DOCSLIB.ORG

Mutation of Ebola Virus Matrix Protein Cysteine Residues Increases Binding to Phosphatidylserine Through Increased Flexibility of a Lipid Binding Loop

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Mutation of Ebola Virus Matrix Protein Cysteine Residues Increases Binding to Phosphatidylserine Through Increased Flexibility of a Lipid Binding Loop bioRxiv preprint doi: https://doi.org/10.1101/286120; this version posted March 21, 2018. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. Ebola virus VP40 Cys Mutations Increase PS Binding ! Mutation of Ebola Virus Matrix Protein Cysteine Residues Increases Binding to Phosphatidylserine through Increased Flexibility of a Lipid Binding Loop Kristen A. Johnson1, Nisha Bhattarai2, Melissa R. Budicini3, Carolyn M. Shirey1, Sarah Catherine B. Baker1, Bernard S. Gerstman2,4, Prem P. Chapagain2,4, and Robert V. Stahelin1,5,* 1 Department of Chemistry and Biochemistry, University of Notre Dame, Notre Dame, IN 46556, 2 Department of Physics, Florida International University, Miami, FL 33199, 3 Department of Biology, University of Notre Dame, Notre Dame, IN 46556, 4 Biomolecular Sciences Institute, Florida International University, Miami, FL 33199 5 Department of Medicinal Chemistry and Molecular Pharmacology Purdue University, West Lafayette, IN 47907. Running title: Ebola virus VP40 Cys Mutations Increase PS Binding *Address Correspondence to: Robert V. Stahelin, Purdue University, 575 Stadium Mall Drive, West Lafayette, IN 47907, TEL: 1-765-494-4152; FAX: 1-765-494-1414; Email: [email protected] Key Words: Ebola virus, matrix protein, membrane binding, phosphatidylserine, plasma membrane, viral budding, VP40 Abstract: promotes membrane binding loop flexibility, alters The Ebola virus (EBOV) is a genetically simple internal VP40 H-bonding, and increases PS negative sense RNA virus with only 7 genes yet it binding. To the best of our knowledge, this is the causes severe hemorrhagic fever in humans. The first evidence of mutations that increase VP40 matrix protein VP40 of EBOV is the main driver affinity for biological membranes and the length of viral budding through binding to host plasma of EBOV virus like particles. Together, our membrane lipids and formation of the filamentous, findings indicate these residues are important for pleomorphic virus particles. To better understand membrane dynamics at the plasma membrane via this dynamic and complex process we have asked the interaction with phosphatidylserine. what the role of two highly conserved cysteine residues are in the C-terminal domain of VP40. Introduction: Here we report that the mutation of Cys311 to The Ebola virus (EBOV) is a rare and alanine increases VP40 membrane binding affinity neglected disease that causes hemorrhagic fever in for phosphatidylserine containing membranes. humans with a high fatality rate. In 2014, the C311A has a significant increase in binding to PS largest EBOV outbreak in history began. The compared to WT, has longer virus like particles, outbreak became a global threat and over 3.6 and displays evidence of increased budding. billion dollars were spent to aid in containing the C314A also has an increase in PS binding outbreak and treating patients (1). This outbreak compared to WT, however to a lesser extent. The highlighted the importance of gaining a basic double Cys mutant shares the phenotypes of the understanding of how the EBOV components single mutants with increased binding to PS. function, as there are still no FDA approved Computational studies demonstrate these Cys therapies for treatment or prevention of infection. residues, Cys311 in particular, restrain a loop While vaccine and antibody therapy against the segment containing Lys residues that interact with EBOV glycoprotein has shown great promise, the plasma membrane. Mutation of Cys311 escape mutations against antibodies have been ! 1! bioRxiv preprint doi: https://doi.org/10.1101/286120; this version posted March 21, 2018. