Influence of PB2 host-range determinants on the intranuclear mobility of the influenza A virus polymerase Open Access

Abstract

Avian influenza A viruses often do not propagate efficiently in mammalian cells. The viral polymerase protein PB2 is important for this host restriction, with amino-acid polymorphisms at residue 627 and other positions acting as ‘signatures’ of avian- or human-adapted viruses. Restriction is hypothesized to result from differential interactions (either positive or inhibitory) with unidentified cellular factors. We applied fluorescence recovery after photobleaching (FRAP) to investigate the mobility of the viral polymerase in the cell nucleus using A/PR/8/34 and A/Turkey/England/50-92/91 as model strains. As expected, transcriptional activity of a polymerase with the avian PB2 protein was strongly dependent on the identity of residue 627 in human but not avian cells, and this correlated with significantly slower diffusion of the inactive polymerase in human but not avian nuclei. In contrast, the activity and mobility of the PR8 polymerase was affected much less by residue 627. Sequence comparison followed by mutagenic analyses identified residues at known host-range-specific positions 271, 588 and 701 as well as a novel determinant at position 636 as contributors to host-specific activity of both PR8 and Turkey PB2 proteins. Furthermore, the correlation between poor transcriptional activity and slow diffusional mobility was maintained. However, activity did not obligatorily correlate with predicted surface charge of the 627 domain. Overall, our data support the hypothesis of a host nuclear factor that interacts with the viral polymerase and modulates its activity. While we cannot distinguish between positive and inhibitory effects, the data have implications for how such factors might operate.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/vir.0.031492-0
2011-07-01
2024-03-28
Loading full text...

Full text loading...

/deliver/fulltext/jgv/92/7/1650.html?itemId=/content/journal/jgv/10.1099/vir.0.031492-0&mimeType=html&fmt=ahah

References

  1. Almond J. W. 1977; A single gene determines the host range of influenza virus. Nature 270:617–618 [View Article][PubMed]
    [Google Scholar]
  2. Benfield C. T., Lyall J. W., Kochs G., Tiley L. S. 2008; Asparagine 631 variants of the chicken Mx protein do not inhibit influenza virus replication in primary chicken embryo fibroblasts or in vitro surrogate assays. J Virol 82:7533–7539 [View Article][PubMed]
    [Google Scholar]
  3. Boivin S., Hart D. J. 2011; Interaction of the influenza A virus polymerase PB2 C-terminal region with importin α isoforms provides insights into host adaptation and polymerase assembly. J Biol Chem 286:10439–10448 [View Article][PubMed]
    [Google Scholar]
  4. Boivin S., Cusack S., Ruigrok R. W., Hart D. J. 2010; Influenza A virus polymerase: structural insights into replication and host adaptation mechanisms. J Biol Chem 285:28411–28417 [View Article][PubMed]
    [Google Scholar]
  5. Bussey K. A., Bousse T. L., Desmet E. A., Kim B., Takimoto T. 2010; PB2 residue 271 plays a key role in enhanced polymerase activity of influenza A viruses in mammalian host cells. J Virol 84:4395–4406 [View Article][PubMed]
    [Google Scholar]
  6. Carrasco M., Amorim M. J., Digard P. 2004; Lipid raft-dependent targeting of the influenza A virus nucleoprotein to the apical plasma membrane. Traffic 5:979–992 [View Article][PubMed]
