1887

Abstract

The influenza virus RNA polymerase synthesizes three types of RNA: genomic vRNA, anti-genomic cRNA and mRNA. Both vRNA and cRNA are bound by the viral RNA polymerase and nucleoprotein to form ribonucleoprotein complexes. Viral mRNAs are also proposed to be bound by the RNA polymerase to prevent their endonucleolytic cleavage, regulate the splicing of M1 mRNA, and facilitate translation. Here, we used standard immunoprecipitation, biochemical purification and RNA immunoprecipitation assays to investigate the association of viral and host factors with viral mRNA. We found that viral mRNA associates with the viral non-structural protein 1 (NS1), cellular poly(A)-binding protein 1 (PABP1), the 20 kDa subunit NCBP1 of the nuclear cap-binding complex (CBC), the RNA and export factor-binding protein REF/Aly and the translation initiation factor eIF4E. However, our data suggest that the RNA polymerase might not form part of the viral messenger ribonucleoprotein (mRNP) complex. We propose a model in which viral mRNAs, by associating with cellular cap-binding proteins, follow the pathways normally used by cellular mRNAs for splicing, nuclear export and translation.

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2011-07-01
2024-03-29
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References

  1. Aguilera A. 2005; Cotranscriptional mRNP assembly: from the DNA to the nuclear pore. Curr Opin Cell Biol 17:242–250 [View Article][PubMed]
    [Google Scholar]
  2. Amorim M. J., Digard P. 2006; Influenza A virus and the cell nucleus. Vaccine 24:6651–6655 [View Article][PubMed]
    [Google Scholar]
  3. Aragón T., de la Luna S., Novoa I., Carrasco L., Ortín J., Nieto A. 2000; Eukaryotic translation initiation factor 4GI is a cellular target for NS1 protein, a translational activator of influenza virus. Mol Cell Biol 20:6259–6268 [View Article][PubMed]
    [Google Scholar]
  4. Braam J., Ulmanen I., Krug R. M. 1983; Molecular model of a eucaryotic transcription complex: functions and movements of influenza P proteins during capped RNA-primed transcription. Cell 34:611–618 [View Article][PubMed]
    [Google Scholar]
  5. Burgui I., Aragón T., Ortín J., Nieto A. 2003; PABP1 and eIF4GI associate with influenza virus NS1 protein in viral mRNA translation initiation complexes. J Gen Virol 84:3263–3274 [View Article][PubMed]
    [Google Scholar]
  6. Burgui I., Yángüez E., Sonenberg N., Nieto A. 2007; Influenza virus mRNA translation revisited: is the eIF4E cap-binding factor required for viral mRNA translation?. J Virol 81:12427–12438 [View Article][PubMed]
    [Google Scholar]
  7. Chan A. Y., Vreede F. T., Smith M., Engelhardt O. G., Fodor E. 2006; Influenza virus inhibits RNA polymerase II elongation. Virology 351:210–217 [View Article][PubMed]
    [Google Scholar]
  8. de la Luna S., Fortes P., Beloso A., Ortín J. 1995; Influenza virus NS1 protein enhances the rate of translation initiation of viral mRNAs. J Virol 69:2427–2433[PubMed]
    [Google Scholar]
  9. Deng T., Sharps J., Fodor E., Brownlee G. G. 2005; In vitro assembly of PB2 with a PB1-PA dimer supports a new model of assembly of influenza A virus polymerase subunits into a functional trimeric complex. J Virol 79:8669–8674 [View Article][PubMed]
    [Google Scholar]
  10. Dias A., Bouvier D., Crépin T., McCarthy A. A., Hart D. J., Baudin F., Cusack S., Ruigrok R. W. 2009; The cap-snatching endonuclease of influenza virus polymerase resides in the PA subunit. Nature 458:914–918 [View Article][PubMed]
    [Google Scholar]