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. Ebola virus VP40 Cys Mutations Increase PS Binding ! detected as EBOV was passaged through animals cells, VP40 dependent budding is significantly (2) and a watch list of potential glycoprotein reduced (17,20,22) mutations that may evade antibody control has Cysteine is a rare and conserved amino been compiled (3). acid with only 2.26% occurrence in mammals and With only 8 proteins expressed from its higher eukaryotes, mathematically 0.865% less genome, EBOV is simple in design but is able to enriched than two codons should randomly perform many functions through the produce (26). Cysteine residues are important for transformative properties of the matrix protein, structural properties of proteins through disulfide VP40 (4). VP40 is essential for viral success as it bond formation and as a RNA, DNA, or histone is key for transcriptional regulation during binding protein through a zinc finger motif (27). infection (4), formation of the lipid-enveloped More than 21% of cysteine residues are found in a virus particles, and virus particle budding (5,6). C-(X)2-C motif, commonly attributed to metal Viral matrix protein flexibility is critical binding and oxidoreductase proteins (26). for multifunctional properties of VP40 and other Additionally, cysteine residues cannot engage in viral matrix proteins (7). Through flexible regions disulfide bond formation unless they have at least and different conformations, VP40 is able to bind two residues between them (26). RNA, lipids, and interact with an abundance of VP40 has two highly conserved cysteine host proteins to achieve transport to the plasma residues in the C-terminal domain of the protein. membrane and finally mediate viral budding (8- In many of the EBOV strains, these residues are 18). VP40 is sufficient form virus like particles found in a C-(X)2-C motif. In the crystal structure (VLPs) that bud from cells independent of the of VP40 from the Zaire type of EBOV (4LDB), other EBOV components (19). the cysteine residues are not engaged in a disulfide VP40 binds to anionic lipids at the plasma bond (4). This is not surprising as VP40 is membrane (PM) to form the host-derived lipid cytosolic and most cytosolic proteins do not have coat of the virus particle (20,21). VP40-lipid disulfide bonds as the reduced form is maintained binding is also essential for the formation of VP40 by thioredoxin proteins (26). Because cysteine hexamers (8,17,20) and the matrix sheet of large residues are low abundance residues, and VP40 oligomers (8,17) VP40 interacts with the plasma has two cysteine residues in a C-(X)2-C motif, we membrane through its C-terminal domain hypothesized that they play an important role in (4,18,20,22) and forms oligomers through NTD- VP40 function. NTD interactions (18). When VP40 binds to To test how these highly conserved anionic lipid phosphatidylserine (PS) hexamers are residues may impact VP40 function we examined formed in vitro and in live cells (17,23). Lipid the lipid-binding and oligomerization properties of induced VP40 oligomerization allows dissociation VP40 single or double cysteine to alanine of the N and C terminal domains that normally mutations in vitro and in cells. Additionally, we stabilize the protein in the closed dimer measured VLP formation, filament length, and conformation. Experimental data and molecular performed molecular dynamics simulations to dynamic simulations have shown that N- and C- investigate the molecular role of the Cys residues. terminal domain interactions, including several Here we show the cysteine residues play an salt-bridges, are important for maintaining the important role in regulation of PS binding and closed conformation of the protein (18,23,24). VP40 intramolecular interactions, loss of which VP40 plasma membrane binding and increases VP40 affinity for PS and increases the localization has been found to be mediated by length of filamentous virus like particles. phosphatidylserine (PS) and the low abundance inositol lipid, PI(4,5)P2. Both lipids are critical to Results: formation of VLPs where PS is important for initial plasma membrane recruitment and hexamer VP40 has two adjacent cysteine residues in the C- formation (17) and PI(4,5)P2 binding is essential terminal domain: for extensive VP40 oligomerization and oligomer VP40 cysteine residues are conserved in stability (25). When lipid binding is abolished in the Zaire, Tai Forrest, Bundibugyo, and Reston types but not the Sudan type of EBOV (Figure ! 