1887

Abstract

Human metapneumovirus (HMPV) is an important cause of respiratory tract infections. The mechanism by which its fusion (F) protein is responsible for variable cytopathic effects remains unknown. We aligned the F sequences of the poorly fusogenic B2/CAN98-75 strain and the hyperfusogenic A1/C-85473 strain and identified divergent residues located in the two functional heptad repeats domains (HRA and HRB). We generated recombinant viruses by inserting the mutations N135T–G139N–T143K–K166E–E167D in HRA and/or K479R–N482S in HRB, corresponding to swapped sequences from C-85473, into CAN98-75 background and investigated their impact on phenotype and fusogenicity. We demonstrated that the five HRA mutations enhanced the fusogenicity of the recombinant rCAN98-75 virus, almost restoring the phenotype of the wild-type rC-85473 strain, whereas HRB substitutions alone had no significant effect on cell–cell fusion. Altogether, our results support the importance of the HRA domain for an HMPV-triggered fusion mechanism and identify key residues that modulate syncytium formation.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/jgv.0.000796
2017-06-01
2024-03-28
Loading full text...

Full text loading...

/deliver/fulltext/jgv/98/6/1174.html?itemId=/content/journal/jgv/10.1099/jgv.0.000796&mimeType=html&fmt=ahah

References

  1. Panda S, Mohakud NK, Pena L, Kumar S. Human metapneumovirus: review of an important respiratory pathogen. Int J Infect Dis 2014; 25:45–52 [View Article][PubMed]
    [Google Scholar]
  2. van den Hoogen BG, Bestebroer TM, Osterhaus AD, Fouchier RA. Analysis of the genomic sequence of a human metapneumovirus. Virology 2002; 295:119–132 [View Article][PubMed]
    [Google Scholar]
  3. Peret TC, Boivin G, Li Y, Couillard M, Humphrey C et al. Characterization of human metapneumoviruses isolated from patients in North America. J Infect Dis 2002; 185:1660–1663 [View Article][PubMed]
    [Google Scholar]
  4. Biacchesi S, Skiadopoulos MH, Boivin G, Hanson CT, Murphy BR et al. Genetic diversity between human metapneumovirus subgroups. Virology 2003; 315:1–9 [View Article][PubMed]
    [Google Scholar]
  5. van den Hoogen BG, Herfst S, Sprong L, Cane PA, Forleo-Neto E et al. Antigenic and genetic variability of human metapneumoviruses. Emerg Infect Dis 2004; 10:658–666 [View Article][PubMed]
    [Google Scholar]
  6. ICTV ICoToV 2015; Elevation of the paramyxoviral subfamily Pneumovirinae to family status as family Pneumovirinae in the order mononegavirales; and renaming of one pneumoviral genus. https://talk.ictvonline.org/ICTV/proposals/2015.011a-iM.A.v2.Pneumoviridae.pdf
  7. Hamelin ME, Abed Y, Boivin G. Human metapneumovirus: a new player among respiratory viruses. Clin Infect Dis 2004; 38:983–990 [View Article][PubMed]
    [Google Scholar]
  8. Papenburg J, Boivin G. The distinguishing features of human metapneumovirus and respiratory syncytial virus. Rev Med Virol 2010; 20:245–260 [View Article][PubMed]
    [Google Scholar]
  9. Feuillet F, Lina B, Rosa-Calatrava M, Boivin G. Ten years of human metapneumovirus research. J Clin Virol 2012; 53:97–105 [View Article][PubMed]
    [Google Scholar]
  10. Le Bayon JC, Lina B, Rosa-Calatrava M, Boivin G. Recent developments with live-attenuated recombinant paramyxovirus vaccines. Rev Med Virol 2013; 23:15–34 [View Article][PubMed]
    [Google Scholar]
  11. Chang A, Dutch RE. Paramyxovirus fusion and entry: multiple paths to a common end. Viruses 2012; 4:613–636 [View Article][PubMed]
    [Google Scholar]
  12. Cox RG, Williams JV. Breaking in: human metapneumovirus fusion and entry. Viruses 2013; 5:192–210 [View Article][PubMed]
    [Google Scholar]
  13. Schowalter RM, Smith SE, Dutch RE. Characterization of human metapneumovirus F protein-promoted membrane fusion: critical roles for proteolytic processing and low pH. J Virol 2006; 80:10931–10941 [View Article][PubMed]
    [Google Scholar]
  14. Lamb RA, Jardetzky TS. Structural basis of viral invasion: lessons from paramyxovirus F. Curr Opin Struct Biol 2007; 17:427–436 [View Article][PubMed]
    [Google Scholar]
  15. Harrison SC. Viral membrane fusion. Virology 2015; 479-480:498–507 [View Article][PubMed]
    [Google Scholar]