    [Google Scholar]
  7. Chen G.-W., Chang S.-C., Mok C. K., Lo Y.-L., Kung Y.-N., Huang J.-H., Shih Y.-H., Wang J.-Y., Chiang C. et al. 2006; Genomic signatures of human versus avian influenza A viruses. Emerg Infect Dis 12:1353–1360[PubMed] [CrossRef]
    [Google Scholar]
  8. de Wit E., Spronken M. I. J., Bestebroer T. M., Rimmelzwaan G. F., Osterhaus A. D. M. E., Fouchier R. A. M. 2004; Efficient generation and growth of influenza virus A/PR/8/34 from eight cDNA fragments. Virus Res 103:155–161 [View Article][PubMed]
    [Google Scholar]
  9. Engelhardt O. G., Smith M., Fodor E. 2005; Association of the influenza A virus RNA-dependent RNA polymerase with cellular RNA polymerase II. J Virol 79:5812–5818 [View Article][PubMed]
    [Google Scholar]
  10. Fields S., Winter G. 1982; Nucleotide sequences of influenza virus segments 1 and 3 reveal mosaic structure of a small viral RNA segment. Cell 28:303–313 [View Article][PubMed]
    [Google Scholar]
  11. Finkelstein D. B., Mukatira S., Mehta P. K., Obenauer J. C., Su X., Webster R. G., Naeve C. W. 2007; Persistent host markers in pandemic and H5N1 influenza viruses. J Virol 81:10292–10299 [View Article][PubMed]
    [Google Scholar]
  12. Fodor E., Smith M. 2004; The PA subunit is required for efficient nuclear accumulation of the PB1 subunit of the influenza A virus RNA polymerase complex. J Virol 78:9144–9153 [View Article][PubMed]
    [Google Scholar]
  13. Gabriel G., Dauber B., Wolff T., Planz O., Klenk H.-D., Stech J. 2005; The viral polymerase mediates adaptation of an avian influenza virus to a mammalian host. Proc Natl Acad Sci U S A 102:18590–18595 [View Article][PubMed]
    [Google Scholar]
  14. Gabriel G., Herwig A., Klenk H. D. 2008; Interaction of polymerase subunit PB2 and NP with importin alpha1 is a determinant of host range of influenza A virus. PLoS Pathog 4:e11 [View Article][PubMed]
    [Google Scholar]
  15. Gao P., Watanabe S., Ito T., Goto H., Wells K., McGregor M., Cooley A. J., Kawaoka Y. 1999; Biological heterogeneity, including systemic replication in mice, of H5N1 influenza A virus isolates from humans in Hong Kong. J Virol 73:3184–3189[PubMed]
    [Google Scholar]
  16. Guilligay D., Tarendeau F., Resa-Infante P., Coloma R., Crepin T., Sehr P., Lewis J., Ruigrok R. W. H., Ortín J. et al. 2008; The structural basis for cap binding by influenza virus polymerase subunit PB2. Nat Struct Mol Biol 15:500–506 [View Article][PubMed]
    [Google Scholar]
  17. 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 [View Article][PubMed]
    [Google Scholar]
  18. Herfst S., Chutinimitkul S., Ye J., de Wit E., Munster V. J., Schrauwen E. J. A., Bestebroer T. M., Jonges M., Meijer A. et al. 2010; Introduction of virulence markers in PB2 of pandemic swine-origin influenza virus does not result in enhanced virulence or transmission. J Virol 84:3752–3758 [View Article][PubMed]
    [Google Scholar]
  19. Howard W., Hayman A., Lackenby A., Whiteley A., Londt B., Banks J., McCauley J. W., Barclay W. S. 2007; Development of a reverse genetics system enabling the rescue of recombinant avian influenza virus A/Turkey/England/50-92/91 (H5N1). Avian Dis 51:Suppl.393–395 [View Article][PubMed]
    [Google Scholar]
