech® OsseospeedTM implants were radiographically analysed (123 placed in SPT patients and 123 in No SPT patients) at the time of loading and one year later, measuring marginal bone loss with the program Dental Studio NX 6.0®. The statistical analysis was performed with Windows SPSS, applying Pearson's correlation index and the Kruskal-Wallis and U-Mann Whitney non-parametric tests. Six patients were found to have periimplantitis and sixteen mucositis. The survival rate was 99.59% (100% SPT and 99.18% No SPT). Mean bone loss was 0.39 mm (range [-0.71 - 8.05]). Among SPT patients, 95% of the implants had losses less than or equal to the mean (mean bone loss of 0.16 mm) compared to 53.7% for the No SPT group (mean bone loss of 0.62 mm). A statistically significant relationship was demonstrated between bone loss around the implant and the patient's periodontal biotype and plaque index. The marginal bone loss around implants in patients with treated chronic periodontitis is minimal if they are in a controlled SPT programme and there is individual control of plaque index. Moreover, the presence of a thin periodontal biotype represents a risk factor for additional bone loss.

45 CFR 681.16 - Can the reviewing official or ALJ be disqualified?

Code of Federal Regulations, 2014 CFR

2014-10-01

... SCIENCE FOUNDATION PROGRAM FRAUD CIVIL REMEDIES ACT REGULATIONS Hearing Procedures § 681.16 Can the... follows: (1) The motion must be supported by an affidavit containing specific facts establishing that... discovery of such facts; (2) The motion must be filed promptly after discovery of the grounds for...

45 CFR 681.16 - Can the reviewing official or ALJ be disqualified?

Code of Federal Regulations, 2011 CFR

2011-10-01

... SCIENCE FOUNDATION PROGRAM FRAUD CIVIL REMEDIES ACT REGULATIONS Hearing Procedures § 681.16 Can the... follows: (1) The motion must be supported by an affidavit containing specific facts establishing that... discovery of such facts; (2) The motion must be filed promptly after discovery of the grounds for...

45 CFR 681.16 - Can the reviewing official or ALJ be disqualified?

Code of Federal Regulations, 2012 CFR

2012-10-01

... SCIENCE FOUNDATION PROGRAM FRAUD CIVIL REMEDIES ACT REGULATIONS Hearing Procedures § 681.16 Can the... follows: (1) The motion must be supported by an affidavit containing specific facts establishing that... discovery of such facts; (2) The motion must be filed promptly after discovery of the grounds for...

45 CFR 681.16 - Can the reviewing official or ALJ be disqualified?

Code of Federal Regulations, 2010 CFR

2010-10-01

... SCIENCE FOUNDATION PROGRAM FRAUD CIVIL REMEDIES ACT REGULATIONS Hearing Procedures § 681.16 Can the... follows: (1) The motion must be supported by an affidavit containing specific facts establishing that... discovery of such facts; (2) The motion must be filed promptly after discovery of the grounds for...

45 CFR 681.16 - Can the reviewing official or ALJ be disqualified?

Code of Federal Regulations, 2013 CFR

2013-10-01

... SCIENCE FOUNDATION PROGRAM FRAUD CIVIL REMEDIES ACT REGULATIONS Hearing Procedures § 681.16 Can the... follows: (1) The motion must be supported by an affidavit containing specific facts establishing that... discovery of such facts; (2) The motion must be filed promptly after discovery of the grounds for...

Cholera Toxin B: One Subunit with Many Pharmaceutical Applications

PubMed Central

Baldauf, Keegan J.; Royal, Joshua M.; Hamorsky, Krystal Teasley; Matoba, Nobuyuki

2015-01-01

Cholera, a waterborne acute diarrheal disease caused by Vibrio cholerae, remains prevalent in underdeveloped countries and is a serious health threat to those living in unsanitary conditions. The major virulence factor is cholera toxin (CT), which consists of two subunits: the A subunit (CTA) and the B subunit (CTB). CTB is a 55 kD homopentameric, non-toxic protein binding to the GM1 ganglioside on mammalian cells with high affinity. Currently, recombinantly produced CTB is used as a component of an internationally licensed oral cholera vaccine, as the protein induces potent humoral immunity that can neutralize CT in the gut. Additionally, recent studies have revealed that CTB administration leads to the induction of anti-inflammatory mechanisms in vivo. This review will cover the potential of CTB as an immunomodulatory and anti-inflammatory agent. We will also summarize various recombinant expression systems available for recombinant CTB bioproduction. PMID:25802972

Protein Kinase A Subunit Balance Regulates Lipid Metabolism in Caenorhabditis elegans and Mammalian Adipocytes*

PubMed Central

Lee, Jung Hyun; Han, Ji Seul; Kong, Jinuk; Ji, Yul; Lv, Xuchao; Lee, Junho; Li, Peng; Kim, Jae Bum

2016-01-01

Protein kinase A (PKA) is a cyclic AMP (cAMP)-dependent protein kinase composed of catalytic and regulatory subunits and involved in various physiological phenomena, including lipid metabolism. Here we demonstrated that the stoichiometric balance between catalytic and regulatory subunits is crucial for maintaining basal PKA activity and lipid homeostasis. To uncover the potential roles of each PKA subunit, Caenorhabditis elegans was used to investigate the effects of PKA subunit deficiency. In worms, suppression of PKA via RNAi resulted in severe phenotypes, including shortened life span, decreased egg laying, reduced locomotion, and altered lipid distribution. Similarly, in mammalian adipocytes, suppression of PKA regulatory subunits RIα and RIIβ via siRNAs potently stimulated PKA activity, leading to potentiated lipolysis without increasing cAMP levels. Nevertheless, insulin exerted anti-lipolytic effects and restored lipid droplet integrity by antagonizing PKA action. Together, these data implicate the importance of subunit stoichiometry as another regulatory mechanism of PKA activity and lipid metabolism. PMID:27496951

The ribosomal subunit assembly line

PubMed Central

Dlakić, Mensur

2005-01-01

Recent proteomic studies in Saccharomyces cerevisiae have identified nearly 200 proteins, other than the structural ribosomal proteins, that participate in the assembly of ribosomal subunits and their transport from the nucleus. In a separate line of research, proteomic studies of mature plant ribosomes have revealed considerable variability in the protein composition of individual ribosomes. PMID:16207363

Subunit architecture and functional modular rearrangements of the transcriptional mediator complex.

PubMed

Tsai, Kuang-Lei; Tomomori-Sato, Chieri; Sato, Shigeo; Conaway, Ronald C; Conaway, Joan W; Asturias, Francisco J

2014-06-05

The multisubunit Mediator, comprising ∼30 distinct proteins, plays an essential role in gene expression regulation by acting as a bridge between DNA-binding transcription factors and the RNA polymerase II (RNAPII) transcription machinery. Efforts to uncover the Mediator mechanism have been hindered by a poor understanding of its structure, subunit organization, and conformational rearrangements. By overcoming biochemical and image analysis hurdles, we obtained accurate EM structures of yeast and human Mediators. Subunit localization experiments, docking of partial X-ray structures, and biochemical analyses resulted in comprehensive mapping of yeast Mediator subunits and a complete reinterpretation of our previous Mediator organization model. Large-scale Mediator rearrangements depend on changes at the interfaces between previously described Mediator modules, which appear to be facilitated by factors conducive to transcription initiation. Conservation across eukaryotes of Mediator structure, subunit organization, and RNA polymerase II interaction suggest conservation of fundamental aspects of the Mediator mechanism. Copyright © 2014 Elsevier Inc. All rights reserved.

Subunit architecture and functional modular rearrangements of the transcriptional Mediator complex

PubMed Central

Tsai, Kuang-Lei; Tomomori-Sato, Chieri; Sato, Shigeo; Conaway, Ronald C.; Conaway, Joan W.; Asturias, Francisco J.

2014-01-01

SUMMARY The multisubunit Mediator comprising ~30 distinct proteins, plays an essential role in gene expression regulation by acting as a bridge between DNA binding transcription factors and the RNA polymerase II (RNAPII) transcription machinery. Efforts to uncover the Mediator mechanism have been hindered by a poor understanding of its structure, subunit organization, and conformational rearrangements. By overcoming biochemical and image analysis hurdles, we obtained accurate EM structures of yeast and human Mediators. Subunit localization experiments, docking of partial X-ray structures, and biochemical analyses resulted in comprehensive mapping of yeast Mediator subunits and a complete reinterpretation of our previous Mediator organization model. Large-scale Mediator rearrangements depend on changes at the interfaces between previously described Mediator modules, which appear to be facilitated by factors conducive to transcription initiation. Conservation across eukaryotes of Mediator structure, subunit organization, and RNA polymerase II interaction suggest conservation of fundamental aspects of the Mediator mechanism. PMID:24882805

Crash survival analysis of 16 agricultural aircraft accidents.

DOT National Transportation Integrated Search

1972-04-01

The study presents pertinent findings from on-the-scene investigations to evaluate the crashworthiness of the present fleet of agricultural applicator aircraft. A detailed presentation of 16 crashes illustrates the fact that most of these specialized...

