Skip to main content
SpringerLink
Log in

Three amino acid changes in PB1-F2 of highly pathogenic H5N1 avian influenza virus affect pathogenicity in mallard ducks

  • Original Article
  • Published:
Archives of Virology Aims and scope Submit manuscript

Abstract

Despite reports that the PB1-F2 protein contributes to influenza virus pathogenicity in the mouse model, little is known about its significance in avian hosts. In our previous study, the A/Vietnam/1203/04 (H5N1) wild-type virus (wtVN1203) was more lethal to mallard ducks than a reverse genetics (rg)-derived VN1203. In search of potential viral factors responsible for this discrepancy, we found that synonymous mutations (SMs) had been inadvertently introduced into three genes of the rgVN1203 (rgVN1203/SM-3). Of 11 SMs in the PB1 gene, three resided in the PB1-F2 open reading frame, caused amino acid (aa) substitutions in the PB1-F2 protein, and reduced virus lethality in mallard ducks. The wtVN1203 and recombinant viruses with repairs to these three aa’s (rgVN1203/R-PB1-F2) or with repairs to all 11 SMs (rgVN1203/R-PB1) were significantly more pathogenic than rgVN1203/SM-3. In cultured cells, repairing three mutations in PB1-F2 increased viral polymerase activity and expression levels of viral RNA.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  1. Biswas SK, Nayak DP (1994) Mutational analysis of the conserved motifs of influenza A virus polymerase basic protein 1. J Virol 68:1819–1826

    CAS  PubMed  Google Scholar 

  2. Chen W, Calvo PA, Malide D, Gibbs J, Schubert U, Bacik I, Basta S, O’Neill R, Schickli J, Palese P, Henklein P, Bennink JR, Yewdell JW (2001) A novel influenza A virus mitochondrial protein that induces cell death. Nat Med 7:1306–1312

    Article  CAS  PubMed  Google Scholar 

  3. Coleman JR, Papamichail D, Skiena S, Futcher B, Wimmer E, Mueller S (2008) Virus attenuation by genome-scale changes in codon pair bias. Science 320:1784–1787

    Article  CAS  PubMed  Google Scholar 

  4. Conenello GM, Palese P (2007) Influenza A virus PB1-F2: a small protein with a big punch. Cell Host Microbe 2:207–209

    Article  CAS  PubMed  Google Scholar 

  5. Conenello GM, Zamarin D, Perrone LA, Tumpey T, Palese P (2007) A single mutation in the PB1-F2 of H5N1 (HK/97) and 1918 influenza A viruses contributes to increased virulence. PLoS Pathog 3:1414–1421

    Article  CAS  PubMed  Google Scholar 

  6. Ellis TM, Bousfield RB, Bissett LA, Dyrting KC, Luk GS, Tsim ST, Sturm-Ramirez K, Webster RG, Guan Y, Malik Peiris JS (2004) Investigation of outbreaks of highly pathogenic H5N1 avian influenza in waterfowl and wild birds in Hong Kong in late 2002. Avian Pathol 33:492–505

    Article  PubMed  Google Scholar 

  7. Faraway JJ (2006) Extending the Linear Model with R: generalized linear, mixed effects and nonparametric regression models. Chapman & Hall/CRC, Boca Raton, p 301

    Google Scholar 

  8. Gonzalez S, Zurcher T, Ortin J (1996) Identification of two separate domains in the influenza virus PB1 protein involved in the interaction with the PB2 and PA subunits: a model for the viral RNA polymerase structure. Nucleic Acids Res 24:4456–4463

    Article  CAS  PubMed  Google Scholar 

  9. Gonzalez S, Ortin J (1999) Distinct regions of influenza virus PB1 polymerase subunit recognize vRNA and cRNA templates. EMBO J 18:3767–3775

    Article  CAS  PubMed  Google Scholar 

  10. Hatta M, Gao P, Halfmann P, Kawaoka Y (2001) Molecular basis for high virulence of Hong Kong H5N1 influenza A viruses. Science 293:1840–1842

