STRUCTURE AND EXPRESSION OF THE PARAMYXOVIRUS SV5 GENOME

Summary

Principal Investigator: ROBERT LAMB
Abstract: The Paramyxoviridae are the etiological agents of many important diseases of man and lower animals. Collectively, these viruses contribute significantly to worldwide morbidity and mortality. The Paramyxoviridae include mumps virus, human parainfluenza virus type 1 (hPIV1), type 2, type 3, type 5 (also know as simian virus 5 [SV5]), Sendai virus, Newcastle disease virus, measles virus, canine distemper virus, rinderpest virus, respiratory syncytial virus, human metapneumovirus and the newly emergent highly pathogenic viruses, Hendra and Nipah viruses. The focus of this study is the mechanism by which paramyxoviruses enter cells. The entry of enveloped viruses into cells requires the fusion of the viral envelope with a cellular membrane. The mechanism of viral-mediated membrane fusion is a topic of interest to cell biologists and structural biologists as well as those investigating the mechanism of virus entry. This is because membrane fusion is a process central in cell biology. The class I viral fusion proteins are trapped in a meta-stable state and on activation these proteins undergo a significant re-folding event to achieve their final folded form and this protein re-folding drives membrane merger. Thus, studies on the mechanism by which paramyxoviruses cause cell fusion are of significance for understanding the pathogenesis of other class I viral fusion proteins such as gp120/gp41 of human immunodeficiency virus. For many paramyxoviruses two proteins on the virus surface, the hemagglutinin-neuraminidase (HN) and the fusion (F) protein are responsible for receptor binding and membrane fusion. This proposal is focused on providing biochemical data to support the notion that the F protein exists in a pre-fusion and a post-fusion state, to determine if the F protein cytoplasmic tail exists as a discrete protein structure that regulates fusion activity and to understand differences in the rate of virus-cell fusion and cell-cell fusion. We will elucidate the role of mutations in the F protein in the context of a virus infection by using reverse genetics and we will study fusion activity, virus growth and virus assembly of the altered viruses.
Funding Period: ----------------1986 - ---------------2011-
more information: NIH RePORT

Top Publications

  1. pmc Analysis of the pH requirement for membrane fusion of different isolates of the paramyxovirus parainfluenza virus 5
    Mei Lin Z Bissonnette
    Department of Biochemistry, Molecular Biology, and Cell Biology, Northwestern University, 2205 Tech Drive, Evanston, Illinois 60208 3500, USA
    J Virol 80:3071-7. 2006
  2. pmc Activation of paramyxovirus membrane fusion and virus entry
    Theodore S Jardetzky
    Department of Structural Biology, Stanford University School of Medicine, Stanford, CA 94305, United States Electronic address
    Curr Opin Virol 5:24-33. 2014
  3. pmc Fusion activation through attachment protein stalk domains indicates a conserved core mechanism of paramyxovirus entry into cells
    Sayantan Bose
    Department of Molecular Biosciences, Northwestern University, Evanston, Illinois, USA
    J Virol 88:3925-41. 2014
  4. pmc Mutations in the parainfluenza virus 5 fusion protein reveal domains important for fusion triggering and metastability
    Sayantan Bose
    Department of Molecular Biosciences
    J Virol 87:13520-31. 2013
  5. pmc Structure of the parainfluenza virus 5 (PIV5) hemagglutinin-neuraminidase (HN) ectodomain
    Brett D Welch
    Department of Molecular Biosciences, Northwestern University, Evanston, Illinois, USA
    PLoS Pathog 9:e1003534. 2013
  6. pmc Structure of the cleavage-activated prefusion form of the parainfluenza virus 5 fusion protein
    Brett D Welch
    Howard Hughes Medical Institute and Department of Molecular Biosciences, Northwestern University, Evanston, IL 60208, USA
    Proc Natl Acad Sci U S A 109:16672-7. 2012
  7. pmc Reversible inhibition of fusion activity of a paramyxovirus fusion protein by an engineered disulfide bond in the membrane-proximal external region
    Aarohi Zokarkar
    Department of Molecular Biosciences, Northwestern University, Evanston, Illinois, USA
    J Virol 86:12397-401. 2012
  8. pmc Fusion activation by a headless parainfluenza virus 5 hemagglutinin-neuraminidase stalk suggests a modular mechanism for triggering
    Sayantan Bose
    Department of Molecular Biosciences, Northwestern University, Evanston, IL 60208, USA
    Proc Natl Acad Sci U S A 109:E2625-34. 2012
  9. pmc Structure of the ulster strain newcastle disease virus hemagglutinin-neuraminidase reveals auto-inhibitory interactions associated with low virulence
    Ping Yuan
    Department of Structural Biology, Stanford University School of Medicine, Stanford, California, United States of America
    PLoS Pathog 8:e1002855. 2012
  10. ncbi Paramyxovirus membrane fusion: lessons from the F and HN atomic structures
    Robert A Lamb
    Howard Hughes Medical Institute, Northwestern University, Evanston, IL 60208 3500, USA
    Virology 344:30-7. 2006