Ribosomal protein L14 contributes to the early assembly of 60S ribosomal subunits in Saccharomyces cerevisiae

PubMed Central

Espinar-Marchena, Francisco; Rodríguez-Galán, Olga; Fernández-Fernández, José; Linnemann, Jan; de la Cruz, Jesús

2018-01-01

Abstract The contribution of most ribosomal proteins to ribosome synthesis has been quite well analysed in Saccharomyces cerevisiae. However, few yeast ribosomal proteins still await characterization. Herein, we show that L14, an essential 60S ribosomal protein, assembles in the nucleolus at an early stage into pre-60S particles. Depletion of L14 results in a deficit in 60S subunits and defective processing of 27SA2 and 27SA3 to 27SB pre-rRNAs. As a result, 27S pre-rRNAs are subjected to turnover and export of pre-60S particles is blocked. These phenotypes likely appear as the direct consequence of the reduced pre-60S particle association not only of L14 upon its depletion but also of a set of neighboring ribosomal proteins located at the solvent interface of 60S subunits and the adjacent region surrounding the polypeptide exit tunnel. These pre-60S intermediates also lack some essential trans-acting factors required for 27SB pre-rRNA processing but accumulate practically all factors required for processing of 27SA3 pre-rRNA. We have also analysed the functional interaction between the eukaryote-specific carboxy-terminal extensions of the neighboring L14 and L16 proteins. Our results indicate that removal of the most distal parts of these extensions cause slight translation alterations in mature 60S subunits. PMID:29788267

Recent Advances in Subunit Vaccine Carriers

PubMed Central

Vartak, Abhishek; Sucheck, Steven J.

2016-01-01

The lower immunogenicity of synthetic subunit antigens, compared to live attenuated vaccines, is being addressed with improved vaccine carriers. Recent reports indicate that the physio-chemical properties of these carriers can be altered to achieve optimal antigen presentation, endosomal escape, particle bio-distribution, and cellular trafficking. The carriers can be modified with various antigens and ligands for dendritic cells targeting. They can also be modified with adjuvants, either covalently or entrapped in the matrix, to improve cellular and humoral immune responses against the antigen. As a result, these multi-functional carrier systems are being explored for use in active immunotherapy against cancer and infectious diseases. Advancing technology, improved analytical methods, and use of computational methodology have also contributed to the development of subunit vaccine carriers. This review details recent breakthroughs in the design of nano-particulate vaccine carriers, including liposomes, polymeric nanoparticles, and inorganic nanoparticles. PMID:27104575

Modulation of BK channel voltage gating by different auxiliary β subunits

PubMed Central

Contreras, Gustavo F.; Neely, Alan; Alvarez, Osvaldo; Gonzalez, Carlos; Latorre, Ramon

2012-01-01

Calcium- and voltage-activated potassium channels (BK) are regulated by a multiplicity of signals. The prevailing view is that different BK gating mechanisms converge to determine channel opening and that these gating mechanisms are allosterically coupled. In most instances the pore forming α subunit of BK is associated with one of four alternative β subunits that appear to target specific gating mechanisms to regulate the channel activity. In particular, β1 stabilizes the active configuration of the BK voltage sensor having a large effect on BK Ca2+ sensitivity. To determine the extent to which β subunits regulate the BK voltage sensor, we measured gating currents induced by the pore-forming BK α subunit alone and with the different β subunits expressed in Xenopus oocytes (β1, β2IR, β3b, and β4). We found that β1, β2, and β4 stabilize the BK voltage sensor in the active conformation. β3 has no effect on voltage sensor equilibrium. In addition, β4 decreases the apparent number of charges per voltage sensor. The decrease in the charge associated with the voltage sensor in α β4 channels explains most of their biophysical properties. For channels composed of the α subunit alone, gating charge increases slowly with pulse duration as expected if a significant fraction of this charge develops with a time course comparable to that of K+ current activation. In the presence of β1, β2, and β4 this slow component develops in advance of and much more rapidly than ion current activation, suggesting that BK channel opening proceeds in two steps. PMID:23112204

Interaction of KCNE subunits with the KCNQ1 K+ channel pore

PubMed Central

Panaghie, Gianina; Tai, Kwok-Keung; Abbott, Geoffrey W

2006-01-01

KCNQ1 α subunits form functionally distinct potassium channels by coassembling with KCNE ancillary subunits MinK and MiRP2. MinK-KCNQ1 channels generate the slowly activating, voltage-dependent cardiac IKs current. MiRP2-KCNQ1 channels form a constitutively active current in the colon. The structural basis for these contrasting channel properties, and the mechanisms of α subunit modulation by KCNE subunits, are not fully understood. Here, scanning mutagenesis located a tryptophan-tolerant region at positions 338–340 within the KCNQ1 pore-lining S6 domain, suggesting an exposed region possibly amenable to interaction with transmembrane ancillary subunits. This hypothesis was tested using concomitant mutagenesis in KCNQ1 and in the membrane-localized ‘activation triplet’ regions of MinK and MiRP2 to identify pairs of residues that interact to control KCNQ1 activation. Three pairs of mutations exerted dramatic effects, ablating channel function or either removing or restoring control of KCNQ1 activation. The results place KCNE subunits close to the KCNQ1 pore, indicating interaction of MiRP2-72 with KCNQ1-338; and MinK-59,58 with KCNQ1-339, 340. These data are consistent either with perturbation of the S6 domain by MinK or MiRP2, dissimilar positioning of MinK and MiRP2 within the channel complex, or both. Further, the results suggest specifically that two of the interactions, MiRP2-72/KCNQ1-338 and MinK-58/KCNQ1-340, are required for the contrasting gating effects of MinK and MiRP2. PMID:16308347

Fine structure of OXI1, the mitochondrial gene coding for subunit II of yeast cytochrome c oxidase.

PubMed

Weiss-Brummer, B; Guba, R; Haid, A; Schweyen, R J

1979-12-01

Genetic and biochemical studies have been performed with 110 mutants which are defective in cytochrome a·a3 and map in the regions on mit DNA previously designated OXI1 and OXI2. With 88 mutations allocated to OXI1 fine structure mapping was achieved by the analysis of rho (-) deletions. The order of six groups of mutational sites (A 1, A2, B 1, B2, C 1, C2) thus determined was confirmed by oxi i x oxi j recombination analysis.Analysis of mitochondrially translated polypeptides of oxil mutants by SDS-polyacrylamide electrophoresis reveals three classes of mutant patterns: i) similar to wild-tpye (19 mutants); ii) lacking SU II of cytochrome c oxidase (53 mutants); iii) lacking this subunit and exhibiting a single new polypeptide of lower Mr (16 mutants). Mutations of each of these classes are scattered over the OXI1 region without any detectable clustering; this is consistent with the assumption that all oxil mutations studied are within the same gene.New polypeptides observed in oxil mutants of class iii) vary in Mr in the range from 10,500 to 33,000. Those of Mr 17,000 to 33,000 are shown to be antigenically related to subunit II of cytochrome c oxidase. Colinearity is established between the series of new polypeptides of Mr values increasing from 10,500 to 31,500 and the order of the respective mutational sites on the map, e.g. mutations mapping in A 1 generate the smallest and mutations mapping in C2 the largest mutant fragments.From these data we conclude that i) all mutations allocated to the OXI1 region are in the same gene; ii) this gene codes for subunit II of cytochrome c oxidase; iii) the direction of translation is from CAP to 0X12. Out of 19 mutants allocated to OXI2 three exhibit a new polypeptide; these and all the other oxi2 mutants lack subunit III of cytochrome oxidase. This result provides preliminary evidence that the OXI2 region harbours the structural gene for this subunit III.

Subunits of the Drosophila Actin-Capping Protein Heterodimer Regulate Each Other at Multiple Levels

PubMed Central

Amândio, Ana Rita; Gaspar, Pedro; Whited, Jessica L.; Janody, Florence

2014-01-01

The actin-Capping Protein heterodimer, composed of the α and β subunits, is a master F-actin regulator. In addition to its role in many cellular processes, Capping Protein acts as a main tumor suppressor module in Drosophila and in humans, in part, by restricting the activity of Yorkie/YAP/TAZ oncogenes. We aimed in this report to understand how both subunits regulate each other in vivo. We show that the levels and capping activities of both subunits must be tightly regulated to control F-actin levels and consequently growth of the Drosophila wing. Overexpressing capping protein α and β decreases both F-actin levels and tissue growth, while expressing forms of Capping Protein that have dominant negative effects on F-actin promote tissue growth. Both subunits regulate each other's protein levels. In addition, overexpressing one of the subunit in tissues knocked-down for the other increases the mRNA and protein levels of the subunit knocked-down and compensates for its loss. We propose that the ability of the α and β subunits to control each other's levels assures that a pool of functional heterodimer is produced in sufficient quantities to restrict the development of tumor but not in excess to sustain normal tissue growth. PMID:24788460

Small things considered: the small accessory subunits of RNA polymerase in Gram-positive bacteria

PubMed Central

Weiss, Andy; Shaw, Lindsey N.