2! bioRxiv preprint doi: https://doi.org/10.1101/286120; this version posted March 21, 2018. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. Ebola virus VP40 Cys Mutations Increase PS Binding ! 1A). The Sudan VP40 also has two cysteine mutations that increase the lipid-binding properties residues but they are located at slightly different of this viral matrix protein. positions, residues 314 and 320, respectively. Figure 1B shows the crystal structure (4LDB) VP40 cysteine residues have minor effects on with the two cysteine residues 311 (purple) and protein dynamics at the plasma membrane: 314 (blue) highlighted. These residues are located Because VP40 cysteine mutants have an within a solvent exposed, flexible loop on the side increase in binding to PS, an anionic lipid enriched of the C-terminal domain of the protein. The in the inner leaflet of the plasma membrane of host cysteine residues are not resolved in the Sudan cells that is critical for VP40 budding (17), we VP40 structure (4LD8) likely because they are expressed the EGFP-tagged constructs in live cells also located within a flexible loop region of the to observe the phenotype of the mutants. EGFP- protein. To investigate the role of these cysteine VP40-WT, EGFP-VP40-C311A, EGFP-VP40- residues in VP40 function, mutation of each Cys C314A, and EGFP-VP40-C311A/C314A were to Ala was made in 6xHis-VP40-pET46 and expressed in COS-7 cells. Cells expressing VLPs EGFP-VP40 for bacterial expression and protein or not were counted to see if the mutants had an purification and expression in cell culture for altered ability to form virus like particles.
Load more

Recommended publications
  • Identification of Hsc70 As an Influenza Virus Matrix Protein (M1)
    FEBS Letters 580 (2006) 5785–5790 Identification of Hsc70 as an influenza virus matrix protein (M1) binding factor involved in the virus life cycle Ken Watanabea, Takayuki Fusea, Issay Asanoa, Fujiko Tsukaharab, Yoshiro Marub, Kyosuke Nagatac, Kaio Kitazatoa, Nobuyuki Kobayashia,* a Laboratory of Molecular Biology of Infectious Agents, Graduate School of Biomedical Sciences, Nagasaki University, 1-14 Bunkyo-machi, Nagasaki 852-8521, Japan b Department of Pharmacology, Tokyo Women’s Medical University, School of Medicine, 8-1, Kawada-cho, Shinjuku-ku, Tokyo 162-8666, Japan c Institute of Basic Medical Sciences, University of Tsukuba, 1-1-1 Tennodai, Tsukuba 305-8575, Japan Received 22 August 2006; revised 13 September 2006; accepted 15 September 2006 Available online 27 September 2006 Edited by Felix Wieland nuclear export signal (NES), thus mechanism of M1-mediated Abstract Influenza virus matrix protein 1 (M1) has been shown to play a crucial role in the virus replication, assembly and bud- nuclear export of vRNP is still not well understood. ding. We identified heat shock cognate protein 70 (Hsc70) as a We herein identify heat shock cognate protein 70 (Hsc70), a M1 binding protein by immunoprecipitation and MALDI-TOF constitutive form of Hsp70 family protein, as an host factor(s) MS. The C terminal domain of M1 interacts with Hsc70. We which bind to M1 in infected cells by matrix-assisted laser found that Hsc70 does not correlate with the transport of M1 desorption ionization-time of flight mass spectrometry (MAL- to the nucleus, however, it does inhibit the nuclear export of DI-TOF MS). Heat shock proteins (Hsps) were induced by M1 and NP, thus resulting in the inhibition of viral production.