Scientific Experts

  • Liqun Zhang
  • Robert A Lamb
  • Theodore S Jardetzky
  • George P Leser
  • Sayantan Bose
  • Ping Yuan
  • Sarah A Connolly
  • Brett D Welch
  • Aarohi Zokarkar
  • Reay G Paterson
  • Mei Lin Z Bissonnette
  • Christopher A Kors
  • Xiaolin Wen
  • Yong Ho Kim
  • Dana Ravid
  • Jason E Donald
  • Kurt Swanson
  • Jessica G Robach
  • William F Degrado
  • Benjamin J Chen
  • Marija Backovic
  • S Mark Tompkins
  • Albert S Song
  • Priya A Shah
  • Carissa M Heath
  • Maher Alayyoubi
  • John V Williams
  • James E Crowe
  • Jens C Krause
  • Reagan G Cox
  • Yuanyuan Liu
  • Gevorg Grigoryan
  • Kurt A Swanson
  • Alexander Y Fadeev
  • Borries Demeler
  • Richard Longnecker
  • Elizabeth W Howerth
  • Debra L Haas
  • Yuan Lin
  • Ralph Tripp
  • Kari A Kramer
  • Biao He
  • Joan E Durbin
  • Russell K Durbin
  • Jie Xu
  • Mary J Kennett
  • Hsien Shen Yin
  • Richard E Randall
  • Daniel F Young