    Article  CAS  PubMed  Google Scholar 

  11. Hoffmann E, Neumann G, Hobom G, Webster RG, Kawaoka Y (2000) “Ambisense” approach for the generation of influenza A virus: vRNA and mRNA synthesis from one template. Virology 267:310–317

    Article  CAS  PubMed  Google Scholar 

  12. Hoffmann E, Neumann G, Kawaoka Y, Hobom G, Webster RG (2000) A DNA transfection system for generation of influenza A virus from eight plasmids. Proc Natl Acad Sci USA 97:6108–6113

    Article  CAS  PubMed  Google Scholar 

  13. Hoffmann E, Stech J, Guan Y, Webster RG, Perez DR (2001) Universal primer set for the full-length amplification of all influenza A viruses. Arch Virol 146:2275–2289

    Article  CAS  PubMed  Google Scholar 

  14. Hulse-Post DJ, Franks J, Boyd K, Salomon R, Hoffmann E, Yen HL, Webby RJ, Walker D, Nguyen TD, Webster RG (2007) Molecular changes in the polymerase genes (PA and PB1) associated with high pathogenicity of H5N1 influenza virus in mallard ducks. J Virol 81:8515–8524

    Article  CAS  PubMed  Google Scholar 

  15. Kerry PS, Willsher N, Fodor E (2008) A cluster of conserved basic amino acids near the C-terminus of the PB1 subunit of the influenza virus RNA polymerase is involved in the regulation of viral transcription. Virology 373:202–210

    Article  CAS  PubMed  Google Scholar 

  16. Kudla G, Murray AW, Tollervey D, Plotkin JB (2009) Coding-sequence determinants of gene expression in Escherichia coli. Science 324:255–258

    Article  CAS  PubMed  Google Scholar 

  17. Mazur I, Anhlan D, Mitzner D, Wixler L, Schubert U, Ludwig S (2008) The proapoptotic influenza A virus protein PB1-F2 regulates viral polymerase activity by interaction with the PB1 protein. Cell Microbiol 10:1140–1152

    Article  CAS  PubMed  Google Scholar 

  18. McAuley JL, Hornung F, Boyd KL, Smith AM, McKeon R, Bennink J, Yewdell JW, McCullers JA (2007) Expression of the 1918 influenza A virus PB1-F2 enhances the pathogenesis of viral and secondary bacterial pneumonia. Cell Host Microbe 2:240–249

    Article  CAS  PubMed  Google Scholar 

  19. McAuley JL, Zhang K, McCullers JA (2009) The effects of influenza A virus PB1-F2 protein on polymerase activity are strain specific and do not impact pathogenesis. J Virol

  20. Robb NC, Smith M, Vreede FT, Fodor E (2009) NS2/NEP protein regulates transcription and replication of the influenza virus RNA genome. J Gen Virol 90:1398–1407

    Article  CAS  PubMed  Google Scholar 

  21. Salomon R, Franks J, Govorkova EA, Ilyushina NA, Yen HL, Hulse-Post DJ, Humberd J, Trichet M, Rehg JE, Webby RJ, Webster RG, Hoffmann E (2006) The polymerase complex genes contribute to the high virulence of the human H5N1 influenza virus isolate A/Vietnam/1203/04. J Exp Med 203:689–697

    Article  CAS  PubMed  Google Scholar 

  22. Seo SH, Hoffmann E, Webster RG (2002) Lethal H5N1 influenza viruses escape host anti-viral cytokine responses. Nat Med 8:950–954

    Article  CAS  PubMed  Google Scholar 

  23. Sturm-Ramirez KM, Ellis T, Bousfield B, Bissett L, Dyrting K, Rehg JE, Poon L, Guan Y, Peiris M, Webster RG (2004) Reemerging H5N1 influenza viruses in Hong Kong in 2002 are highly pathogenic to ducks. J Virol 78:4892–4901

