1887

Abstract

The influenza A virus M1 mRNA is alternatively spliced to produce M2 mRNA, mRNA, and in some cases, M4 mRNA. Splicing of influenza mRNAs is carried out by the cellular splicing machinery and is thought to be regulated, as both spliced and unspliced mRNAs encode proteins. In this study, we used radioactively labelled primers to investigate the accumulation of spliced and unspliced M segment mRNAs in viral infection and ribonucleoprotein (RNP) reconstitution assays in which only the minimal components required for transcription and replication to occur were expressed. We found that co-expression of the viral NS1 protein in an RNP reconstitution assay altered the accumulation of spliced mRNAs compared with when it was absent, and that this activity was dependent on the RNA-binding ability of NS1. These findings suggest that the NS1 protein plays a role in the regulation of splicing of influenza virus M1 mRNA.

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2012-01-01
2019-10-18
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References

  1. Alonso-Caplen F. V., Krug R. M.. ( 1991;). Regulation of the extent of splicing of influenza virus NS1 mRNA: role of the rates of splicing and of the nucleocytoplasmic transport of NS1 mRNA. . Mol Cell Biol 11:, 1092–1098.[PubMed]
    [Google Scholar]
  2. Bier K., York A., Fodor E.. ( 2011;). Cellular cap-binding proteins associate with influenza virus mRNAs. . J Gen Virol 92:, 1627–1634. [CrossRef][PubMed]
    [Google Scholar]
  3. Chen Z., Krug R. M.. ( 2000;). Selective nuclear export of viral mRNAs in influenza-virus-infected cells. . Trends Microbiol 8:, 376–383. [CrossRef][PubMed]
    [Google Scholar]
  4. Chen Z., Li Y., Krug R. M.. ( 1999;). Influenza A virus NS1 protein targets poly(A)-binding protein II of the cellular 3′-end processing machinery. . EMBO J 18:, 2273–2283. [CrossRef][PubMed]
    [Google Scholar]
  5. Engelhardt O. G., Fodor E.. ( 2006;). Functional association between viral and cellular transcription during influenza virus infection. . Rev Med Virol 16:, 329–345. [CrossRef][PubMed]
    [Google Scholar]
  6. 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]
  7. 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. [CrossRef][PubMed]
    [Google Scholar]
  8. 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]
  9. Garaigorta U., Ortín J.. ( 2007;). Mutation analysis of a recombinant NS replicon shows that influenza virus NS1 protein blocks the splicing and nucleo-cytoplasmic transport of its own viral mRNA. . Nucleic Acids Res 35:, 4573–4582. [CrossRef][PubMed]
    [Google Scholar]
  10. Ge H., Manley J. L.. ( 1990;). A protein factor, ASF, controls cell-specific alternative splicing of SV40 early pre-mRNA in vitro. . Cell 62:, 25–34. [CrossRef][PubMed]
    [Google Scholar]
  11. 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. [CrossRef][PubMed]
    [Google Scholar]
  12. Hatada E., Fukuda R.. ( 1992;). Binding of influenza A virus NS1 protein to dsRNA in vitro. . J Gen Virol 73:, 3325–3329. [CrossRef][PubMed]
    [Google Scholar]
  13. Hatada E., Takizawa T., Fukuda R.. ( 1992;). Specific binding of influenza A virus NS1 protein to the virus minus-sense RNA in vitro. . J Gen Virol 73:, 17–25. [CrossRef][PubMed]
    [Google Scholar]
  14. Hatada E., Saito S., Okishio N., Fukuda R.. ( 1997;). Binding of the influenza virus NS1 protein to model genome RNAs. . J Gen Virol 78:, 1059–1063.[PubMed]
    [Google Scholar]
  15. Lamb R. A., Choppin P. W.. ( 1981;). Identification of a second protein (M2) encoded by RNA segment 7 of influenza virus. . Virology 112:, 729–737. [CrossRef][PubMed]
    [Google Scholar]
  16. Lamb R. A., Lai C. J.. ( 1982;). Spliced and unspliced messenger RNAs synthesized from cloned influenza virus M DNA in an SV40 vector: expression of the influenza virus membrane protein (M1). . Virology 123:, 237–256. [CrossRef][PubMed]
    [Google Scholar]
  17. Lamb R. A., Lai C. J., Choppin P. W.. ( 1981;). Sequences of mRNAs derived from genome RNA segment 7 of influenza virus: colinear and interrupted mRNAs code for overlapping proteins. . Proc Natl Acad Sci U S A 78:, 4170–4174. [CrossRef][PubMed]