Detail Information

Publications27

  1. pmc Analysis of the pH requirement for membrane fusion of different isolates of the paramyxovirus parainfluenza virus 5
    Mei Lin Z Bissonnette
    Department of Biochemistry, Molecular Biology, and Cell Biology, Northwestern University, 2205 Tech Drive, Evanston, Illinois 60208 3500, USA
    J Virol 80:3071-7. 2006
    ..The challenge of discovering how the paramyxovirus receptor binding protein (HN, H, or G) activates the metastable fusion protein to cause membrane fusion at neutral pH remains...
  2. pmc Activation of paramyxovirus membrane fusion and virus entry
    Theodore S Jardetzky
    Department of Structural Biology, Stanford University School of Medicine, Stanford, CA 94305, United States Electronic address
    Curr Opin Virol 5:24-33. 2014
    ....
  3. pmc Fusion activation through attachment protein stalk domains indicates a conserved core mechanism of paramyxovirus entry into cells
    Sayantan Bose
    Department of Molecular Biosciences, Northwestern University, Evanston, Illinois, USA
    J Virol 88:3925-41. 2014
    ....
  4. pmc Mutations in the parainfluenza virus 5 fusion protein reveal domains important for fusion triggering and metastability
    Sayantan Bose
    Department of Molecular Biosciences
    J Virol 87:13520-31. 2013
    ..The positions of the potential HN-interacting region and the region important for F stability in the lower part of the PIV5 F prefusion structure provide clues to the receptor-binding initiated, HN-mediated F trigger. ..
  5. pmc Structure of the parainfluenza virus 5 (PIV5) hemagglutinin-neuraminidase (HN) ectodomain
    Brett D Welch
    Department of Molecular Biosciences, Northwestern University, Evanston, Illinois, USA
    PLoS Pathog 9:e1003534. 2013
    ..The structure supports a model in which the heads of HN transition from down to up upon receptor binding thereby releasing steric constraints and facilitating the interaction between critical HN-stalk residues and F...
  6. pmc Structure of the cleavage-activated prefusion form of the parainfluenza virus 5 fusion protein
    Brett D Welch
    Howard Hughes Medical Institute and Department of Molecular Biosciences, Northwestern University, Evanston, IL 60208, USA
    Proc Natl Acad Sci U S A 109:16672-7. 2012
    ..The conformational freedom of the charged arginine residues that compose the protease recognition site increases on cleavage of F protein...
  7. pmc Reversible inhibition of fusion activity of a paramyxovirus fusion protein by an engineered disulfide bond in the membrane-proximal external region
    Aarohi Zokarkar
    Department of Molecular Biosciences, Northwestern University, Evanston, Illinois, USA
    J Virol 86:12397-401. 2012
    ..The data indicate that in addition to dissociation of the three-helix bundle stalk domain of prefusion F, the MPER region also needs to separate for F to be able to refold and cause fusion...
  8. pmc Fusion activation by a headless parainfluenza virus 5 hemagglutinin-neuraminidase stalk suggests a modular mechanism for triggering
    Sayantan Bose
    Department of Molecular Biosciences, Northwestern University, Evanston, IL 60208, USA
    Proc Natl Acad Sci U S A 109:E2625-34. 2012
    ....
  9. pmc Structure of the ulster strain newcastle disease virus hemagglutinin-neuraminidase reveals auto-inhibitory interactions associated with low virulence
    Ping Yuan
    Department of Structural Biology, Stanford University School of Medicine, Stanford, California, United States of America
    PLoS Pathog 8:e1002855. 2012
    ..These results clarify how the Ulster HN C-terminal residues lead to an auto-inhibited state of HN, the requirement for proteolytic activation of HNâ‚€ and associated reduced virulence...
  10. ncbi Paramyxovirus membrane fusion: lessons from the F and HN atomic structures
    Robert A Lamb
    Howard Hughes Medical Institute, Northwestern University, Evanston, IL 60208 3500, USA
    Virology 344:30-7. 2006
    ..In the last 5 years, atomic structures of paramyxovirus F and HN proteins have been reported. The knowledge gained from these structures towards understanding the mechanism of viral membrane fusion is described...
  11. ncbi Paramyxovirus fusion: real-time measurement of parainfluenza virus 5 virus-cell fusion
    Sarah A Connolly
    Howard Hughes Medical Institute, Northwestern University, Evanston, IL 60208 3500, USA
    Virology 355:203-12. 2006
    ..These results support the notion that virus-cell and cell-cell fusion have significant differences...
  12. pmc Refolding of a paramyxovirus F protein from prefusion to postfusion conformations observed by liposome binding and electron microscopy
    Sarah A Connolly
    Howard Hughes Medical Institute, Northwestern University, Evanston, IL 60208 3500, USA
    Proc Natl Acad Sci U S A 103:17903-8. 2006
    ..The reactivity of the F protein with conformation-specific mAbs and peptide binding suggest that soluble F-GCNt and membrane-bound F proteins refold through a comparable pathway...
  13. pmc Recombinant parainfluenza virus 5 (PIV5) expressing the influenza A virus hemagglutinin provides immunity in mice to influenza A virus challenge
    S Mark Tompkins
    Department of Infectious Diseases, University of Georgia, Athens, GA, USA
    Virology 362:139-50. 2007
    ..The efficacy of rPIV5-H3 as a live vaccine was examined in 6-week-old BALB/c mice. The results show that a single dose inoculation provides broad and considerable immunity against influenza A virus infection...
  14. pmc Characterization of EBV gB indicates properties of both class I and class II viral fusion proteins
    Marija Backovic
