1887

Abstract

The influenza B virus genome RNA segment 7 encodes the M1 and BM2 proteins. The BM2 protein is synthesized by a coupled translational termination–reinitiation mechanism at the overlapping stop–start pentanucleotide in a bicistronic mRNA transcribed from RNA segment 7. However, features and functions of this protein remain unclear. In this study the BM2 protein was characterized by using an antiserum raised to the BM2 protein of influenza virus strain B/Yamagata/1/73. In cells infected with B/Yamagata virus the αBM2 antibody specifically detected the BM2 protein with a molecular mass of 12 kDa and also a polypeptide with a molecular mass of 17 kDa. When infected cells were labelled with P and immunoprecipitated with the αBM2 antibody, the P-labelled 17 kDa polypeptide was specifically precipitated. In the presence of casein kinase inhibitor CKI-7 the synthesis of the 17 kDa and BM2 proteins was completely suppressed, although other viral proteins, except for the polymerase protein, were synthesized normally. These results suggest that the 17 kDa species is a phosphorylated form of the BM2 protein. These species were substantially synthesized in the late phase of infection and localized in the cytoplasm throughout infection. Moreover, they were transported to the plasma membrane and thereafter were incorporated into virions. These results therefore suggest that the BM2 and the 17 kDa proteins are necessary for the life-cycle of influenza B virus.

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1999-10-01
2019-10-15
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References