  11. Enami K., Sato T. A., Nakada S., Enami M. 1994; Influenza virus NS1 protein stimulates translation of the M1 protein. J Virol 68:1432–1437[PubMed]
    [Google Scholar]
  12. Engelhardt O. G., Fodor E. 2006; Functional association between viral and cellular transcription during influenza virus infection. Rev Med Virol 16:329–345 [View Article][PubMed]
    [Google Scholar]
  13. Fechter P., Mingay L., Sharps J., Chambers A., Fodor E., Brownlee G. G. 2003; Two aromatic residues in the PB2 subunit of influenza A RNA polymerase are crucial for cap binding. J Biol Chem 278:20381–20388 [View Article][PubMed]
    [Google Scholar]
  14. 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]
  15. Fodor E., Devenish L., Engelhardt O. G., Palese P., Brownlee G. G., García-Sastre A. 1999; Rescue of influenza A virus from recombinant DNA. J Virol 73:9679–9682[PubMed]
    [Google Scholar]
  16. Fodor E., Crow M., Mingay L. J., Deng T., Sharps J., Fechter P., Brownlee G. G. 2002; A single amino acid mutation in the PA subunit of the influenza virus RNA polymerase inhibits endonucleolytic cleavage of capped RNAs. J Virol 76:8989–9001 [View Article][PubMed]
    [Google Scholar]
  17. Fortes P., Beloso A., Ortín J. 1994; Influenza virus NS1 protein inhibits pre-mRNA splicing and blocks mRNA nucleocytoplasmic transport. EMBO J 13:704–712[PubMed]
    [Google Scholar]
  18. Guilligay D., Tarendeau F., Resa-Infante P., Coloma R., Crepin T., Sehr P., Lewis J., Ruigrok R. W., Ortin 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]
  19. Hale B. G., Randall R. E., Ortín J., Jackson D. 2008; The multifunctional NS1 protein of influenza A viruses. J Gen Virol 89:2359–2376 [View Article][PubMed]
    [Google Scholar]
  20. Hara K., Schmidt F. I., Crow M., Brownlee G. G. 2006; Amino acid residues in the N-terminal region of the PA subunit of influenza A virus RNA polymerase play a critical role in protein stability, endonuclease activity, cap binding, and virion RNA promoter binding. J Virol 80:7789–7798 [View Article][PubMed]
    [Google Scholar]
  21. Lu Y., Qian X. Y., Krug R. M. 1994; The influenza virus NS1 protein: a novel inhibitor of pre-mRNA splicing. Genes Dev 8:1817–1828 [View Article][PubMed]
    [Google Scholar]
  22. Marión R. M., Zürcher T., de la Luna S., Ortín J. 1997; Influenza virus NS1 protein interacts with viral transcription-replication complexes in vivo . J Gen Virol 78:2447–2451[PubMed]
    [Google Scholar]
  23. Muraki Y., Furukawa T., Kohno Y., Matsuzaki Y., Takashita E., Sugawara K., Hongo S. 2010; Influenza C virus NS1 protein upregulates the splicing of viral mRNAs. J Virol 84:1957–1966 [View Article][PubMed]
    [Google Scholar]
  24. 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]
  25. Palese P., Shaw M. L. 2007; Orthomyxoviridae: the viruses and their replication. In Fields Virology pp. 1647–1689 Edited by Knipe D. M., Howley P. M. Philadelphia: Wolters Kluwer and Lippincott Williams & Wilkins;
    [Google Scholar]
  26. Park Y. W., Katze M. G. 1995; Translational control by influenza virus. Identification of cis-acting sequences and trans-acting factors which may regulate selective viral mRNA translation. J Biol Chem 270:28433–28439[PubMed] [CrossRef]
    [Google Scholar]
  27. Pleschka S., Jaskunas R., Engelhardt O. G., Zürcher T., Palese P., García-Sastre A. 1996; A plasmid-based reverse genetics system for influenza A virus. J Virol 70:4188–4192[PubMed]
    [Google Scholar]