  20. Jagger B. W., Memoli M. J., Sheng Z.-M., Qi L., Hrabal R. J., Allen G. L., Dugan V. G., Wang R., Digard P. et al. 2010; The PB2-E627K mutation attenuates viruses containing the 2009 H1N1 influenza pandemic polymerase. mBio 1:e00067-10 [View Article][PubMed]
    [Google Scholar]
  21. Josset L., Frobert E., Rosa-Calatrava M. 2008; Influenza A replication and host nuclear compartments: many changes and many questions. J Clin Virol 43:381–390 [View Article][PubMed]
    [Google Scholar]
  22. Kuzuhara T., Kise D., Yoshida H., Horita T., Murazaki Y., Nishimura A., Echigo N., Utsunomiya H., Tsuge H. 2009; Structural basis of the influenza A virus RNA polymerase PB2 RNA-binding domain containing the pathogenicity-determinant lysine 627 residue. J Biol Chem 284:6855–6860 [View Article][PubMed]
    [Google Scholar]
  23. Labadie K., Dos Santos Afonso E., Rameix-Welti M.-A., van der Werf S., Naffakh N. 2007; Host-range determinants on the PB2 protein of influenza A viruses control the interaction between the viral polymerase and nucleoprotein in human cells. Virology 362:271–282 [View Article][PubMed]
    [Google Scholar]
  24. Le Q. M., Sakai-Tagawa Y., Ozawa M., Ito M., Kawaoka Y. 2009; Selection of H5N1 influenza virus PB2 during replication in humans. J Virol 83:5278–5281 [View Article][PubMed]
    [Google Scholar]
  25. Lee M. T. M., Bishop K., Medcalf L., Elton D., Digard P., Tiley L. S. 2002; Definition of the minimal viral components required for the initiation of unprimed RNA synthesis by influenza virus RNA polymerase. Nucleic Acids Res 30:429–438 [View Article][PubMed]
    [Google Scholar]
  26. Lee M.-T. M., Klumpp K., Digard P., Tiley L. S. 2003; Activation of influenza virus RNA polymerase by the 5′ and 3′ terminal duplex of genomic RNA. Nucleic Acids Res 31:1624–1632 [View Article][PubMed]
    [Google Scholar]
  27. Li Z., Chen H., Jiao P., Deng G., Tian G., Li Y., Hoffmann E., Webster R. G., Matsuoka Y., Yu K. 2005; Molecular basis of replication of duck H5N1 influenza viruses in a mammalian mouse model. J Virol 79:12058–12064 [View Article][PubMed]
    [Google Scholar]
  28. Li J., Ishaq M., Prudence M., Xi X., Hu T., Liu Q., Guo D. 2009; Single mutation at the amino acid position 627 of PB2 that leads to increased virulence of an H5N1 avian influenza virus during adaptation in mice can be compensated by multiple mutations at other sites of PB2. Virus Res 144:123–129 [View Article][PubMed]
    [Google Scholar]
  29. Loucaides E. M., von Kirchbach J. C., Foeglein A., Sharps J., Fodor E., Digard P. 2009; Nuclear dynamics of influenza A virus ribonucleoproteins revealed by live-cell imaging studies. Virology 394:154–163 [View Article][PubMed]
    [Google Scholar]
  30. Massin P., van der Werf S., Naffakh N. 2001; Residue 627 of PB2 is a determinant of cold sensitivity in RNA replication of avian influenza viruses. J Virol 75:5398–5404 [View Article][PubMed]
    [Google Scholar]
  31. Massin P., Kuntz-Simon G., Barbezange C., Deblanc C., Oger A., Marquet-Blouin E., Bougeard S., van der Werf S., Jestin V. 2010; Temperature sensitivity on growth and/or replication of H1N1, H1N2 and H3N2 influenza A viruses isolated from pigs and birds in mammalian cells. Vet Microbiol 142:232–241 [View Article][PubMed]
    [Google Scholar]
  32. Mehle A., Doudna J. A. 2008; An inhibitory activity in human cells restricts the function of an avian-like influenza virus polymerase. Cell Host Microbe 4:111–122 [View Article][PubMed]
    [Google Scholar]
