Systematic analysis of viral genes responsible for differential virulence between American and Australian West Nile virus strains Free

Abstract

A variant Australian West Nile virus (WNV) strain, WNV, emerged in 2011 causing an unprecedented outbreak of encephalitis in horses in south-eastern Australia. However, no human cases associated with this strain have yet been reported. Studies using mouse models for WNV pathogenesis showed that WNV was less virulent than the human-pathogenic American strain of WNV, New York 99 (WNV). To identify viral genes and mutations responsible for the difference in virulence between WNV and WNV strains, we constructed chimeric viruses with substitution of large genomic regions coding for the structural genes, non-structural genes and untranslated regions, as well as seven individual non-structural gene chimeras, using a modified circular polymerase extension cloning method. Our results showed that the complete non-structural region of WNV, when substituted with that of WNV, significantly enhanced viral replication and the ability to suppress type I IFN response in cells, resulting in higher virulence in mice. Analysis of the individual non-structural gene chimeras showed a predominant contribution of WNV NS3 to increased virus replication and evasion of IFN response in cells, and to virulence in mice. Other WNV non-structural proteins (NS2A, NS4B and NS5) were shown to contribute to the modulation of IFN response. Thus a combination of non-structural proteins, likely NS2A, NS3, NS4B and NS5, is primarily responsible for the difference in virulence between WNV and WNV strains, and accumulative mutations within these proteins would likely be required for the Australian WNV strain to become significantly more virulent.

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2015-06-01
2024-03-29
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References

  1. Audsley M., Edmonds J., Liu W., Mokhonov V., Mokhonova E., Melian E. B., Prow N., Hall R. A., Khromykh A. A. 2011; Virulence determinants between New York 99 and Kunjin strains of West Nile virus. Virology 414:63–73 [View Article][PubMed]
    [Google Scholar]
  2. Beasley D. W., Li L., Suderman M. T., Barrett A. D. 2002; Mouse neuroinvasive phenotype of West Nile virus strains varies depending upon virus genotype. Virology 296:17–23 [View Article][PubMed]
    [Google Scholar]
  3. Beasley D. W., Davis C. T., Whiteman M., Granwehr B., Kinney R. M., Barrett A. D. 2004; Molecular determinants of virulence of West Nile virus in North America. Arch Virol Suppl 18:35–41[PubMed]
    [Google Scholar]
  4. Beasley D. W., Whiteman M. C., Zhang S., Huang C. Y., Schneider B. S., Smith D. R., Gromowski G. D., Higgs S., Kinney R. M., Barrett A. D. 2005; Envelope protein glycosylation status influences mouse neuroinvasion phenotype of genetic lineage 1 West Nile virus strains. J Virol 79:8339–8347 [View Article][PubMed]
    [Google Scholar]
  5. Borisevich V., Seregin A., Nistler R., Mutabazi D., Yamshchikov V. 2006; Biological properties of chimeric West Nile viruses. Virology 349:371–381 [View Article][PubMed]
    [Google Scholar]
  6. Brault A. C., Huang C. Y., Langevin S. A., Kinney R. M., Bowen R. A., Ramey W. N., Panella N. A., Holmes E. C., Powers A. M., Miller B. R. 2007; A single positively selected West Nile viral mutation confers increased virogenesis in American crows. Nat Genet 39:1162–1166 [View Article][PubMed]
    [Google Scholar]