2015-01-01

The DNA-dependent RNA polymerase core enzyme in Gram-positive bacteria consists of seven subunits. Whilst four of them (α2ββ′) are essential, three smaller subunits, δ, ε and ω (∼9–21.5 kDa), are considered accessory. Both δ and ω have been viewed as integral components of RNAP for several decades; however, ε has only recently been described. Functionally these three small subunits carry out a variety of tasks, imparting important, supportive effects on the transcriptional process of Gram-positive bacteria. While ω is thought to have a wide range of roles, reaching from maintaining structural integrity of RNAP to σ factor recruitment, the only suggested function for ε thus far is in protecting cells from phage infection. The third subunit, δ, has been shown to have distinct influences in maintaining transcriptional specificity, and thus has a key role in cellular fitness. Collectively, all three accessory subunits, although dispensable under laboratory conditions, are often thought to be crucial for proper RNAP function. Herein we provide an overview of the available literature on each subunit, summarizing landmark findings that have deepened our understanding of these proteins and their function, and outline future challenges in understanding the role of these small subunits in the transcriptional process. PMID:25878038

Both α and β Subunits of Human Choriogonadotropin Photoaffinity Label the Hormone Receptor

NASA Astrophysics Data System (ADS)

Ji, Inhae; Ji, Tae H.

1981-09-01

It has been shown that a photoactivable derivative of human choriogonadotropin (hCG) labels the lutropin receptor on porcine granulosa cells [Ji, I. & Ji, T. H. (1980) Proc. Natl. Acad. Sci. USA 77, 7167-7170]. In an attempt to identify which of the hCG subunits labeled the receptor, three sets of different hCG derivatives were prepared. In the first set, hCG was coupled to the N-hydroxysuccinimide ester of 4-azidobenzoylglycine and radioiodinated. In the second set, only one of the subunits was radioiodinated, but both subunits were allowed to react with the reagent. In the third set, both the reagent and [125I]iodine were coupled to only one of the subunits. The binding activity of each hormone derivative was comparable to that of 125I-labeled hCG. After binding of these hormone derivatives to the granulosa cell surface, they were photolyzed. After solubilization, autoradiographs of sodium dodecyl sulfate/polyacrylamide gels of each sample revealed a number of labeled bands; the hCG derivatives containing 125I-labeled alpha subunit produced four bands (molecular weights 120,000 +/- 6,000, 96,000 +/- 5,000, 76,000 +/- 4,000, and 73,000 +/- 4,000) and those containing 125I-labeled beta subunit produced three bands (molecular weights 106,000 +/- 6,000, 88,000 +/- 5,000, and 83,000 +/- 4,000). Results were the same when the hormone-receptor complexes were solubilized in 0.5% Triton X-100 and then photolyzed or when the hormone was derivatized with a family of reagents having arms of various lengths. We conclude that both the alpha subunit and the beta subunit of hCG photoaffinity labeled certain membrane polypeptides and that these polypeptides are related to the hormone receptor.

Topology of subunits of the mammalian cytochrome c oxidase: Relationship to the assembly of the enzyme complex

DOE Office of Scientific and Technical Information (OSTI.GOV)

Yu-Zhong Zhang; Ewart, G.; Capaldi, R.A.

The arrangement of three subunits of beef heart cytochrome c oxidase, subunits Va, VIa, and VIII, has been explored by chemical labeling and protease digestion studies. Subunit Va is an extrinsic protein located on the C side of the mitochondrial inner membrane. This subunit was found to label with N-(4-azido-2-nitrophenyl)-2-aminoethane({sup 35}S)sulfonate and sodium methyl 4-({sup 3}H)formylphenyl phosphate in reconstituted vesicles in which 90% of cytochrome c oxidase complexes were oriented with the C domain outermost. Subunit VIa was cleaved by trypsin both in these reconstituted vesicles and in submitochondrial particles, indicating a transmembrane orientation. The epitope for a monoclonal antibodymore » (mAb) to subunit VIa was lost or destroyed when cleavage occurred in reconstituted vesicles. This epitope was localized to the C-terminal part of the subunit by antibody binding to a fusion protein consisting of glutathione S-transferase (G-ST) and the C-terminal amino acids 55-85 of subunit VIa. No antibody binding was obtained with a fusion protein containing G-ST and the N-terminal amino acids 1-55. The mAb reaction orients subunit VIa with its C-terminus in the C-domain. Subunit VIII was cleaved by trypsin in submitochondrial particles but not in reconstituted vesicles. N-Terminal sequencing of the subunit VIII cleavage produce from submitochondrial particles gave the same sequence as the untreated subunit, i.e., ITA, indicating that it is the C-terminus which is cleaved from the M side. Subunits Va and VIII each contain N-terminal extensions or leader sequences in the precursor polypeptides; subunit VIa is made without an N-terminal extension.« less

Glycosylation of β2 Subunits Regulates GABAA Receptor Biogenesis and Channel Gating*

PubMed Central

Lo, Wen-yi; Lagrange, Andre H.; Hernandez, Ciria C.; Harrison, Rebecca; Dell, Anne; Haslam, Stuart M.; Sheehan, Jonathan H.; Macdonald, Robert L.

2010-01-01

γ-Aminobutyric acid type A (GABAA) receptors are heteropentameric glycoproteins. Based on consensus sequences, the GABAA receptor β2 subunit contains three potential N-linked glycosylation sites, Asn-32, Asn-104, and Asn-173. Homology modeling indicates that Asn-32 and Asn-104 are located before the α1 helix and in loop L3, respectively, near the top of the subunit-subunit interface on the minus side, and that Asn-173 is located in the Cys-loop near the bottom of the subunit N-terminal domain. Using site-directed mutagenesis, we demonstrated that all predicted β2 subunit glycosylation sites were glycosylated in transfected HEK293T cells. Glycosylation of each site, however, produced specific changes in α1β2 receptor surface expression and function. Although glycosylation of Asn-173 in the Cys-loop was important for stability of β2 subunits when expressed alone, results obtained with flow cytometry, brefeldin A treatment, and endo-β-N-acetylglucosaminidase H digestion suggested that glycosylation of Asn-104 was required for efficient α1β2 receptor assembly and/or stability in the endoplasmic reticulum. Patch clamp recording revealed that mutation of each site to prevent glycosylation decreased peak α1β2 receptor current amplitudes and altered the gating properties of α1β2 receptor channels by reducing mean open time due to a reduction in the proportion of long open states. In addition to functional heterogeneity, endo-β-N-acetylglucosaminidase H digestion and glycomic profiling revealed that surface β2 subunit N-glycans at Asn-173 were high mannose forms that were different from those of Asn-32 and N104. Using a homology model of the pentameric extracellular domain of α1β2 channel, we propose mechanisms for regulation of GABAA receptors by glycosylation. PMID:20639197

Two subunits of the 55 K porcine zona pellucida glycoprotein family are immunologically distinct

DOE Office of Scientific and Technical Information (OSTI.GOV)

Subramanian, M.G.; Yurewicz, E.C.; Sacco, A.G.

1986-03-01

The 55K glycoprotein family (ZP3) of the porcine zona pellucida is comprised of two subunits of 46 K and 45 K which can be resolved by endo-..beta..-galactosidase digestion of ZP3 followed by reversed phase HPLC on Vydac C4 resin. Gel electrophoresis revealed that the 46 K component (EBDG..cap alpha..) is approx. 95% pure and the 45 K component (EBGD..beta..) is 100% pure. In the present study, these two subunits were evaluated immunologically by RIA. Under similar reaction protocols (chloramine-T iodination procedure) comparable specific activities were obtained for EBGD..cap alpha.. (33.06 +/- 7.5 ..mu..ci/..mu..gm), EBGD..beta.. (30.45 +/- 1.6) and ZP3 (26.3more » +/- 1.3). Antibody (Ab) titration studies revealed that EBGD..cap alpha.. and ..beta.. are potent immunogens and /sup 125/I-EBGD..cap alpha.. showed minimal cross reactivity to EBGD..beta..-Ab (8% bound at 1:500 dilution), whereas, /sup 125/I-EBGD..beta.. showed a greater degree of cross reactivity to EBGD..cap alpha..-Ab (23% bound at 1:500 dilution). Maximum binding for the two labeled antigens against homologous Abs (1:500) was > 60%. Dose response studies revealed that in the /sup 125/I-EBGD..cap alpha.. vs EBGD..cap alpha.. -Ab system, the 50% intercept was 3.25 +/- 0.32 ng for EBGD..cap alpha.. and 472.43 +/- 30.26 ng for EBGD..beta.. (p < 0.01), whereas, in the /sup 125/I-EBGD..beta.. vs EBGD..beta..-Ab system the 50% intercept was 3.51 +/- 0.58 for EBGD..beta.. and 166.77 +/- 49.20 for EBGD..cap alpha.. (p < 0.01). No significant differences were observed in the slopes of the dose response curves. It is concluded that the two subunits of ZP3 possess distinct immunologic characteristics as evaluated by RIA.« less

Meth (Crank, Ice) Facts

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... Crank, Ice) Facts Meth (Crank, Ice) Facts Listen Methamphetamine—meth for short—is a white, bitter powder. ... names for meth are: Crank Ice Crystal Glass Chalk In This Section Signs of Meth Use and ...