    [Show full text]
  • Distinct Contributions of Different Domains Within the HIV-1 Gag
    viruses Article Distinct Contributions of Different Domains within the HIV-1 Gag Polyprotein to Specific and Nonspecific Interactions with RNA Tomas Kroupa , Siddhartha A. K. Datta and Alan Rein * HIV Dynamics and Replication Program, Center for Cancer Research, National Cancer Institute, Frederick, MD 21702, USA; [email protected] (T.K.); [email protected] (S.A.K.D.) * Correspondence: [email protected] Received: 24 February 2020; Accepted: 31 March 2020; Published: 2 April 2020 Abstract: Viral genomic RNA is packaged into virions with high specificity and selectivity. However, in vitro the Gag specificity towards viral RNA is obscured when measured in buffers containing physiological salt. Interestingly, when the binding is challenged by increased salt concentration, the addition of competing RNAs, or introducing mutations to Gag protein, the specificity towards viral RNA becomes detectable. The objective of this work was to examine the contributions of the individual HIV-1 Gag polyprotein domains to nonspecific and specific RNA binding and stability of the initial protein-RNA complexes. Using a panel of Gag proteins with mutations disabling different Gag-Gag or Gag-RNA interfaces, we investigated the distinct contributions of individual domains which distinguish the binding to viral and nonviral RNA by measuring the binding of the proteins to RNAs. We measured the binding affinity in near-physiological salt concentration, and then challenged the binding by increasing the ionic strength to suppress the electrostatic interactions and reveal the contribution of specific Gag–RNA and Gag–Gag interactions. Surprisingly, we observed that Gag dimerization and the highly basic region in the matrix domain contribute significantly to the specificity of viral RNA binding.
    [Show full text]
  • How Influenza Virus Uses Host Cell Pathways During Uncoating
    cells Review How Influenza Virus Uses Host Cell Pathways during Uncoating Etori Aguiar Moreira 1 , Yohei Yamauchi 2 and Patrick Matthias 1,3,* 1 Friedrich Miescher Institute for Biomedical Research, 4058 Basel, Switzerland; [email protected] 2 Faculty of Life Sciences, School of Cellular and Molecular Medicine, University of Bristol, Bristol BS8 1TD, UK; [email protected] 3 Faculty of Sciences, University of Basel, 4031 Basel, Switzerland * Correspondence: [email protected] Abstract: Influenza is a zoonotic respiratory disease of major public health interest due to its pan- demic potential, and a threat to animals and the human population. The influenza A virus genome consists of eight single-stranded RNA segments sequestered within a protein capsid and a lipid bilayer envelope. During host cell entry, cellular cues contribute to viral conformational changes that promote critical events such as fusion with late endosomes, capsid uncoating and viral genome release into the cytosol. In this focused review, we concisely describe the virus infection cycle and highlight the recent findings of host cell pathways and cytosolic proteins that assist influenza uncoating during host cell entry. Keywords: influenza; capsid uncoating; HDAC6; ubiquitin; EPS8; TNPO1; pandemic; M1; virus– host interaction Citation: Moreira, E.A.; Yamauchi, Y.; Matthias, P. How Influenza Virus Uses Host Cell Pathways during 1. Introduction Uncoating. Cells 2021, 10, 1722. Viruses are microscopic parasites that, unable to self-replicate, subvert a host cell https://doi.org/10.3390/ for their replication and propagation. Despite their apparent simplicity, they can cause cells10071722 severe diseases and even pose pandemic threats [1–3].