  28. Poon L. L., Pritlove D. C., Fodor E., Brownlee G. G. 1999; Direct evidence that the poly(A) tail of influenza A virus mRNA is synthesized by reiterative copying of a U track in the virion RNA template. J Virol 73:3473–3476[PubMed]
    [Google Scholar]
  29. Read E. K., Digard P. 2010; Individual influenza A virus mRNAs show differential dependence on cellular NXF1/TAP for their nuclear export. J Gen Virol 91:1290–1301 [View Article][PubMed]
    [Google Scholar]
  30. Robb N. C., Smith M., Vreede F. T., Fodor E. 2009; NS2/NEP protein regulates transcription and replication of the influenza virus RNA genome. J Gen Virol 90:1398–1407 [View Article][PubMed]
    [Google Scholar]
  31. Ruigrok R. W., Crépin T., Hart D. J., Cusack S. 2010; Towards an atomic resolution understanding of the influenza virus replication machinery. Curr Opin Struct Biol 20:104–113 [View Article][PubMed]
    [Google Scholar]
  32. Salvatore M., Basler C. F., Parisien J. P., Horvath C. M., Bourmakina S., Zheng H., Muster T., Palese P., García-Sastre A. 2002; Effects of influenza A virus NS1 protein on protein expression: the NS1 protein enhances translation and is not required for shutoff of host protein synthesis. J Virol 76:1206–1212 [View Article][PubMed]
    [Google Scholar]
  33. Schneider J., Wolff T. 2009; Nuclear functions of the influenza A and B viruses NS1 proteins: do they play a role in viral mRNA export?. Vaccine 27:6312–6316 [View Article][PubMed]
    [Google Scholar]
  34. Shih S. R., Krug R. M. 1996; Surprising function of the three influenza viral polymerase proteins: selective protection of viral mRNAs against the cap-snatching reaction catalyzed by the same polymerase proteins. Virology 226:430–435 [View Article][PubMed]
    [Google Scholar]
  35. Shih S. R., Nemeroff M. E., Krug R. M. 1995; The choice of alternative 5′ splice sites in influenza virus M1 mRNA is regulated by the viral polymerase complex. Proc Natl Acad Sci U S A 92:6324–6328 [View Article][PubMed]
    [Google Scholar]
  36. Topisirovic I., Svitkin Y. V., Sonenberg N., Shatkin A. J. 2011; Cap and cap-binding proteins in the control of gene expression. Wiley Interdisciplinary Reviews: RNA 2:277–298 [View Article]
    [Google Scholar]
  37. Valcárcel J., Fortes P., Ortín J. 1993; Splicing of influenza virus matrix protein mRNA expressed from a simian virus 40 recombinant. J Gen Virol 74:1317–1326 [View Article][PubMed]
    [Google Scholar]
  38. Vreede F. T., Chan A. Y., Sharps J., Fodor E. 2010; Mechanisms and functional implications of the degradation of host RNA polymerase II in influenza virus infected cells. Virology 396:125–134 [View Article][PubMed]
    [Google Scholar]
  39. Wang W., Cui Z. Q., Han H., Zhang Z. P., Wei H. P., Zhou Y. F., Chen Z., Zhang X. E. 2008; Imaging and characterizing influenza A virus mRNA transport in living cells. Nucleic Acids Res 36:4913–4928 [View Article][PubMed]
    [Google Scholar]
  40. Watanabe T., Watanabe S., Kawaoka Y. 2010; Cellular networks involved in the influenza virus life cycle. Cell Host Microbe 7:427–439 [View Article][PubMed]
    [Google Scholar]
  41. West S., Gromak N., Proudfoot N. J. 2004; Human 5′→3′ exonuclease Xrn2 promotes transcription termination at co-transcriptional cleavage sites. Nature 432:522–525 [View Article][PubMed]
    [Google Scholar]
  42. Yuan P., Bartlam M., Lou Z., Chen S., Zhou J., He X., Lv Z., Ge R., Li X. et al. 2009; Crystal structure of an avian influenza polymerase PA(N) reveals an endonuclease active site. Nature 458:909–913 [View Article][PubMed]
    [Google Scholar]
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