Key Facts about Tularemia

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... Methyl isocyanate Case Definition: Methyl Isocyanate Poisoning Mustard gas (H) (sulfur mustard) Facts About Sulfur Mustard Case ... choking/lung agents Ricin Riot control agents/tear gas Facts About Riot Control Agents Case Definition: Riot ...

Two subunits of human ORC are dispensable for DNA replication and proliferation.

PubMed

Shibata, Etsuko; Kiran, Manjari; Shibata, Yoshiyuki; Singh, Samarendra; Kiran, Shashi; Dutta, Anindya

2016-12-01

The six-subunit Origin Recognition Complex (ORC) is believed to be an essential eukaryotic ATPase that binds to origins of replication as a ring-shaped heterohexamer to load MCM2-7 and initiate DNA replication. We have discovered that human cell lines in culture proliferate with intact chromosomal origins of replication after disruption of both alleles of ORC2 or of the ATPase subunit, ORC1 . The ORC1 or ORC2 -depleted cells replicate with decreased chromatin loading of MCM2-7 and become critically dependent on another ATPase, CDC6, for survival and DNA replication. Thus, either the ORC ring lacking a subunit, even its ATPase subunit, can load enough MCM2-7 in partnership with CDC6 to initiate DNA replication, or cells have an ORC-independent, CDC6-dependent mechanism to load MCM2-7 on origins of replication.

ε Subunit of Bacillus subtilis F1-ATPase Relieves MgADP Inhibition

PubMed Central

Mizumoto, Junya; Kikuchi, Yuka; Nakanishi, Yo-Hei; Mouri, Naoto; Cai, Anrong; Ohta, Tokushiro; Haruyama, Takamitsu; Kato-Yamada, Yasuyuki

2013-01-01

MgADP inhibition, which is considered as a part of the regulatory system of ATP synthase, is a well-known process common to all F1-ATPases, a soluble component of ATP synthase. The entrapment of inhibitory MgADP at catalytic sites terminates catalysis. Regulation by the ε subunit is a common mechanism among F1-ATPases from bacteria and plants. The relationship between these two forms of regulatory mechanisms is obscure because it is difficult to distinguish which is active at a particular moment. Here, using F1-ATPase from Bacillus subtilis (BF1), which is strongly affected by MgADP inhibition, we can distinguish MgADP inhibition from regulation by the ε subunit. The ε subunit did not inhibit but activated BF1. We conclude that the ε subunit relieves BF1 from MgADP inhibition. PMID:23967352

Specific roles for the Ccr4-Not complex subunits in expression of the genome

PubMed Central

Azzouz, Nowel; Panasenko, Olesya O.; Deluen, Cécile; Hsieh, Julien; Theiler, Grégory; Collart, Martine A.

2009-01-01

In this work we used micro-array experiments to determine the role of each nonessential subunit of the conserved Ccr4-Not complex in the control of gene expression in the yeast Saccharomyces cerevisiae. The study was performed with cells growing exponentially in high glucose and with cells grown to glucose depletion. Specific patterns of gene deregulation were observed upon deletion of any given subunit, revealing the specificity of each subunit's function. Consistently, the purification of the Ccr4-Not complex through Caf40p by tandem affinity purification from wild-type cells or cells lacking individual subunits of the Ccr4-Not complex revealed that each subunit had a particular impact on complex integrity. Furthermore, the micro-arrays revealed that the role of each subunit was specific to the growth conditions. From the study of only two different growth conditions, revealing an impact of the Ccr4-Not complex on more than 85% of all studied genes, we can infer that the Ccr4-Not complex is important for expression of most of the yeast genome. PMID:19155328

Cocaine (Coke, Crack) Facts

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... That People Abuse » Cocaine (Coke, Crack) Facts Cocaine (Coke, Crack) Facts Listen Cocaine is a white ... 69 KB) "My life was built around getting cocaine and getting high." ©istock.com/ Marjot Stacey is ...

A kinetic comparison of the processing and secretion of the alpha beta dimer and the uncombined alpha and beta subunits of chorionic gonadotropin synthesized by human choriocarcinoma cells.

PubMed

Peters, B P; Krzesicki, R F; Hartle, R J; Perini, F; Ruddon, R W

1984-12-25

Human choriocarcinoma cells (JAR) synthesize the alpha and beta subunits of the glycoprotein hormone chorionic gonadotropin (hCG) (R.W. Ruddon, C.A. Hanson, A. H. Bryan, G.J. Putterman, E.L. White, F. Perini, K. S. Meade, and P.H. Aldenderfer (1980) J. Biol. Chem. 255, 1000-1007). In addition to the hCG dimer (alpha beta), JAR cells secrete uncombined alpha and beta subunits into the culture medium (L.A. Cole, R.J. Hartle, J.A. Laferla, and R.W. Ruddon (1983) Endocrinology 113, 1176-1178). Pulse-chase studies with [35S]methionine or [3H]mannose were carried out in order to compare free alpha, free beta, and the alpha beta dimer with regard to the kinetics of synthesis, N-linked oligosaccharide processing, and secretion and to determine the kinetics of alpha-beta subunit combination. A panel of three antisera was used to immunoprecipitate directly the free subunits and the alpha beta dimer sequentially from the same cell lysates and culture media. The alpha subunit of hCG was synthesized in a slight molar excess (1.2-1.5-fold) over the beta subunit, and alpha beta dimer was rapidly formed by combination of the intracellular alpha and beta precursors. Dimer formation was already apparent in JAR cells following a 10-min biosynthetic labeling incubation with [35S]methionine. The combination of subunits ceased by 30 min of chase even though 51% of alpha and 44% of beta remained free within the cells. Combination of the alpha and beta precursors had occurred before their N-linked oligosaccharides were processed beyond the Man8GlcNAc2 structure. The initial trimming of glucosyl and mannosyl units from the high-mannose oligosaccharides of the hCG precursors occurred more rapidly for free alpha and CG-alpha than for free beta and CG-beta. JAR cells accumulated alpha precursors bearing mostly Man8GlcNAc2 units and beta precursors bearing Man8GlcNAc2 units that represent the substrates of the rate-limiting step in the secretory pathway. In spite of the fact that their N

Heroin (Smack, Junk) Facts

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... That People Abuse » Heroin (Smack, Junk) Facts Heroin (Smack, Junk) Facts Listen ©istock.com/ Bestfotostudio Heroin ... 301.56 KB) (301.56 KB) "I needed heroin just to get by." ©istock.com/ pixelheadphoto Deon ...

Marijuana: Facts for Teens

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... Publications » Marijuana: Facts for Teens » Letter to Teens Marijuana: Facts for Teens Email Facebook Twitter Letter to ... about 1 in 14 teens say they used marijuana in the past month. 1 So, if you ...

Asymmetric configurations in a reengineered homodimer reveal multiple subunit communication pathways in protein allostery

PubMed Central

Lanfranco, Maria Fe; Gárate, Fernanda; Engdahl, Ashton J.; Maillard, Rodrigo A.

2017-01-01

Many allosteric proteins form homo-oligomeric complexes to regulate a biological function. In homo-oligomers, subunits establish communication pathways that are modulated by external stimuli like ligand binding. A challenge for dissecting the communication mechanisms in homo-oligomers is identifying intermediate liganded states, which are typically transiently populated. However, their identities provide the most mechanistic information on how ligand-induced signals propagate from bound to empty subunits. Here, we dissected the directionality and magnitude of subunit communication in a reengineered single-chain version of the homodimeric transcription factor cAMP receptor protein. By combining wild-type and mutant subunits in various asymmetric configurations, we revealed a linear relationship between the magnitude of cooperative effects and the number of mutant subunits. We found that a single mutation is sufficient to change the global allosteric behavior of the dimer even when one subunit was wild type. Dimers harboring two mutations with opposite cooperative effects had different allosteric properties depending on the arrangement of the mutations. When the two mutations were placed in the same subunit, the resulting cooperativity was neutral. In contrast, when placed in different subunits, the observed cooperativity was dominated by the mutation with strongest effects over cAMP affinity relative to wild type. These results highlight the distinct roles of intrasubunit interactions and intersubunit communication in allostery. Finally, dimers bound to either one or two cAMP molecules had similar DNA affinities, indicating that both asymmetric and symmetric liganded states activate DNA interactions. These studies have revealed the multiple communication pathways that homo-oligomers employ to transduce signals. PMID:28188293

Effects of the α subunit on imidacloprid sensitivity of recombinant nicotinic acetylcholine receptors

PubMed Central

Matsuda, K; Buckingham, S D; Freeman, J C; Squire, M D; Baylis, H A; Sattelle, D B