    [Show full text]
  • Assessment of the Interaction Between the Human
    VARIATIONS IN THE V3 CROWN OF HIV-1 ENVELOPE IMPACT AFFINITY FOR CCR5 AND AFFECT ENTRY AND REPLICATIVE FITNESS By MICHAEL ANDREW LOBRITZ Submitted in partial fulfillment of the requirements for the degree of Doctor of Philosophy Dissertation advisor: Eric J. Arts, Ph.D. Department of Molecular Biology and Microbiology CASE WESTERN RESERVE UNIVERSITY August 2007 CASE WESTERN RESERVE UNIVERSITY SCHOOL OF GRADUATE STUDIES We hereby approve the dissertation of ______________________________________________________ candidate for the Ph.D. degree *. (signed)_______________________________________________ (chair of the committee) ________________________________________________ ________________________________________________ ________________________________________________ ________________________________________________ ________________________________________________ (date) _______________________ *We also certify that written approval has been obtained for any proprietary material contained therein. Table of Contents Chapter 1: Introduction........................................................................................................................16 1.A. HIV and AIDS..............................................................................................17 1.B. Retroviruses: Structure, Organization, and Replication...............................20 1.B.1. HIV-1 Genome...............................................................................20 1.B.2. HIV-1 Particle................................................................................23
    [Show full text]
  • Replication of HIV-1 Envelope Protein Cytoplasmic Domain Variants in Permissive and Restrictive Cells T
    Virology 538 (2019) 1–10 Contents lists available at ScienceDirect Virology journal homepage: www.elsevier.com/locate/virology Replication of HIV-1 envelope protein cytoplasmic domain variants in permissive and restrictive cells T ∗ August O. Staubus, Ayna Alfadhli, Robin Lid Barklis, Eric Barklis Department of Molecular Microbiology and Immunology, Oregon Health and Sciences University, 3181 SW Sam Jackson Park Road, Portland, OR, 97239, USA ARTICLE INFO ABSTRACT Keywords: Wild type (WT) HIV-1 envelope (Env) protein cytoplasmic tails (CTs) appear to be composed of membrane- HIV-1 proximal, N-terminal unstructured regions, and three C-terminal amphipathic helices. Previous studies have Replication shown that WT and CT-deleted (ΔCT) Env proteins are incorporated into virus particles via different mechan- Envelope protein isms. WT Env proteins traffic to cell plasma membranes (PMs), are rapidly internalized, recycle to PMs, and are Cytoplasmic tail incorporated into virions in permissive and restrictive cells in a Gag matrix (MA) protein-dependent fashion. In contrast, previously described ΔCT proteins do not appear to be internalized after their arrival to PMs, and do not require MA, but are only incorporated into virions in permissive cell lines. We have analyzed a new set of HIV-1 CT variants with respect to their replication in permissive and restrictive cells. Our results provide novel details as to how CT elements regulate HIV-1 Env protein function. 1. Introduction et al., 2018; Ohno et al., 1997; Wyss et al., 2001). Internalized Env proteins either are shunted off to lysosomes for degradation, or return The pathways by which HIV-1 envelope (Env) proteins become in- to PMs on recycling endosomes in a pathway that is dependent, in corporated into virions are complicated and incompletely elucidated certain cell types, on the Rab11 coupling protein FIP1C (Qi et al., 2013, (Checkley et al., 2011).
    [Show full text]
  • Phosphorylation of the M2 Protein of Influenza a Virus Is Not Essential for Virus Viability
    VIROLOGY 252, 54±64 (1998) ARTICLE NO. VY989384 Phosphorylation of the M2 Protein of Influenza A Virus Is Not Essential for Virus Viability Joanne M. Thomas, Mark P. Stevens, Neil Percy,1 and Wendy S. Barclay2 School of Animal and Microbial Sciences, University of Reading, Whiteknights, UK RG6 6AJ Received May 14, 1998; returned to author for revision June 12, 1998; accepted August 18, 1998 M2 is a minor component of the influenza A virus envelope. The cytoplasmic tail of the M2 protein is posttranslationally modified in the infected cell by palmitylation and phosphorylation. The primary site for phosphorylation of the M2 cytoplasmic tail is serine 64, which is highly conserved yet not required for the activity of the M2 ion channel. Using an exogenous incorporation assay, we have shown that incorporation of M2 into virus particles is type-specific and does not require phosphorylation of the cytoplasmic tail. In addition, phosphorylation of the cytoplasmic tail is not required for the directional transport of M2 in polarized MDCK cells. Using a reverse genetics and reassortment procedure, we generated a virus (Ra) specifically mutated in segment 7 such that the M2 cytoplasmic tail could no longer be phosphorylated. The virus was found to grow as well as wild-type virus in tissue culture and in eggs, was stable on passage in these systems, and possessed no second-site mutations in the engineered RNA segment. In vivo Ra replicated in Balb/c mice at least as well as the parent strain A/WSN/33. These studies indicate that phosphorylation of the M2 cytoplasmic tail is not required for in vitro or in vivo replication of influenza A virus.