    Article  CAS  PubMed  Google Scholar 

  24. Torreira E, Schoehn G, Fernandez Y, Jorba N, Ruigrok RW, Cusack S, Ortin J, Llorca O (2007) Three-dimensional model for the isolated recombinant influenza virus polymerase heterotrimer. Nucleic Acids Res 35:3774–3783

    Article  CAS  PubMed  Google Scholar 

  25. Webster RG, Rott R (1987) Influenza virus A pathogenicity: the pivotal role of hemagglutinin. Cell 50:665–666

    Article  CAS  PubMed  Google Scholar 

  26. Yamada S, Suzuki Y, Suzuki T, Le MQ, Nidom CA, Sakai-Tagawa Y, Muramoto Y, Ito M, Kiso M, Horimoto T, Shinya K, Sawada T, Kiso M, Usui T, Murata T, Lin Y, Hay A, Haire LF, Stevens DJ, Russell RJ, Gamblin SJ, Skehel JJ, Kawaoka Y (2006) Haemagglutinin mutations responsible for the binding of H5N1 influenza A viruses to human-type receptors. Nature 444:378–382

    Article  CAS  PubMed  Google Scholar 

  27. Yen HL, Aldridge JR, Boon AC, Ilyushina NA, Salomon R, Hulse-Post DJ, Marjuki H, Franks J, Boltz DA, Bush D, Lipatov AS, Webby RJ, Rehg JE, Webster RG (2009) Changes in H5N1 influenza virus hemagglutinin receptor binding domain affect systemic spread. Proc Natl Acad Sci USA 106:286–291

    Article  CAS  PubMed  Google Scholar 

  28. Zamarin D, Ortigoza MB, Palese P (2006) Influenza A virus PB1-F2 protein contributes to viral pathogenesis in mice. J Virol 80:7976–7983

    Article  CAS  PubMed  Google Scholar 

  29. Zell R, Krumbholz A, Wutzler P (2006) Influenza A virus PB1-F2 gene. Emerg Infect Dis 12:1607–1608 author reply 1608–1609

    PubMed  Google Scholar 

Download references

Acknowledgments

This work was supported by the National Institutes of Health, U.S. Department of Health and Human Services, under Contract No. HHSN266200700005C, and by the American Lebanese Syrian Associated Charities (ALSAC). We thank the World Health Organization Global Influenza Surveillance Network for providing the parental A/VN/1203/04 (H5N1) virus. We acknowledge Richard Webby for advice and suggestions; Scott Krauss, Patrick Seiler, Heather Forrest, David Carey, and Cedric Proctor for excellent technical assistance; and James Knowles for administrative assistance. We greatly appreciate the gift of pARO3-vluci plasmid by Hui-Ling Yen and of pPolI A-luci plasmid by Stephan Pleschka. Scientific editing was provided by David Galloway and Sharon Naron.

Author information

Authors and Affiliations

  1. Division of Virology, Department of Infectious Diseases, St. Jude Children’s Research Hospital, Memphis, 262 Danny Thomas Place, Memphis, TN, 38105-3678, USA

    Henju Marjuki, Christoph Scholtissek, John Franks, Nicholas J. Negovetich, Jerry R. Aldridge, Rachelle Salomon & Robert G. Webster

  2. Hartwell Center for Bioinformatics and Biotechnology, St. Jude Children’s Research Hospital, Memphis, TN, 38105-3678, USA

    David Finkelstein

Authors
  1. Henju Marjuki
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Christoph Scholtissek
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. John Franks
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Nicholas J. Negovetich
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Jerry R. Aldridge
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Rachelle Salomon
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. David Finkelstein
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Robert G. Webster
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Robert G. Webster.

Additional information

H. Marjuki and C. Scholtissek contributed equally to this work.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Marjuki, H., Scholtissek, C., Franks, J. et al. Three amino acid changes in PB1-F2 of highly pathogenic H5N1 avian influenza virus affect pathogenicity in mallard ducks. Arch Virol 155, 925–934 (2010). https://doi.org/10.1007/s00705-010-0666-4

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00705-010-0666-4

Keywords

Search

Navigation