1998-01-01

Imidacloprid is a new insecticide with selective toxicity for insects over vertebrates. Recombinant (α4β2) chicken neuronal nicotinic acetylcholine receptors (AChRs) and a hybrid nicotinic AChR formed by co-expression of a Drosophila melanogaster neuronal α subunit (SAD) with the chicken β2 subunit were heterologously expressed in Xenopus oocytes by nuclear injection of cDNAs. The agonist actions of imidacloprid and other nicotinic AChR ligands ((+)-epibatidine, (−)-nicotine and acetylcholine) were compared on both recombinant nicotinic AChRs by use of two-electrode, voltage-clamp electrophysiology. Imidacloprid alone of the 4 agonists behaved as a partial agonist on the α4β2 receptor; (+)-epibatidine, (−)-nicotine and acetylcholine were all full, or near full, agonists. Imidacloprid was also a partial agonist of the hybrid Drosophila SAD chicken β2 receptor, as was (−)-nicotine, whereas (+)-epibatidine and acetylcholine were full agonists. The EC50 of imidacloprid was decreased by replacing the chicken α4 subunit with the Drosophila SAD α subunit. This α subunit substitution also resulted in an increase in the EC50 for (+)-epibatidine, (−)-nicotine and acetylcholine. Thus, the Drosophila (SAD) α subunit contributes to the greater apparent affinity of imidacloprid for recombinant insect/vertebrate nicotinic AChRs. Imidacloprid acted as a weak antagonist of ACh-mediated responses mediated by SADβ2 hybrid receptors and as a weak potentiator of ACh responses mediated by α4β2 receptors. This suggests that imidacloprid has complex effects upon these recombinant receptors, determined at least in part by the α subunit. PMID:9504393

Role of the HoxZ subunit in the electron transfer pathway of the membrane-bound [NiFe]-hydrogenase from Ralstonia eutropha immobilized on electrodes.

PubMed

Sezer, Murat; Frielingsdorf, Stefan; Millo, Diego; Heidary, Nina; Utesch, Tillman; Mroginski, Maria-Andrea; Friedrich, Bärbel; Hildebrandt, Peter; Zebger, Ingo; Weidinger, Inez M

2011-09-01

The role of the diheme cytochrome b (HoxZ) subunit in the electron transfer pathway of the membrane-bound [NiFe]-hydrogenase (MBH) heterotrimer from Ralstonia eutropha H16 has been investigated. The MBH in its native heterotrimeric state was immobilized on electrodes and subjected to spectroscopic and electrochemical analysis. Surface enhanced resonance Raman spectroscopy was used to monitor the redox and coordination state of the HoxZ heme cofactors while concomitant protein film voltammetric measurements gave insights into the catalytic response of the enzyme on the electrode. The entire MBH heterotrimer as well as its isolated HoxZ subunit were immobilized on silver electrodes coated with self-assembled monolayers of ω-functionalized alkylthiols, displaying the preservation of the native heme pocket structure and an electrical communication between HoxZ and the electrode. For the immobilized MBH heterotrimer, catalytic reduction of the HoxZ heme cofactors was observed upon H(2) addition. The catalytic currents of MBH with and without the HoxZ subunit were measured and compared with the heterogeneous electron transfer rates of the isolated HoxZ. On the basis of the spectroscopic and electrochemical results, we conclude that the HoxZ subunit under these artificial conditions is not primarily involved in the electron transfer to the electrode but plays a crucial role in stabilizing the enzyme on the electrode. © 2011 American Chemical Society

Cystic fibrosis transmembrane conductance regulator gene mutations: do they play a role in the aetiology of allergic bronchopulmonary aspergillosis?

PubMed

Eaton, T E; Weiner Miller, P; Garrett, J E; Cutting, G R

2002-05-01

Previous work suggests that cystic fibrosis transmembrane conductance regulator (CFTR) gene mutations may be implicated in the aetiology of allergic bronchopulmonary aspergilosis (ABPA). To compare the frequency of CF gene mutations in asthmatics with ABPA of varying severity with asthmatics who were skin prick test (SPT)-positive to Aspergillus fumigatus (Af) without evidence of ABPA and asthmatics SPT-negative to Af. Thirty-one Caucasian patients with ABPA were identified, together with asthmatics SPT positive to Af without evidence of ABPA (n = 23) and SPT negative to Af (n = 28). Genomic DNA was tested for 16 CF mutations accounting for approximately 85% of CF alleles in Caucasian New Zealanders. Four (12.9%) ABPA patients were found to be carriers of a CF mutation (DeltaF508 n = 3, R117H n = 1), one (4.3%) asthmatic SPT positive to Af without ABPA (DeltaF508), and one (3.6%) asthmatic SPT negative to Af (R117H). All patients with a CF mutation had normal sweat chloride (< 40 mM). There was no significant difference between the frequency of CF mutations in the ABPA patients and asthmatics without ABPA. However, the frequency of CF mutations in the ABPA patients was significantly different (P = 0.0125) to the expected carrier rate in the general population. These results lend further support to a possible link between CF mutations and ABPA.

Marijuana (Weed, Pot) Facts

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... That People Abuse » Marijuana (Weed, Pot) Facts Marijuana (Weed, Pot) Facts Listen Marijuana is a green, brown, or gray mix of dried, shredded leaves and flowers from the marijuana plant. Marijuana can be rolled up and smoked ...

Chemical crosslinking of the subunits of HIV-1 reverse transcriptase.

PubMed Central

Debyser, Z.; De Clercq, E.

1996-01-01

The reverse transcriptase (RT) of the human immunodeficiency virus type 1 (HIV-1) is composed of two subunits of 66 and 51 kDa in a 1 to 1 ratio. Because dimerization is a prerequisite for enzymatic activity, interference with the dimerization process could constitute an alternative antiviral strategy for RT inhibition. Here we describe an in vitro assay for the study of the dimerization state of HIV-1 reverse transcriptase based on chemical crosslinking of the subunits with dimethylsuberimidate. Crosslinking results in the formation of covalent bonds between the subunits, so that the crosslinked species can be resolved by denaturing gel electrophoresis. Crosslinked RT species with molecular weight greater than that of the dimeric form accumulate during a 1-15-min time course. Initial evidence suggests that those high molecular weight species represent trimers and tetramers and may be the result of intramolecular crosslinking of the subunits of a higher-order RT oligomer. A peptide that corresponds to part of the tryptophan repeat motif in the connection domain of HIV-1 RT inhibits crosslink formation as well as enzymatic activity. The crosslinking assay thus allows the investigation of the effect of inhibitors on the dimerization of HIV-1 RT. PMID:8745406

Auditory-motor interaction revealed by fMRI: speech, music, and working memory in area Spt.

PubMed

Hickok, Gregory; Buchsbaum, Bradley; Humphries, Colin; Muftuler, Tugan

2003-07-01

The concept of auditory-motor interaction pervades speech science research, yet the cortical systems supporting this interface have not been elucidated. Drawing on experimental designs used in recent work in sensory-motor integration in the cortical visual system, we used fMRI in an effort to identify human auditory regions with both sensory and motor response properties, analogous to single-unit responses in known visuomotor integration areas. The sensory phase of the task involved listening to speech (nonsense sentences) or music (novel piano melodies); the "motor" phase of the task involved covert rehearsal/humming of the auditory stimuli. A small set of areas in the superior temporal and temporal-parietal cortex responded both during the listening phase and the rehearsal/humming phase. A left lateralized region in the posterior Sylvian fissure at the parietal-temporal boundary, area Spt, showed particularly robust responses to both phases of the task. Frontal areas also showed combined auditory + rehearsal responsivity consistent with the claim that the posterior activations are part of a larger auditory-motor integration circuit. We hypothesize that this circuit plays an important role in speech development as part of the network that enables acoustic-phonetic input to guide the acquisition of language-specific articulatory-phonetic gestures; this circuit may play a role in analogous musical abilities. In the adult, this system continues to support aspects of speech production, and, we suggest, supports verbal working memory.

Kv channel subunits that contribute to voltage-gated K+ current in renal vascular smooth muscle.

PubMed

Fergus, Daniel J; Martens, Jeffrey R; England, Sarah K

2003-03-01

The rat renal arterial vasculature displays differences in K(+) channel current phenotypes along its length. Small arcuate to cortical radial arteries express a delayed rectifier phenotype, while the predominant Kv current in larger arcuate and interlobar arteries is composed of both transient and sustained components. We sought to determine whether Kvalpha subunits in the rat renal interlobar and arcuate arteries form heterotetramers, which may account for the unique currents, and whether modulatory Kvbeta subunits are present in renal vascular smooth muscle cells. RT-PCR indicated the presence of several different Kvalpha subunit isoform transcripts. Co-immunoprecipitation with immunoblotting and immunohistochemical evidence suggests that a portion of the K(+) current phenotype is a heteromultimer containing delayed-rectifier Kv1.2 and A-type Kv1.4 channel subunits. RT-PCR and immunoblot analyses also demonstrated the presence of both Kvbeta1.2 and Kvbeta1.3 in renal arteries. These results suggest that heteromultimeric formation of Kvalpha subunits and the presence of modulatory Kvbeta subunits are important factors in mediating Kv currents in the renal microvasculature and suggest a potentially critical role for these channel subunits in blood pressure regulation.