    [Google Scholar]
  18. 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. [CrossRef][PubMed]
    [Google Scholar]
  19. Lu Y., Wambach M., Katze M. G., Krug R. M.. ( 1995;). Binding of the influenza virus NS1 protein to double-stranded RNA inhibits the activation of the protein kinase that phosphorylates the elF-2 translation initiation factor. . Virology 214:, 222–228. [CrossRef][PubMed]
    [Google Scholar]
  20. Marión R. M., Aragón T., Beloso A., Nieto A., Ortín J.. ( 1997;). The N-terminal half of the influenza virus NS1 protein is sufficient for nuclear retention of mRNA and enhancement of viral mRNA translation. . Nucleic Acids Res 25:, 4271–4277. [CrossRef][PubMed]
    [Google Scholar]
  21. Mount S. M.. ( 1982;). A catalogue of splice junction sequences. . Nucleic Acids Res 10:, 459–472. [CrossRef][PubMed]
    [Google Scholar]
  22. Nemeroff M. E., Barabino S. M., Li Y., Keller W., Krug R. M.. ( 1998;). Influenza virus NS1 protein interacts with the cellular 30 kDa subunit of CPSF and inhibits 3’end formation of cellular pre-mRNAs. . Mol Cell 1:, 991–1000. [CrossRef][PubMed]
    [Google Scholar]
  23. Qiu Y., Krug R. M.. ( 1994;). The influenza virus NS1 protein is a poly(A)-binding protein that inhibits nuclear export of mRNAs containing poly(A). . J Virol 68:, 2425–2432.[PubMed]
    [Google Scholar]
  24. Qiu Y., Nemeroff M., Krug R. M.. ( 1995;). The influenza virus NS1 protein binds to a specific region in human U6 snRNA and inhibits U6-U2 and U6-U4 snRNA interactions during splicing. . RNA 1:, 304–316.[PubMed]
    [Google Scholar]
  25. Robb N. C., Jackson D., Vreede F. T., Fodor E.. ( 2010;). Splicing of influenza A virus NS1 mRNA is independent of the viral NS1 protein. . J Gen Virol 91:, 2331–2340. [CrossRef][PubMed]
    [Google Scholar]
  26. Robb N. C., Chase G., Bier K., Vreede F. T., Shaw P. C., Naffakh N., Schwemmle M., Fodor E.. ( 2011;). The influenza A virus NS1 protein interacts with the nucleoprotein of viral ribonucleoprotein complexes. . J Virol 85:, 5228–5231. [CrossRef][PubMed]
    [Google Scholar]
  27. 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. [CrossRef][PubMed]
    [Google Scholar]
  28. Satterly N., Tsai P. L., van Deursen J., Nussenzveig D. R., Wang Y., Faria P. A., Levay A., Levy D. E., Fontoura B. M.. ( 2007;). Influenza virus targets the mRNA export machinery and the nuclear pore complex. . Proc Natl Acad Sci U S A 104:, 1853–1858. [CrossRef][PubMed]
    [Google Scholar]
  29. 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. [CrossRef][PubMed]
    [Google Scholar]
  30. Shih S. R., Krug R. M.. ( 1996;). Novel exploitation of a nuclear function by influenza virus: the cellular SF2/ASF splicing factor controls the amount of the essential viral M2 ion channel protein in infected cells. . EMBO J 15:, 5415–5427.[PubMed]
    [Google Scholar]
  31. 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. [CrossRef][PubMed]
    [Google Scholar]
  32. Shih S. R., Suen P. C., Chen Y. S., Chang S. C.. ( 1998;). A novel spliced transcript of influenza A/WSN/33 virus. . Virus Genes 17:, 179–183. [CrossRef][PubMed]
    [Google Scholar]
  33. Valcárcel J., Portela A., Ortín J.. ( 1991;). Regulated M1 mRNA splicing in influenza virus-infected cells. . J Gen Virol 72:, 1301–1308. [CrossRef][PubMed]
    [Google Scholar]
  34. Wang W., Riedel K., Lynch P., Chien C. Y., Montelione G. T., Krug R. M.. ( 1999;). RNA binding by the novel helical domain of the influenza virus NS1 protein requires its dimer structure and a small number of specific basic amino acids. . RNA 5:, 195–205. [CrossRef][PubMed]
    [Google Scholar]
  35. 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. [CrossRef][PubMed]
    [Google Scholar]
  36. Winter G., Fields S.. ( 1980;). Cloning of influenza cDNA into M13: the sequence of the RNA segment encoding the A/PR/8/34 matrix protein. . Nucleic Acids Res 8:, 1965–1974. [CrossRef][PubMed]
    [Google Scholar]
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