  16. Aerts L, Cavanagh MH, Dubois J, Carbonneau J, Rhéaume C et al. Effect of in vitro syncytium formation on the severity of human metapneumovirus disease in a murine model. PLoS One 2015; 10:e0120283 [View Article][PubMed]
    [Google Scholar]
  17. Luque LE, Russell CJ. Spring-loaded heptad repeat residues regulate the expression and activation of paramyxovirus fusion protein. J Virol 2007; 81:3130–3141 [View Article][PubMed]
    [Google Scholar]
  18. Chang A, Hackett BA, Winter CC, Buchholz UJ, Dutch RE. Potential electrostatic interactions in multiple regions affect human metapneumovirus F-mediated membrane fusion. J Virol 2012; 86:9843–9853 [View Article][PubMed]
    [Google Scholar]
  19. Terrier O, Durupt F, Cartet G, Thomas L, Lina B et al. Engineering of a parainfluenza virus type 5 fusion protein (PIV-5 F): development of an autonomous and hyperfusogenic protein by a combinational mutagenesis approach. Virus Res 2009; 146:115–124 [View Article][PubMed]
    [Google Scholar]
  20. Mas V, Herfst S, Osterhaus AD, Fouchier RA, Melero JA. Residues of the human metapneumovirus fusion (F) protein critical for its strain-related fusion phenotype: implications for the virus replication cycle. J Virol 2011; 85:12650–12661 [View Article][PubMed]
    [Google Scholar]
  21. Herfst S, Mas V, ver LS, Wierda RJ, Osterhaus AD et al. Low-pH-induced membrane fusion mediated by human metapneumovirus F protein is a rare, strain-dependent phenomenon. J Virol 2008; 82:8891–8895 [View Article][PubMed]
    [Google Scholar]
  22. Schowalter RM, Chang A, Robach JG, Buchholz UJ, Dutch RE. Low-pH triggering of human metapneumovirus fusion: essential residues and importance in entry. J Virol 2009; 83:1511–1522 [View Article][PubMed]
    [Google Scholar]
  23. Boivin G, Mackay I, Sloots TP, Madhi S, Freymuth F et al. Global genetic diversity of human metapneumovirus fusion gene. Emerg Infect Dis 2004; 10:1154–1157 [View Article][PubMed]
    [Google Scholar]
  24. Deffrasnes C, Hamelin ME, Prince GA, Boivin G. Identification and evaluation of a highly effective fusion inhibitor for human metapneumovirus. Antimicrob Agents Chemother 2008; 52:279–287 [View Article][PubMed]
    [Google Scholar]
  25. Papenburg J, Carbonneau J, Isabel S, Bergeron MG, Williams JV et al. Genetic diversity and molecular evolution of the major human metapneumovirus surface glycoproteins over a decade. J Clin Virol 2013; 58:541–547 [View Article][PubMed]
    [Google Scholar]
  26. Meng J, Hotard AL, Currier MG, Lee S, Stobart CC et al. Respiratory syncytial virus attachment glycoprotein contribution to infection depends on the specific fusion protein. J Virol 2015; 90:245–253 [View Article][PubMed]
    [Google Scholar]
  27. Gardner AE, Dutch RE. A conserved region in the F(2) subunit of paramyxovirus fusion proteins is involved in fusion regulation. J Virol 2007; 81:8303–8314 [View Article][PubMed]
    [Google Scholar]
  28. Biasini M, Bienert S, Waterhouse A, Arnold K, Studer G et al. SWISS-MODEL: modelling protein tertiary and quaternary structure using evolutionary information. Nucleic Acids Res 2014; 42:W252–W258 [View Article][PubMed]
    [Google Scholar]
  29. Más V, Rodriguez L, Olmedillas E, Cano O, Palomo C et al. Engineering, structure and immunogenicity of the human metapneumovirus F protein in the postfusion conformation. PLoS Pathog 2016; 12:e1005859 [View Article][PubMed]
    [Google Scholar]
  30. Lawlor HA, Schickli JH, Tang RS. A single amino acid in the F2 subunit of respiratory syncytial virus fusion protein alters growth and fusogenicity. J Gen Virol 2013; 94:2627–2635 [View Article][PubMed]
    [Google Scholar]
  31. Le Bayon JC, Terrier O, Cartet G, Lina B, Rosa-Calatrava M. Characterization of a key residue for hyperfusogenic phenotype in human parainfluenza virus type 2 (hPIV-2) fusion glycoprotein. Virus Genes 2013; 47:365–369 [View Article][PubMed]
    [Google Scholar]
  32. Singh M, Berger B, Kim PS. LearnCoil-VMF: computational evidence for coiled–coil-like motifs in many viral membrane fusion proteins. J Mol Biol 1999; 290:1031–1041 [View Article][PubMed]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/jgv.0.000796
Loading
/content/journal/jgv/10.1099/jgv.0.000796
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error