  33. Mehle A., Doudna J. A. 2009; Adaptive strategies of the influenza virus polymerase for replication in humans. Proc Natl Acad Sci U S A 106:21312–21316 [View Article][PubMed]
    [Google Scholar]
  34. Miotto O., Heiny A., Tan T. W., August J. T., Brusic V. 2008; Identification of human-to-human transmissibility factors in PB2 proteins of influenza A by large-scale mutual information analysis. BMC Bioinformatics 9:Suppl. 1S18 [View Article][PubMed]
    [Google Scholar]
  35. Moncorgé O., Mura M., Barclay W. S. 2010; Evidence for avian and human host cell factors that affect the activity of influenza virus polymerase. J Virol 84:9978–9986 [View Article][PubMed]
    [Google Scholar]
  36. Mukaigawa J., Nayak D. P. 1991; Two signals mediate nuclear localization of influenza virus (A/WSN/33) polymerase basic protein 2. J Virol 65:245–253[PubMed]
    [Google Scholar]
  37. Mullin A. E., Dalton R. M., Amorim M. J., Elton D., Digard P. 2004; Increased amounts of the influenza virus nucleoprotein do not promote higher levels of viral genome replication. J Gen Virol 85:3689–3698 [View Article][PubMed]
    [Google Scholar]
  38. Munster V. J., de Wit E., van Riel D., Beyer W. E. P., Rimmelzwaan G. F., Osterhaus A. D. M. E., Kuiken T., Fouchier R. A. M. 2007; The molecular basis of the pathogenicity of the Dutch highly pathogenic human influenza A H7N7 viruses. J Infect Dis 196:258–265 [View Article][PubMed]
    [Google Scholar]
  39. Naffakh N., Massin P., Escriou N., Crescenzo-Chaigne B., van der Werf S. 2000; Genetic analysis of the compatibility between polymerase proteins from human and avian strains of influenza A viruses. J Gen Virol 81:1283–1291[PubMed]
    [Google Scholar]
  40. Naffakh N., Tomoiu A., Rameix-Welti M.-A., van der Werf S. 2008; Host restriction of avian influenza viruses at the level of the ribonucleoproteins. Annu Rev Microbiol 62:403–424 [View Article][PubMed]
    [Google Scholar]
  41. Neumann G., Brownlee G. G., Fodor E., Kawaoka Y. 2004; Orthomyxovirus replication, transcription, and polyadenylation. Curr Top Microbiol Immunol 283:121–143[PubMed]
    [Google Scholar]
  42. Phair R. D., Misteli T. 2000; High mobility of proteins in the mammalian cell nucleus. Nature 404:604–609 [View Article][PubMed]
    [Google Scholar]
  43. Poole E., Elton D., Medcalf L., Digard P. 2004; Functional domains of the influenza A virus PB2 protein: identification of NP- and PB1-binding sites. Virology 321:120–133 [View Article][PubMed]
    [Google Scholar]
  44. Portela A., Digard P. 2002; The influenza virus nucleoprotein: a multifunctional RNA-binding protein pivotal to virus replication. J Gen Virol 83:723–734[PubMed]
    [Google Scholar]
  45. Rameix-Welti M.-A., Tomoiu A., Dos Santos Afonso E., van der Werf S., Naffakh N. 2009; Avian influenza A virus polymerase association with nucleoprotein, but not polymerase assembly, is impaired in human cells during the course of infection. J Virol 83:1320–1331 [View Article][PubMed]
    [Google Scholar]
  46. Resa-Infante P., Jorba N., Zamarreño N., Fernández Y., Juárez S., Ortín J. 2008; The host-dependent interaction of alpha-importins with influenza PB2 polymerase subunit is required for virus RNA replication. PLoS ONE 3:e3904 [View Article][PubMed]
    [Google Scholar]
  47. Schnitzler S. U., Schnitzler P. 2009; An update on swine-origin influenza virus A/H1N1: a review. Virus Genes 39:279–292 [View Article][PubMed]