    Department of Biochemistry, Molecular Biology, Cell Biology, Northwestern University, Evanston, IL 60208, USA
    Virology 368:102-13. 2007
    ..These data demonstrate biochemical features of EBV gB that are characteristic of other class I and class II viral fusion proteins, but not of HSV-1 gB...
  15. pmc Mechanisms for enveloped virus budding: can some viruses do without an ESCRT?
    Benjamin J Chen
    Department of Biochemistry, Molecular Biology and Cell Biology, Northwestern University, Evanston, Illinois 60208 3500, USA
    Virology 372:221-32. 2008
    ....
  16. pmc Domain architecture and oligomerization properties of the paramyxovirus PIV 5 hemagglutinin-neuraminidase (HN) protein
    Ping Yuan
    Department of Structural Biology, Stanford University, Palo Alto, CA 94305 5126, USA
    Virology 378:282-91. 2008
    ..Electron microscopy of the HN ectodomain reveals flexible arrangements of the NA and stalk domains, which may be important for understanding how these two HN domains impact virus entry...
  17. pmc Structure of the human metapneumovirus fusion protein with neutralizing antibody identifies a pneumovirus antigenic site
    Xiaolin Wen
    Department of Structural Biology, Stanford University School of Medicine, Stanford, California, USA
    Nat Struct Mol Biol 19:461-3. 2012
    ..Here we show that a potently neutralizing anti-human metapneumovirus antibody (DS7) binds a structurally invariant domain of F, revealing a new epitope that could be targeted in vaccine development...
  18. pmc Functional analysis of the transmembrane domain in paramyxovirus F protein-mediated membrane fusion
    Mei Lin Z Bissonnette
    Department of Biochemistry, Molecular Biology and Cell Biology, Northwestern University, Evanston, IL 60208 3500, USA
    J Mol Biol 386:14-36. 2009
    ..We suggest that the unusual length of this TM helix might allow it to serve as a template for formation of or specifically stabilize the lipid stalk intermediate in fusion...
  19. pmc Bimolecular complementation of paramyxovirus fusion and hemagglutinin-neuraminidase proteins enhances fusion: implications for the mechanism of fusion triggering
    Sarah A Connolly
    Howard Hughes Medical Institute
    J Virol 83:10857-68. 2009
    ..In support of a provocateur model of F activation, we demonstrate by analysis of the morphology of soluble F trimers that the hyperfusogenic mutation S443P has a destabilizing effect on F...
  20. pmc Capture and imaging of a prehairpin fusion intermediate of the paramyxovirus PIV5
    Yong Ho Kim
    Department of Chemistry, University of Pennsylvania, Philadelphia, PA 19104, USA
    Proc Natl Acad Sci U S A 108:20992-7. 2011
    ....
  21. pmc The paramyxovirus fusion protein C-terminal region: mutagenesis indicates an indivisible protein unit
    Aarohi Zokarkar
    Department of Molecular Biosciences, Northwestern University, Evanston, Illinois, USA
    J Virol 86:2600-9. 2012
    ..Alanine scanning mutagenesis of MPER indicates that it has a regulatory role in fusion since both hyperfusogenic and hypofusogenic mutations were found...
  22. pmc Structure and mutagenesis of the parainfluenza virus 5 hemagglutinin-neuraminidase stalk domain reveals a four-helix bundle and the role of the stalk in fusion promotion
    Sayantan Bose
    Department of Molecular Biosciences, Northwestern University, 2205 Tech Drive, Evanston, IL 60208 3500, USA
    J Virol 85:12855-66. 2011
    ....
  23. pmc Comparison of differing cytopathic effects in human airway epithelium of parainfluenza virus 5 (W3A), parainfluenza virus type 3, and respiratory syncytial virus
    Liqun Zhang
    Cystic Fibrosis Pulmonary Research and Treatment Center, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina 27599, USA
    Virology 421:67-77. 2011
    ..These studies revealed striking differences in cytopathology of PIV5 versus PIV3 or RSV and indicate the extent of cytopathology determined in cell-lines does not predict events in differentiated airway cells...
  24. pmc Structure of the Newcastle disease virus hemagglutinin-neuraminidase (HN) ectodomain reveals a four-helix bundle stalk
    Ping Yuan
    Department of Structural Biology, Stanford University School of Medicine, Stanford, CA 94305, USA
    Proc Natl Acad Sci U S A 108:14920-5. 2011
    ..Two of four NA domains interact with the 4HB stalk, and residues at this interface in both the stalk and NA domain have been implicated in HN function...
  25. pmc Analysis of parainfluenza virus-5 hemagglutinin-neuraminidase protein mutants that are blocked in internalization and degradation
    Jessica G Robach
    Department of Biochemistry, Molecular Biology, and Cell Biology, Northwestern University, Evanston, IL 60208 3500, USA
    Virology 406:189-201. 2010
    ..Furthermore, oligomeric analyses indicate that HN E37 mutants perturb the tetrameric organization of HN, probably by destabilizing the dimer-of-dimers interface...
  26. pmc A role for caveolin 1 in assembly and budding of the paramyxovirus parainfluenza virus 5
    Dana Ravid
    Howard Hughes Medical Institute, Northwestern University, Evanston, IL 60208 3500, USA
    J Virol 84:9749-59. 2010
    ..These data suggest that Cav-1 affects assembly and/or budding, and this is supported by the finding that Cav-1 is incorporated into mature viral particles...
  27. pmc Structure of the Newcastle disease virus F protein in the post-fusion conformation
    Kurt Swanson
    Howard Hughes Medical Institute, USA
    Virology 402:372-9. 2010
    ..Electrostatic and temperature factor analysis of the F structures points to regions of these proteins that may be functionally important in their membrane fusion activity...