    [Show full text]
  • Respiratory Syncytial Virus and Coronaviruses
    Chapter 3 Structural and Functional Aspects of Viroporins in Human Respiratory Viruses: Respiratory Syncytial Virus and Coronaviruses Wahyu Surya, Montserrat Samsó and Jaume Torres Additional information is available at the end of the chapter http://dx.doi.org/10.5772/53957 1. Introduction Viroporins are an increasingly recognized class of small viral membrane proteins (~60-120 amino acids) which oligomerize to produce hydrophilic pores at the membranes of virus- infected cells [1]. The existence of ‘viroporins’ was proposed more than 30 years ago after observing enhanced membrane permeability in infected cells [2]. These proteins form oligomers of defined size, and can act as proton or ion channels, and in general enhancing membrane permeability in the host [3]. Even though viroporins are not essential for the rep‐ lication of viruses, their absence results in attenuated or weakened viruses or changes in tropism (organ localization) and therefore diminished pathological effects [4, 5]. In addition to having one – sometimes two – α-helical transmembrane (TM) domain(s), viro‐ porins usually contain additional extramembrane regions that are able to make contacts with viral or host proteins. Indeed, the network of interactions of viroporins with other viral or cellular proteins is key to understand the regulation of viral protein trafficking through the vesicle system, viral morphogenesis and pathogenicity. In general, viroporins participate in the entry or release of viral particles into or out of cells, and membrane permeabilization may be a desirable functionality for the virus. Indeed, sev‐ eral viral proteins that are not viroporins are known to affect membrane permeabilization, e.g., A38L protein of vaccinia virus, a 33-kDa glycoprotein that allows Ca2+ influx and indu‐ ces necrosis in infected cells [6].
    [Show full text]
  • APICAL M2 PROTEIN IS REQUIRED for EFFICIENT INFLUENZA a VIRUS REPLICATION by Nicholas Wohlgemuth a Dissertation Submitted To
    APICAL M2 PROTEIN IS REQUIRED FOR EFFICIENT INFLUENZA A VIRUS REPLICATION by Nicholas Wohlgemuth A dissertation submitted to Johns Hopkins University in conformity with the requirements for the degree of Doctor of Philosophy Baltimore, Maryland October, 2017 © Nicholas Wohlgemuth 2017 All rights reserved ABSTRACT Influenza virus infections are a major public health burden around the world. This dissertation examines the influenza A virus M2 protein and how it can contribute to a better understanding of influenza virus biology and improve vaccination strategies. M2 is a member of the viroporin class of virus proteins characterized by their predicted ion channel activity. While traditionally studied only for their ion channel activities, viroporins frequently contain long cytoplasmic tails that play important roles in virus replication and disruption of cellular function. The currently licensed live, attenuated influenza vaccine (LAIV) contains a mutation in the M segment coding sequence of the backbone virus which confers a missense mutation (alanine to serine) in the M2 gene at amino acid position 86. Previously discounted for not showing a phenotype in immortalized cell lines, this mutation contributes to both the attenuation and temperature sensitivity phenotypes of LAIV in primary human nasal epithelial cells. Furthermore, viruses encoding serine at M2 position 86 induced greater IFN-λ responses at early times post infection. Reversing mutations such as this, and otherwise altering LAIV’s ability to replicate in vivo, could result in an improved LAIV development strategy. Influenza viruses infect at and egress from the apical plasma membrane of airway epithelial cells. Accordingly, the virus transmembrane proteins, HA, NA, and M2, are all targeted to the apical plasma membrane ii and contribute to egress.