ADP binding to TF1 and its subunits induces ultraviolet spectral changes.

PubMed

Hisabori, T; Yoshida, M; Sakurai, H

1986-09-01

Adenine nucleotide binding sites on the coupling factor ATPase of thermophilic bacterium PS3 (TF1) were investigated by UV spectroscopy and by equilibrium dialysis. When ADP was mixed with TF1 in the presence and in the absence of Mg2+, an UV absorbance change was induced (t1/2 approximately 1 min) with a peak at about 278 nm and a trough at about 250 nm. Similar spectral changes were induced by ADP with the isolated beta subunits in the presence and in the absence of Mg2+, and with the isolated alpha subunits in the presence of Mg2+ although the magnitudes of the changes were different. From equilibrium dialysis measurement we identified two classes of nucleotide binding sites in TF1 in the presence of Mg2+, three high-affinity sites (Kd = 61 nM) and three low-affinity sites (Kd = 87 microM). In the absence of Mg2+, TF1 has one high-affinity site (Kd less than 10 nM) and five low-affinity sites (Kd = 100 microM). Moreover, we found a single Mg2+-dependent ADP binding site on the isolated alpha subunit and a single Mg2+-independent ADP binding site on the isolated beta subunit. From the above observations, we concluded that the three Mg2+-dependent high-affinity sites for ADP are located on the alpha subunit in TF1 and that the single high-affinity site is located on one of the beta subunits in TF1 in the absence of Mg2+.

β-Subunits of the SnRK1 Complexes Share a Common Ancestral Function Together with Expression and Function Specificities; Physical Interaction with Nitrate Reductase Specifically Occurs via AKINβ1-Subunit1[C][OA

PubMed Central

Polge, Cécile; Jossier, Mathieu; Crozet, Pierre; Gissot, Lionel; Thomas, Martine

2008-01-01

The SNF1/AMPK/SnRK1 kinases are evolutionary conserved kinases involved in yeast, mammals, and plants in the control of energy balance. These heterotrimeric enzymes are composed of one α-type catalytic subunit and two γ- and β-type regulatory subunits. In yeast it has been proposed that the β-type subunits regulate both the localization of the kinase complexes within the cell and the interaction of the kinases with their targets. In this work, we demonstrate that the three β-type subunits of Arabidopsis (Arabidopsis thaliana; AKINβ1, AKINβ2, and AKINβ3) restore the growth phenotype of the yeast sip1Δsip2Δgal83Δ triple mutant, thus suggesting the conservation of an ancestral function. Expression analyses, using AKINβ promoter∷β-glucuronidase transgenic lines, reveal different and specific patterns of expression for each subunit according to organs, developmental stages, and environmental conditions. Finally, our results show that the β-type subunits are involved in the specificity of interaction of the kinase with the cytosolic nitrate reductase. Together with previous cell-free phosphorylation data, they strongly support the proposal that nitrate reductase is a real target of SnRK1 in the physiological context. Altogether our data suggest the conservation of ancestral basic function(s) together with specialized functions for each β-type subunit in plants. PMID:18768910

Differences in the phenotypic effects of mutations in homologous MrpA and MrpD subunits of the multi-subunit Mrp-type Na+/H+ antiporter.

PubMed

Morino, Masato; Ogoda, Shinichiro; Krulwich, Terry Ann; Ito, Masahiro

2017-01-01

Mrp antiporters are the sole antiporters in the Cation/Proton Antiporter 3 family of transporter databases because of their unusual structural complexity, 6-7 hydrophobic proteins that function as a hetero-oligomeric complex. The two largest and homologous subunits, MrpA and MrpD, are essential for antiport activity and have direct roles in ion transport. They also show striking homology with proton-conducting, membrane-embedded Nuo subunits of respiratory chain complex I of bacteria, e.g., Escherichia coli. MrpA has the closest homology to the complex I NuoL subunit and MrpD has the closest homology to the complex I NuoM and N subunits. Here, introduction of mutations in MrpD, in residues that are also present in MrpA, led to defects in antiport function and/or complex formation. No significant phenotypes were detected in strains with mutations in corresponding residues of MrpA, but site-directed changes in the C-terminal region of MrpA had profound effects, showing that the MrpA C-terminal region has indispensable roles in antiport function. The results are consistent with a divergence in adaptations that support the roles of MrpA and MrpD in secondary antiport, as compared to later adaptations supporting homologs in primary proton pumping by the respiratory chain complex I.

Recent advances in the production of recombinant subunit vaccines in Pichia pastoris

PubMed Central

Wang, Man; Jiang, Shuai; Wang, Yefu

2016-01-01

ABSTRACT Recombinant protein subunit vaccines are formulated using defined protein antigens that can be produced in heterologous expression systems. The methylotrophic yeast Pichia pastoris has become an important host system for the production of recombinant subunit vaccines. Although many basic elements of P. pastoris expression system are now well developed, there is still room for further optimization of protein production. Codon bias, gene dosage, endoplasmic reticulum protein folding and culture condition are important considerations for improved production of recombinant vaccine antigens. Here we comment on current advances in the application of P. pastoris for the synthesis of recombinant subunit vaccines. PMID:27246656

5-Fluoroindole Resistance Identifies Tryptophan Synthase Beta Subunit Mutants in Arabidopsis Thaliana

PubMed Central

Barczak, A. J.; Zhao, J.; Pruitt, K. D.; Last, R. L.

1995-01-01

A study of the biochemical genetics of the Arabidopsis thaliana tryptophan synthase beta subunit was initiated by characterization of mutants resistant to the inhibitor 5-fluoroindole. Thirteen recessive mutations were recovered that are allelic to trp2-1, a mutation in the more highly expressed of duplicate tryptophan synthase beta subunit genes (TSB1). Ten of these mutations (trp2-2 through trp2-11) cause a tryptophan requirement (auxotrophs), whereas three (trp2-100 through trp2-102) remain tryptophan prototrophs. The mutations cause a variety of changes in tryptophan synthase beta expression. For example, two mutations (trp2-5 and trp2-8) cause dramatically reduced accumulation of TSB mRNA and immunologically detectable protein, whereas trp2-10 is associated with increased mRNA and protein. A correlation exists between the quantity of mutant beta and wild-type alpha subunit levels in the trp2 mutant plants, suggesting that the synthesis of these proteins is coordinated or that the quantity or structure of the beta subunit influences the stability of the alpha protein. The level of immunologically detectable anthranilate synthase alpha subunit protein is increased in the trp2 mutants, suggesting the possibility of regulation of anthranilate synthase levels in response to tryptophan limitation. PMID:7635295

Modulatory mechanisms and multiple functions of somatodendritic A-type K (+) channel auxiliary subunits.

PubMed

Jerng, Henry H; Pfaffinger, Paul J

2014-01-01

Auxiliary subunits are non-conducting, modulatory components of the multi-protein ion channel complexes that underlie normal neuronal signaling. They interact with the pore-forming α-subunits to modulate surface distribution, ion conductance, and channel gating properties. For the somatodendritic subthreshold A-type potassium (ISA) channel based on Kv4 α-subunits, two types of auxiliary subunits have been extensively studied: Kv channel-interacting proteins (KChIPs) and dipeptidyl peptidase-like proteins (DPLPs). KChIPs are cytoplasmic calcium-binding proteins that interact with intracellular portions of the Kv4 subunits, whereas DPLPs are type II transmembrane proteins that associate with the Kv4 channel core. Both KChIPs and DPLPs genes contain multiple start sites that are used by various neuronal populations to drive the differential expression of functionally distinct N-terminal variants. In turn, these N-terminal variants generate tremendous functional diversity across the nervous system. Here, we focus our review on (1) the molecular mechanism underlying the unique properties of different N-terminal variants, (2) the shaping of native ISA properties by the concerted actions of KChIPs and DPLP variants, and (3) the surprising ways that KChIPs and DPLPs coordinate the activity of multiple channels to fine-tune neuronal excitability. Unlocking the unique contributions of different auxiliary subunit N-terminal variants may provide an important opportunity to develop novel targeted therapeutics to treat numerous neurological disorders.

Facts First

DOE Office of Scientific and Technical Information (OSTI.GOV)

Wiley, H. S.