    [Google Scholar]
  48. Shinya K., Watanabe S., Ito T., Kasai N., Kawaoka Y. 2007; Adaptation of an H7N7 equine influenza A virus in mice. J Gen Virol 88:547–553 [View Article][PubMed]
    [Google Scholar]
  49. Steel J., Lowen A. C., Mubareka S., Palese P. 2009; Transmission of influenza virus in a mammalian host is increased by PB2 amino acids 627K or 627E/701N. PLoS Pathog 5:e1000252 [View Article][PubMed]
    [Google Scholar]
  50. Subbarao E. K., London W., Murphy B. R. 1993; A single amino acid in the PB2 gene of influenza A virus is a determinant of host range. J Virol 67:1761–1764[PubMed]
    [Google Scholar]
  51. Sugiyama K., Obayashi E., Kawaguchi A., Suzuki Y., Tame J. R., Nagata K., Park S. Y. 2009; Structural insight into the essential PB1-PB2 subunit contact of the influenza virus RNA polymerase. EMBO J 28:1803–1811 [View Article][PubMed]
    [Google Scholar]
  52. Tarendeau F., Boudet J., Guilligay D., Mas P. J., Bougault C. M., Boulo S., Baudin F., Ruigrok R. W. H., Daigle N. et al. 2007; Structure and nuclear import function of the C-terminal domain of influenza virus polymerase PB2 subunit. Nat Struct Mol Biol 14:229–233 [View Article][PubMed]
    [Google Scholar]
  53. Tarendeau F., Crepin T., Guilligay D., Ruigrok R. W. H., Cusack S., Hart D. J. 2008; Host determinant residue lysine 627 lies on the surface of a discrete, folded domain of influenza virus polymerase PB2 subunit. PLoS Pathog 4:e1000136 [View Article][PubMed]
    [Google Scholar]
  54. Taubenberger J. K., Kash J. C. 2010; Influenza virus evolution, host adaptation, and pandemic formation. Cell Host Microbe 7:440–451 [View Article][PubMed]
    [Google Scholar]
  55. Webster R. G., Bean W. J., Gorman O. T., Chambers T. M., Kawaoka Y. 1992; Evolution and ecology of influenza A viruses. Microbiol Rev 56:152–179[PubMed]
    [Google Scholar]
  56. Wise H. M., Foeglein A., Sun J., Dalton R. M., Patel S., Howard W., Anderson E. C., Barclay W. S., Digard P. 2009; A complicated message: identification of a novel PB1-related protein translated from influenza A virus segment 2 mRNA. J Virol 83:8021–8031 [View Article][PubMed]
    [Google Scholar]
  57. Wood G. W., McCauley J. W., Bashiruddin J. B., Alexander D. J. 1993; Deduced amino acid sequences at the haemagglutinin cleavage site of avian influenza A viruses of H5 and H7 subtypes. Arch Virol 130:209–217 [View Article][PubMed]
    [Google Scholar]
  58. Yamada S., Hatta M., Staker B. L., Watanabe S., Imai M., Shinya K., Sakai-Tagawa Y., Ito M., Ozawa M. et al. 2010; Biological and structural characterization of a host-adapting amino acid in influenza virus. PLoS Pathog 6:e1001034 [View Article][PubMed]
    [Google Scholar]
  59. Yao Y., Mingay L. J., McCauley J. W., Barclay W. S. 2001; Sequences in influenza A virus PB2 protein that determine productive infection for an avian influenza virus in mouse and human cell lines. J Virol 75:5410–5415 [View Article][PubMed]
    [Google Scholar]
  60. Zhu H., Wang J., Wang P., Song W., Zheng Z., Chen R., Guo K., Zhang T., Peiris J. S. M., Chen H. 2010; Substitution of lysine at 627 position in PB2 protein does not change virulence of the 2009 pandemic H1N1 virus in mice. Virology 401:1–5 [View Article][PubMed]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/vir.0.031492-0
Loading
/content/journal/jgv/10.1099/vir.0.031492-0
Loading

Data & Media loading...

Most cited Most Cited RSS feed