  7. Cherng J. Y., Schuurmans-Nieuwenbroek N. M., Jiskoot W., Talsma H., Zuidam N. J., Hennink W. E., Crommelin D. J. 1999; Effect of DNA topology on the transfection efficiency of poly((2-dimethylamino)ethyl methacrylate)–plasmid complexes. J Control Release 60:343–353 [View Article][PubMed]
    [Google Scholar]
  8. Daffis S., Lazear H. M., Liu W. J., Audsley M., Engle M., Khromykh A. A., Diamond M. S. 2011; The naturally attenuated Kunjin strain of West Nile virus shows enhanced sensitivity to the host type I interferon response. J Virol 85:5664–5668 [View Article][PubMed]
    [Google Scholar]
  9. Ebel G. D., Fitzpatrick K. A., Lim P. Y., Bennett C. J., Deardorff E. R., Jerzak G. V., Kramer L. D., Zhou Y., Shi P. Y., Bernard K. A. 2011; Nonconsensus West Nile virus genomes arising during mosquito infection suppress pathogenesis and modulate virus fitness in vivo . J Virol 85:12605–12613 [View Article][PubMed]
    [Google Scholar]
  10. Edmonds J., van Grinsven E., Prow N., Bosco-Lauth A., Brault A. C., Bowen R. A., Hall R. A., Khromykh A. A. 2013; A novel bacterium-free method for generation of flavivirus infectious DNA by circular polymerase extension reaction allows accurate recapitulation of viral heterogeneity. J Virol 87:2367–2372 [View Article][PubMed]
    [Google Scholar]
  11. Frost M. J., Zhang J., Edmonds J. H., Prow N. A., Gu X., Davis R., Hornitzky C., Arzey K. E., Finlaison D. et al. 2012; Characterization of virulent West Nile virus Kunjin strain, Australia, 2011. Emerg Infect Dis 18:792–800 [View Article][PubMed]
    [Google Scholar]
  12. Hall R. A., Broom A. K., Smith D. W., Mackenzie J. S. 2002; The ecology and epidemiology of Kunjin virus. Curr Top Microbiol Immunol 267:253–269[PubMed]
    [Google Scholar]
  13. Henikoff S., Henikoff J. G. 1992; Amino acid substitution matrices from protein blocks. Proc Natl Acad Sci U S A 89:10915–10919 [View Article][PubMed]
    [Google Scholar]
  14. Hoenen A., Gillespie L., Morgan G., van der Heide P., Khromykh A., Mackenzie J. 2014; The West Nile virus assembly process evades the conserved antiviral mechanism of the interferon-induced MxA protein. Virology 448:104–116 [View Article][PubMed]
    [Google Scholar]
  15. Holzinger D., Jorns C., Stertz S., Boisson-Dupuis S., Thimme R., Weidmann M., Casanova J. L., Haller O., Kochs G. 2007; Induction of MxA gene expression by influenza A virus requires type I or type III interferon signaling. J Virol 81:7776–7785 [View Article][PubMed]
    [Google Scholar]
  16. Kajaste-Rudnitski A., Mashimo T., Frenkiel M. P., Guénet J. L., Lucas M., Desprès P. 2006; The 2′,5′-oligoadenylate synthetase 1b is a potent inhibitor of West Nile virus replication inside infected cells. J Biol Chem 281:4624–4637 [View Article][PubMed]
    [Google Scholar]
  17. Kim J. L., Morgenstern K. A., Griffith J. P., Dwyer M. D., Thomson J. A., Murcko M. A., Lin C., Caron P. R. 1998; Hepatitis C virus NS3 RNA helicase domain with a bound oligonucleotide: the crystal structure provides insights into the mode of unwinding. Structure 6:89–100 [View Article][PubMed]
    [Google Scholar]
  18. Lanciotti R. S., Ebel G. D., Deubel V., Kerst A. J., Murri S., Meyer R., Bowen M., McKinney N., Morrill W. E. et al. 2002; Complete genome sequences and phylogenetic analysis of West Nile virus strains isolated from the United States, Europe, and the Middle East. Virology 298:96–105 [View Article][PubMed]
    [Google Scholar]