    [Show full text]
  • Investigation of the C-Terminal Helix of HIV-1 Matrix: a Region Essential for Multiple Functions in the Viral Life Cycle: a Dissertation
    University of Massachusetts Medical School eScholarship@UMMS GSBS Dissertations and Theses Graduate School of Biomedical Sciences 2011-07-10 Investigation of the C-Terminal Helix of HIV-1 Matrix: A Region Essential for Multiple Functions in the Viral Life Cycle: A Dissertation Laura A. Brandano University of Massachusetts Medical School Let us know how access to this document benefits ou.y Follow this and additional works at: https://escholarship.umassmed.edu/gsbs_diss Part of the Biological Factors Commons, Genetic Phenomena Commons, Immunology and Infectious Disease Commons, Therapeutics Commons, Virology Commons, Virus Diseases Commons, and the Viruses Commons Repository Citation Brandano LA. (2011). Investigation of the C-Terminal Helix of HIV-1 Matrix: A Region Essential for Multiple Functions in the Viral Life Cycle: A Dissertation. GSBS Dissertations and Theses. https://doi.org/ 10.13028/5zmj-9x81. Retrieved from https://escholarship.umassmed.edu/gsbs_diss/552 This material is brought to you by eScholarship@UMMS. It has been accepted for inclusion in GSBS Dissertations and Theses by an authorized administrator of eScholarship@UMMS. For more information, please contact [email protected]. i INVESTIGATION OF THE C-TERMINAL HELIX OF HIV-1 MATRIX: A REGION ESSENTIAL FOR MULTIPLE FUNCTIONS IN THE VIRAL LIFE CYCLE A Dissertation Presented by Laura A. Brandano Submitted to the Faculty of the University of Massachusetts Graduate School of Biomedical Sciences, Worcester in partial fulfillment of the requirements for the degree of DOCTOR
    [Show full text]
  • XFEL Structures of the Influenza M2 Proton Channel: SPECIAL FEATURE Room Temperature Water Networks and Insights Into Proton Conduction
    XFEL structures of the influenza M2 proton channel: SPECIAL FEATURE Room temperature water networks and insights into proton conduction Jessica L. Thomastona, Rahel A. Woldeyesb, Takanori Nakane (中根 崇智)c, Ayumi Yamashitad, Tomoyuki Tanakad, Kotaro Koiwaie, Aaron S. Brewsterf, Benjamin A. Baradb, Yujie Cheng, Thomas Lemmina, Monarin Uervirojnangkoornh,i,j,k,l, Toshi Arimad, Jun Kobayashid, Tetsuya Masudad,m, Mamoru Suzukid,n, Michihiro Sugaharad, Nicholas K. Sauterf, Rie Tanakad, Osamu Nurekic, Kensuke Tonoo, Yasumasa Jotio, Eriko Nangod, So Iwatad,p, Fumiaki Yumotoe, James S. Fraserb, and William F. DeGradoa,1 aDepartment of Pharmaceutical Chemistry, University of California, San Francisco, CA 94158; bDepartment of Bioengineering and Therapeutic Sciences, University of California, San Francisco, CA 94158; cDepartment of Biological Sciences, Graduate School of Science, The University of Tokyo, Tokyo 113-0033, Japan; dSPring-8 Angstrom Compact Free Electron Laser (SACLA) Science Research Group, RIKEN SPring-8 Center, Saitama 351-0198, Japan; eStructural Biology Research Center, High Energy Accelerator Research Organization (KEK), Ibaraki 305-0801, Japan; fMolecular Biophysics and Integrated Bioimaging Division, Lawrence Berkeley National Laboratory, Berkeley, CA 94720; gSchool of Applied and Engineering Physics, Cornell University, Ithaca, NY 14853; hDepartment of Molecular and Cellular Physiology, Stanford University, Stanford, CA 94305; iHoward Hughes Medical Institute, Stanford University, Stanford, CA 94305; jDepartment of Neurology
    [Show full text]
  • Virus Entry, Assembly, Budding, and Membrane Rafts Nathalie Chazal, Denis Gerlier