2009-02-01

I enjoy reading online news and opinions not only because of convenience, but also because of the responses from the readers. I know that these responses tend to be biased towards the extremes of opinions, but it is always interesting to see the diverse reactions from even seemingly non-controversial subjects. A recent example was the response to an editorial by Richard Gallaghar (October, 2008) that proffered the seemingly obvious opinion that science students need a strong understanding of scientific history and philosophy to help them face future challenges. This provoked a response from a reader, a college professor, that amore » major reason for the lack of good university science education was the laziness of his colleagues. They were taking the “easy way out” by concentrating on teaching facts rather than scientific concepts. In the respondent’s viewpoint, it was far better to teach students how to think. Facts could always be looked up in textbooks. If you did not understand how to think critically and create knowledge, you could not be considered educated. Although I am very sympathetic to this point of view, I don’t believe that laziness is the reason why students are flooded with facts rather than concepts in biology class.« less

Screening for AMPA receptor auxiliary subunit specific modulators

PubMed Central

Azumaya, Caleigh M.; Days, Emily L.; Vinson, Paige N.; Stauffer, Shaun; Sulikowski, Gary; Weaver, C. David; Nakagawa, Terunaga

2017-01-01

AMPA receptors (AMPAR) are ligand gated ion channels critical for synaptic transmission and plasticity. Their dysfunction is implicated in a variety of psychiatric and neurological diseases ranging from major depressive disorder to amyotrophic lateral sclerosis. Attempting to potentiate or depress AMPAR activity is an inherently difficult balancing act between effective treatments and debilitating side effects. A newly explored strategy to target subsets of AMPARs in the central nervous system is to identify compounds that affect specific AMPAR-auxiliary subunit complexes. This exploits diverse spatio-temporal expression patterns of known AMPAR auxiliary subunits, providing means for designing brain region-selective compounds. Here we report a high-throughput screening-based pipeline that can identify compounds that are selective for GluA2-CNIH3 and GluA2-stargazin complexes. These compounds will help us build upon the growing library of AMPAR-auxiliary subunit specific inhibitors, which have thus far all been targeted to TARP γ-8. We used a cell-based assay combined with a voltage-sensitive dye (VSD) to identify changes in glutamate-gated cation flow across the membranes of HEK cells co-expressing GluA2 and an auxiliary subunit. We then used a calcium flux assay to further validate hits picked from the VSD assay. VU0612951 and VU0627849 are candidate compounds from the initial screen that were identified as negative and positive allosteric modulators (NAM and PAM), respectively. They both have lower IC50/EC50s on complexes containing stargazin and CNIH3 than GSG1L or the AMPAR alone. We have also identified a candidate compound, VU0539491, that has NAM activity in GluA2(R)-CNIH3 and GluA2(Q) complexes and PAM activity in GluA2(Q)-GSG1L complexes. PMID:28358902

High affinity kainate receptor subunits are necessary for ionotropic but not metabotropic signaling

PubMed Central

Fernandes, Herman B.; Catches, Justin S.; Petralia, Ronald S.; Copits, Bryan A.; Xu, Jian; Russell, Theron A.; Swanson, Geoffrey T.; Contractor, Anis

2009-01-01

Summary Kainate receptors are atypical members of the glutamate receptor family which are able to signal through both ionotropic and metabotropic pathways. Of the five individual kainate receptor subunits the high-affinity subunits, GluK4 (KA1) and GluK5 (KA2), are unique in that they do not form functional homomeric receptors in recombinant expression systems, but combine with the primary subunits GluK1-3 (GluR5-7) to form heteromeric assemblies. Here we generated a GluK4 mutant mouse by disrupting the Grik4 gene locus. We found that loss of the GluK4 subunit leads to a significant reduction in synaptic kainate receptor currents. Moreover, ablation of both high-affinity subunits in GluK4/GluK5 double knockout mice leads to a complete loss of pre- and postsynaptic ionotropic function of synaptic kainate receptors. The principal subunits remain at the synaptic plasma membrane, but are distributed away from postsynaptic densities and presynaptic active zones. There is also an alteration in the properties of the remaining kainate receptors, as kainic acid application fails to elicit responses in GluK4/GluK5 knockout neurons. Despite the lack of detectable ionotropic synaptic receptors, the kainate receptor-mediated inhibition of the slow afterhyperpolarization current (IsAHP), which is dependent on metabotropic pathways, was intact in GluK4/GluK5 knockout mice. These results uncover a previously unknown critical role for the high-affinity kainate receptor subunits as obligatory components of ionotropic kainate receptor function, and further, demonstrate that kainate receptor participation in metabotropic signaling pathways does not require their classic role as ion channels. PMID:19778510

Use of a purified and functional recombinant calcium-channel beta4 subunit in surface-plasmon resonance studies.

PubMed Central

Geib, Sandrine; Sandoz, Guillaume; Mabrouk, Kamel; Matavel, Alessandra; Marchot, Pascale; Hoshi, Toshinori; Villaz, Michel; Ronjat, Michel; Miquelis, Raymond; Lévêque, Christian; de Waard, Michel

2002-01-01

Native high-voltage-gated calcium channels are multi-subunit complexes comprising a pore-forming subunit Ca(v) and at least two auxiliary subunits alpha(2)delta and beta. The beta subunit facilitates cell-surface expression of the channel and contributes significantly to its biophysical properties. In spite of its importance, detailed structural and functional studies are hampered by the limited availability of native beta subunit. Here, we report the purification of a recombinant calcium-channel beta(4) subunit from bacterial extracts by using a polyhistidine tag. The purified protein is fully functional since it binds on the alpha1 interaction domain, its main Ca(v)-binding site, and regulates the activity of P/Q calcium channel expressed in Xenopus oocytes in a similar way to the beta(4) subunit produced by cRNA injection. We took advantage of the functionality of the purified material to (i) develop an efficient surface-plasmon resonance assay of the interaction between two calcium channel subunits and (ii) measure, for the first time, the affinity of the recombinant His-beta(4) subunit for the full-length Ca(v)2.1 channel. The availability of this purified material and the development of a surface-plasmon resonance assay opens two immediate research perspectives: (i) drug screening programmes applied to the Ca(v)/beta interaction and (ii) crystallographic studies of the calcium-channel beta(4) subunit. PMID:11988102

Molecular evolution of Phox-related regulatory subunits for NADPH oxidase enzymes

PubMed Central

Kawahara, Tsukasa; Lambeth, J David

2007-01-01

Background The reactive oxygen-generating NADPH oxidases (Noxes) function in a variety of biological roles, and can be broadly classified into those that are regulated by subunit interactions and those that are regulated by calcium. The prototypical subunit-regulated Nox, Nox2, is the membrane-associated catalytic subunit of the phagocyte NADPH-oxidase. Nox2 forms a heterodimer with the integral membrane protein, p22phox, and this heterodimer binds to the regulatory subunits p47phox, p67phox, p40phox and the small GTPase Rac, triggering superoxide generation. Nox-organizer protein 1 (NOXO1) and Nox-activator 1 (NOXA1), respective homologs of p47phox and p67phox, together with p22phox and Rac, activate Nox1, a non-phagocytic homolog of Nox2. NOXO1 and p22phox also regulate Nox3, whereas Nox4 requires only p22phox. In this study, we have assembled and analyzed amino acid sequences of Nox regulatory subunit orthologs from vertebrates, a urochordate, an echinoderm, a mollusc, a cnidarian, a choanoflagellate, fungi and a slime mold amoeba to investigate the evolutionary history of these subunits. Results Ancestral p47phox, p67phox, and p22phox genes are broadly seen in the metazoa, except for the ecdysozoans. The choanoflagellate Monosiga brevicollis, the unicellular organism that is the closest relatives of multicellular animals, encodes early prototypes of p22phox, p47phox as well as the earliest known Nox2-like ancestor of the Nox1-3 subfamily. p67phox- and p47phox-like genes are seen in the sea urchin Strongylocentrotus purpuratus and the limpet Lottia gigantea that also possess Nox2-like co-orthologs of vertebrate Nox1-3. Duplication of primordial p47phox and p67phox genes occurred in vertebrates, with the duplicated branches evolving into NOXO1 and NOXA1. Analysis of characteristic domains of regulatory subunits suggests a novel view of the evolution of Nox: in fish, p40phox participated in regulating both Nox1 and Nox2, but after the appearance of mammals, Nox1

The morphology of cometary dust: Subunit size distributions down to tens of nanometres

NASA Astrophysics Data System (ADS)

Mannel, Thurid; Bentley, Mark; Boakes, Peter; Jeszenszky, Harald; Levasseur-Regourd, Anny-Chantal; Schmied, Roland; Torkar, Klaus

2017-04-01

The Rosetta orbiter carried a dedicated analysis suite for cometary dust. One of the key instruments was MIDAS (Micro-Imaging Dust Analysis System), an atomic force microscope that scanned the surfaces of hundreds of (sub-)micrometre particles in 3D with resolutions down to nanometres. This provided the opportunity to study the morphology of the smallest cometary dust; initial investigation revealed that the particles are agglomerates of smaller subunits [1] with different structural properties [2]. To understand the (surface-) structure of the dust particles and the origin of their smallest building blocks, a number of particles were investigated in detail and the size distribution of their subunits determined [3]. Here we discuss the subunit size distributions ranging from tens of nanometres to a few micrometres. The differences between the subunit size distributions for particles collected pre-perihelion, close to perihelion, and during a huge outburst are examined, as well as the dependence of subunit size on particle size. A case where a particle was fragmented in consecutive scans allows a direct comparison of fragment and subunit size distributions. Finally, the small end of the subunit size distribution is investigated: the smallest determined sizes will be reviewed in the context of other cometary missions, interplanetary dust particles believed to originate from comets, and remote observations. It will be discussed if the smallest subunits can be interpreted as fundamental building blocks of our early Solar System and if their origin was in our protoplanetary disc or the interstellar material. References: [1] M.S. Bentley, R. Schmied, T. Mannel et al., Aggregate dust particles at comet 67P/Chruyumov-Gerasimenko, Nature, 537, 2016. doi:10.1038/nature19091 [2] T. Mannel, M.S. Bentley, R. Schmied et al., Fractal cometary dust - a window into the early Solar system, MNRAS, 462, 2016. doi:10.1093/mnras/stw2898 [3] R. Schmied, T. Mannel, H. Jeszenszky, M

Activity-dependent control of NMDA receptor subunit composition at hippocampal mossy fibre synapses.