  19. Laurent-Rolle M., Boer E. F., Lubick K. J., Wolfinbarger J. B., Carmody A. B., Rockx B., Liu W., Ashour J., Shupert W. L. et al. 2010; The NS5 protein of the virulent West Nile virus NY99 strain is a potent antagonist of type I interferon-mediated JAK-STAT signaling. J Virol 84:3503–3515 [View Article][PubMed]
    [Google Scholar]
  20. Mastrangelo E., Milani M., Bollati M., Selisko B., Peyrane F., Pandini V., Sorrentino G., Canard B., Konarev P. V. et al. 2007; Crystal structure and activity of Kunjin virus NS3 helicase; protease and helicase domain assembly in the full length NS3 protein. J Mol Biol 372:444–455 [View Article][PubMed]
    [Google Scholar]
  21. Mertens E., Kajaste-Rudnitski A., Torres S., Funk A., Frenkiel M. P., Iteman I., Khromykh A. A., Desprès P. 2010; Viral determinants in the NS3 helicase and 2K peptide that promote West Nile virus resistance to antiviral action of 2′,5′-oligoadenylate synthetase 1b. Virology 399:176–185 [View Article][PubMed]
    [Google Scholar]
  22. Patkar C. G., Kuhn R. J. 2008; Yellow fever virus NS3 plays an essential role in virus assembly independent of its known enzymatic functions. J Virol 82:3342–3352 [View Article][PubMed]
    [Google Scholar]
  23. Pletneva L. M., Haller O., Porter D. D., Prince G. A., Blanco J. C. 2008; Induction of type I interferons and interferon-inducible Mx genes during respiratory syncytial virus infection and reinfection in cotton rats. J Gen Virol 89:261–270 [View Article][PubMed]
    [Google Scholar]
  24. Prow N. A. 2013; The changing epidemiology of Kunjin virus in Australia. Int J Environ Res Public Health 10:6255–6272 [View Article][PubMed]
    [Google Scholar]
  25. Roby J., Funk A., Khromykh A. 2012; Flavivirus replication and assembly. In Molecular Virology and Control of Flaviviruses pp. 21–49 Edited by Shi P.-Y. Wymondham: Caister Academic Press;
    [Google Scholar]
  26. Roche S. E., Wicks R., Garner M. G., East I. J., Paskin R., Moloney B. J., Carr M., Kirkland P. 2013; Descriptive overview of the 2011 epidemic of arboviral disease in horses in Australia. Aust Vet J 91:5–13 [View Article][PubMed]
    [Google Scholar]
  27. Scherbik S. V., Kluetzman K., Perelygin A. A., Brinton M. A. 2007; Knock-in of the Oas1b(r) allele into a flavivirus-induced disease susceptible mouse generates the resistant phenotype. Virology 368:232–237 [View Article][PubMed]
    [Google Scholar]
  28. Simon-Chazottes D., Frenkiel M. P., Montagutelli X., Guénet J. L., Desprès P., Panthier J. J. 2011; Transgenic expression of full-length 2′,5′-oligoadenylate synthetase 1b confers to BALB/c mice resistance against West Nile virus-induced encephalitis. Virology 417:147–153 [View Article][PubMed]
    [Google Scholar]
  29. von Groll A., Levin Y., Barbosa M. C., Ravazzolo A. P. 2006; Linear DNA low efficiency transfection by liposome can be improved by the use of cationic lipid as charge neutralizer. Biotechnol Prog 22:1220–1224 [View Article][PubMed]
    [Google Scholar]
  30. Wu J., Bera A. K., Kuhn R. J., Smith J. L. 2005; Structure of the flavivirus helicase: implications for catalytic activity, protein interactions, and proteolytic processing. J Virol 79:10268–10277 [View Article][PubMed]
    [Google Scholar]
  31. Yamashita T., Unno H., Mori Y., Tani H., Moriishi K., Takamizawa A., Agoh M., Tsukihara T., Matsuura Y. 2008; Crystal structure of the catalytic domain of Japanese encephalitis virus NS3 helicase/nucleoside triphosphatase at a resolution of 1.8 Å. Virology 373:426–436 [View Article][PubMed]
    [Google Scholar]
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