    Virus Entry, Assembly, Budding, and Membrane Rafts Nathalie Chazal, Denis Gerlier To cite this version: Nathalie Chazal, Denis Gerlier. Virus Entry, Assembly, Budding, and Membrane Rafts. Microbi- ology and Molecular Biology Reviews, American Society for Microbiology, 2003, 67 (2), pp.226-237. 10.1128/MMBR.67.2.226-237.2003. hal-02147208 HAL Id: hal-02147208 https://hal.archives-ouvertes.fr/hal-02147208 Submitted on 7 Jun 2019 HAL is a multi-disciplinary open access L’archive ouverte pluridisciplinaire HAL, est archive for the deposit and dissemination of sci- destinée au dépôt et à la diffusion de documents entific research documents, whether they are pub- scientifiques de niveau recherche, publiés ou non, lished or not. The documents may come from émanant des établissements d’enseignement et de teaching and research institutions in France or recherche français ou étrangers, des laboratoires abroad, or from public or private research centers. publics ou privés. MICROBIOLOGY AND MOLECULAR BIOLOGY REVIEWS, June 2003, p. 226–237 Vol. 67, No. 2 1092-2172/03/$08.00ϩ0 DOI: 10.1128/MMBR.67.2.226–237.2003 Copyright © 2003, American Society for Microbiology. All Rights Reserved. Virus Entry, Assembly, Budding, and Membrane Rafts Nathalie Chazal1* and Denis Gerlier2 Immunologie-Virologie, EA 3038, Universite´Paul Sabatier, 31062 Toulouse,1 and Immunite´& Infections Virales, CNRS-UCBL UMR 5537, IFR Laennec, 69372 Lyon Cedex 08,2 France INTRODUCTION .......................................................................................................................................................226
    [Show full text]
  • Influenza Virus Matrix Protein M1 Preserves Its Conformation with Ph
    www.nature.com/scientificreports OPEN Infuenza virus Matrix Protein M1 preserves its conformation with pH, changing multimerization Received: 8 August 2017 Accepted: 21 November 2017 state at the priming stage due to Published: xx xx xxxx electrostatics Eleonora V. Shtykova1,2, Liubov A. Dadinova1, Natalia V. Fedorova3, Andrey E. Golanikov1, Elena N. Bogacheva2, Alexander L. Ksenofontov3, Liudmila A. Baratova3, Liudmila A. Shilova4,5, Vsevolod Yu. Tashkin4, Timur R. Galimzyanov4,6, Cy M. Jefries 7, Dmitri I. Svergun7 & Oleg V. Batishchev 4,5 Infuenza A virus matrix protein M1 plays an essential role in the virus lifecycle, but its functional and structural properties are not entirely defned. Here we employed small-angle X-ray scattering, atomic force microscopy and zeta-potential measurements to characterize the overall structure and association behavior of the full-length M1 at diferent pH conditions. We demonstrate that the protein consists of a globular N-terminal domain and a fexible C-terminal extension. The globular N-terminal domain of M1 monomers appears preserved in the range of pH from 4.0 to 6.8, while the C-terminal domain remains fexible and the tendency to form multimers changes dramatically. We found that the protein multimerization process is reversible, whereby the binding between M1 molecules starts to break around pH 6. A predicted electrostatic model of M1 self-assembly at diferent pH revealed a good agreement with zeta-potential measurements, allowing one to assess the role of M1 domains in M1-M1 and M1-lipid interactions. Together with the protein sequence analysis, these results provide insights into the mechanism of M1 scafold formation and the major role of the fexible and disordered C-terminal domain in this process.
    [Show full text]