PubMed

Carta, Mario; Srikumar, Bettadapura N; Gorlewicz, Adam; Rebola, Nelson; Mulle, Christophe

2018-02-15

CA3 pyramidal cells display input-specific differences in the subunit composition of synaptic NMDA receptors (NMDARs). Although at low density, GluN2B contributes significantly to NMDAR-mediated EPSCs at mossy fibre synapses. Long-term potentiation (LTP) of NMDARs triggers a modification in the subunit composition of synaptic NMDARs by insertion of GluN2B. GluN2B subunits are essential for the expression of LTP of NMDARs at mossy fibre synapses. Single neurons express NMDA receptors (NMDARs) with distinct subunit composition and biophysical properties that can be segregated in an input-specific manner. The dynamic control of the heterogeneous distribution of synaptic NMDARs is crucial to control input-dependent synaptic integration and plasticity. In hippocampal CA3 pyramidal cells from mice of both sexes, we found that mossy fibre (MF) synapses display a markedly lower proportion of GluN2B-containing NMDARs than associative/commissural synapses. The mechanism involved in such heterogeneous distribution of GluN2B subunits is not known. Here we show that long-term potentiation (LTP) of NMDARs, which is selectively expressed at MF-CA3 pyramidal cell synapses, triggers a modification in the subunit composition of synaptic NMDARs by insertion of GluN2B. This activity-dependent recruitment of GluN2B at mature MF-CA3 pyramidal cell synapses contrasts with the removal of GluN2B subunits at other glutamatergic synapses during development and in response to activity. Furthermore, although expressed at low levels, GluN2B is necessary for the expression of LTP of NMDARs at MF-CA3 pyramidal cell synapses. Altogether, we reveal a previously unknown activity-dependent regulation and function of GluN2B subunits that may contribute to the heterogeneous plasticity induction rules in CA3 pyramidal cells. © 2017 Centre Nationnal de la Recherche Scientifique. The Journal of Physiology © 2017 The Physiological Society.

Fact Finders Go to Work!

ERIC Educational Resources Information Center

Gordon, Anitra

2005-01-01

Every class needs at least one certified Fact Finder. Most classes have at least one student who is years ahead of the class in reading ability and intellectual curiosity. The teacher and the library media specialist can identify one or more of these students in each class who might be delighted to become a certified Fact Finder. Fact Finders…

VAKT for Basic Subtraction Facts.

ERIC Educational Resources Information Center

Thornton, Carol A.; Toohey, Margaret A.

Guidelines are presented for modifying basic instruction of subtraction facts for elementary level learning disabled students. A detailed case study is used to illustrate a five-step structured program: (1) find a way to work it out; (2) add to check; (3) learn the partner facts; (4) study families of facts; (5) review and practice. The selection…

Structural basis of subunit selectivity for competitive NMDA receptor antagonists with preference for GluN2A over GluN2B subunits

DOE Office of Scientific and Technical Information (OSTI.GOV)

Lind, Genevieve E.; Mou, Tung-Chung; Tamborini, Lucia

NMDA-type glutamate receptors are ligand-gated ion channels that contribute to excitatory neurotransmission in the central nervous system (CNS). Most NMDA receptors comprise two glycine-binding GluN1 and two glutamate-binding GluN2 subunits (GluN2A–D). We describe highly potent (S)-5-[(R)-2-amino-2-carboxyethyl]-4,5-dihydro-1H-pyrazole-3-carboxylic acid (ACEPC) competitive GluN2 antagonists, of which ST3 has a binding affinity of 52 nM at GluN1/2A and 782 nM at GluN1/2B receptors. This 15-fold preference of ST3 for GluN1/2A over GluN1/2B is improved compared with NVP-AAM077, a widely used GluN2A-selective antagonist, which we show has 11-fold preference for GluN1/2A over GluN1/2B. Crystal structures of the GluN1/2A agonist binding domain (ABD) heterodimer with boundmore » ACEPC antagonists reveal a binding mode in which the ligands occupy a cavity that extends toward the subunit interface between GluN1 and GluN2A ABDs. Mutational analyses show that the GluN2A preference of ST3 is primarily mediated by four nonconserved residues that are not directly contacting the ligand, but positioned within 12 Å of the glutamate binding site. Two of these residues influence the cavity occupied by ST3 in a manner that results in favorable binding to GluN2A, but occludes binding to GluN2B. Thus, we reveal opportunities for the design of subunit-selective competitive NMDA receptor antagonists by identifying a cavity for ligand binding in which variations exist between GluN2A and GluN2B subunits. This structural insight suggests that subunit selectivity of glutamate-site antagonists can be mediated by mechanisms in addition to direct contributions of contact residues to binding affinity.« less

Structural basis of subunit selectivity for competitive NMDA receptor antagonists with preference for GluN2A over GluN2B subunits

PubMed Central

Lind, Genevieve E.; Mou, Tung-Chung; Tamborini, Lucia; Pomper, Martin G.; De Micheli, Carlo; Conti, Paola; Pinto, Andrea

2017-01-01

NMDA-type glutamate receptors are ligand-gated ion channels that contribute to excitatory neurotransmission in the central nervous system (CNS). Most NMDA receptors comprise two glycine-binding GluN1 and two glutamate-binding GluN2 subunits (GluN2A–D). We describe highly potent (S)-5-[(R)-2-amino-2-carboxyethyl]-4,5-dihydro-1H-pyrazole-3-carboxylic acid (ACEPC) competitive GluN2 antagonists, of which ST3 has a binding affinity of 52 nM at GluN1/2A and 782 nM at GluN1/2B receptors. This 15-fold preference of ST3 for GluN1/2A over GluN1/2B is improved compared with NVP-AAM077, a widely used GluN2A-selective antagonist, which we show has 11-fold preference for GluN1/2A over GluN1/2B. Crystal structures of the GluN1/2A agonist binding domain (ABD) heterodimer with bound ACEPC antagonists reveal a binding mode in which the ligands occupy a cavity that extends toward the subunit interface between GluN1 and GluN2A ABDs. Mutational analyses show that the GluN2A preference of ST3 is primarily mediated by four nonconserved residues that are not directly contacting the ligand, but positioned within 12 Å of the glutamate binding site. Two of these residues influence the cavity occupied by ST3 in a manner that results in favorable binding to GluN2A, but occludes binding to GluN2B. Thus, we reveal opportunities for the design of subunit-selective competitive NMDA receptor antagonists by identifying a cavity for ligand binding in which variations exist between GluN2A and GluN2B subunits. This structural insight suggests that subunit selectivity of glutamate-site antagonists can be mediated by mechanisms in addition to direct contributions of contact residues to binding affinity. PMID:28760974

Differential plasma membrane targeting of voltage-dependent calcium channel subunits expressed in a polarized epithelial cell line

PubMed Central

Brice, Nicola L; Dolphin, Annette C

1999-01-01

Voltage-dependent calcium channels (VDCCs) show a highly non-uniform distribution in many cell types, including neurons and other polarized secretory cells. We have examined whether this can be mimicked in a polarized epithelial cell line (Madin-Darby canine kidney), which has been used extensively to study the targeting of proteins. We expressed the VDCC α1A, α1B or α1C subunits either alone or in combination with accessory subunits α2-δ and the different β subunits, and examined their localization immunocytochemically. An α1 subunit was only targeted to the plasma membrane if co-expressed with the accessory subunits. The combination α1C/α2-δ and all β subunits was always localized predominantly to the basolateral membrane. It has been suggested that this is equivalent to somatodendritic targeting in neurons. In contrast, the α1B subunit was expressed at the apical membrane with all the accessory subunit combinations, by 24 h after microinjection. This membrane destination shows some parallels with axonal targeting in neurons. The α1A subunit was consistently observed at the apical membrane in the combinations α1A/α2-δ/β1b or β4. In contrast, when co-expressed with α2-δ/β2a, α1A was clearly targeted to the basolateral membrane. In conclusion, the VDCC α1 subunit appears to be the primary determinant for targeting the VDCC complex, but the β subunit can modify this destination, particularly for α1A. PMID:10066897

Loss of Drosophila Vps16A enhances autophagosome formation through reduced Tor activity.