  1. Almond, J. W. & Felsenreich, V. ( 1982; ). Phosphorylation of the nucleoprotein of an avian influenza virus. Journal of General Virology 60, 295-305.[CrossRef]
    [Google Scholar]
  2. Betakova, T. , Nermut, M. V. & Hay, A. J. ( 1996; ). The NB protein is an integral component of the membrane of influenza B virus. Journal of General Virology 77, 2689-2694 .[CrossRef]
    [Google Scholar]
  3. Brassard, D. L. , Leser, G. P. & Lamb, R. A. ( 1996; ). Influenza B virus NB glycoprotein is a component of the virion. Virology 220, 350-360.[CrossRef]
    [Google Scholar]
  4. Briedis, D. J. , Lamb, R. A. & Choppin, P. W. ( 1982; ). Sequence of RNA segment 7 of the influenza B virus genome: partial amino acid identity between the membrane proteins (M1) of influenza A and B viruses and conservation of a second open reading frame. Virology 116, 581-588.[CrossRef]
    [Google Scholar]
  5. Chijiwa, T. , Hagiwara, M. & Hidaka, H. ( 1989; ). A newly synthesized selective casein kinase I inhibitor, N-(2-aminoethyl)-5-chloroisoquinoline-8- sulfonamide, and affinity purification of casein kinase I from bovine testis. Journal of Biological Chemistry 264, 4924-4927 .
    [Google Scholar]
  6. Enami, K. , Qiao, Y. , Fukuda, R. & Enami, M. ( 1993; ). An influenza virus temperature- sensitive mutant defective in the nuclear–cytoplasmic transport of the negative-sense viral RNAs. Virology 194, 822-827.[CrossRef]
    [Google Scholar]
  7. Gregoriades, A. , Christie, T. & Markarian, K. ( 1984; ). The membrane (M1) protein of influenza virus occurs in two forms and is a phosphoprotein. Journal of Virology 49, 229-235.
    [Google Scholar]
  8. Hardie, D. G. (1993). Protein Phosphorylation: A Practical Approach, 1st edn. Edited by D. G. Hardie. Oxford: Oxford University Press.
  9. Hiebert, S. W. , Williams, M. A. & Lamb, R. A. ( 1986; ). Nucleotide sequence of RNA segment 7 of influenza B/Singapore/222/79: maintenance of a second large open reading frame. Virology 155, 747-751.[CrossRef]
    [Google Scholar]
  10. Holsinger, L. J. , Nichani, D. , Pinto, L. H. & Lamb, R. A. ( 1994; ). Influenza A virus M2 ion channel protein: a structure–function analysis. Journal of Virology 68, 1551-1563 .
    [Google Scholar]
  11. Horvath, C. M. , Williams, M. A. & Lamb, R. A. ( 1990; ). Eukaryotic coupled translation of tandem cistrons: identification of the influenza B virus BM2 polypeptide. EMBO Journal 9, 2639-2647.
    [Google Scholar]
  12. Kozak, M. ( 1983; ). Comparison of initiation of protein synthesis in procaryotes, eucaryotes, and organelles. Microbiological Review 47, 1-45.
    [Google Scholar]
  13. Krantz, D. E. , Peter, D. , Liu, Y. & Edwards, R. H. ( 1997; ). Phosphorylation of a vesicular monoamine transporter by casein kinase II. Journal of Biological Chemistry 272, 6752-6759 .[CrossRef]
    [Google Scholar]
  14. Lamb, R. A. ( 1989; ). The influenza viruses. In Genes and Proteins of the Influenza Viruses, pp. 1-87. Edited by R. M. Krug. NY: Plenum Press.
  15. Lamb, R. A. & Krug, R. M. (1996). Orthomyxoviridae: the viruses and their replication. Fields Virology, 3rd edn, pp. 1353–1395. Edited by B. N. Fields, D. M. Knipe & P. M. Howley. Philadelphia & New York: Lippincott–Raven.
  16. 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. Proceedings of the National Academy of Sciences, USA 78, 4170-4174 .[CrossRef]
    [Google Scholar]
  17. Martin, K. & Helenius, A. ( 1991; ). Nuclear transport of influenza virus ribonucleoproteins: the viral matrix protein (M1) promotes export and inhibits import. Cell 67, 117-130.[CrossRef]
    [Google Scholar]
  18. Nakagawa, N. , Maeda, A. , Kase, T. , Kubota, R. & Okuno, Y. ( 1999; ). Rapid detection and identification of two lineages of influenza B strains with monoclonal antibodies. Journal of Virological Methods 79, 113-120.[CrossRef]
    [Google Scholar]
  19. Neumann, G. , Castrucci, M. R. & Kawaoka, Y. ( 1997; ). Nuclear import and export of influenza virus nucleoprotein. Journal of Virology 71, 9690-9700 .
    [Google Scholar]
  20. Nigg, E. A. ( 1997; ). Nucleocytoplasmic transport: signals, mechanisms and regulation. Nature 386, 779-787.[CrossRef]
    [Google Scholar]
  21. Niwa, H. , Yamamura, K. & Miyazaki, J. ( 1991; ). Efficient selection for high- expression transfectants with a novel eukaryotic vector. Gene 108, 193-199.[CrossRef]
    [Google Scholar]
  22. Odagiri, T. & Tashiro, M. ( 1997; ). Segment-specific noncoding sequences of the influenza virus genome RNA are involved in the specific competition between defective interfering RNA and its progenitor RNA segment at the virion assembly step. Journal of Virology 71, 2138-2145 .
    [Google Scholar]
  23. Odagiri, T. & Tobita, K. ( 1990; ). Mutation in NS2, a nonstructural protein of influenza A virus, extragenically causes aberrant replication and expression of the PA gene and leads to generation of defective interfering particles. Proceedings of the National Academy of Sciences, USA 87, 5988-5992 .[CrossRef]
    [Google Scholar]
  24. Odagiri, T. , DeBorde, D. C. & Maassab, H. F. ( 1982; ). Cold-adapted recombinants of influenza A virus in MDCK cells. I. Development and characterization of A/Ann Arbor/6/60×A/Alaska/6/77 recombinant viruses. Virology 119, 82-95.[CrossRef]
    [Google Scholar]
  25. O’Neill, R. E. , Talon, J. & Palese, P. ( 1998; ). The influenza virus NEP (NS2 protein) mediates the nuclear export of viral ribonucleoproteins. EMBO Journal 17, 288-296.[CrossRef]
    [Google Scholar]
  26. Parvin, J. D. , Palese, P. , Honda, A. , Ishihama, A. & Krystal, M. ( 1989; ). Promoter analysis of influenza virus RNA polymerase. Journal of Virology 63, 5142-5152 .
    [Google Scholar]
  27. Peluso, R. W. , Lamb, R. A. & Choppin, P. W. ( 1977; ). Polypeptide synthesis in simian virus 5-infected cells. Journal of Virology 23, 177-187.
    [Google Scholar]
  28. Pinto, L. H. , Holsinger, L. J. & Lamb, R. A. ( 1992; ). Influenza virus M2 protein has ion channel activity. Cell 69, 517-528.[CrossRef]
    [Google Scholar]
  29. Shaw, M. W. , Choppin, P. W. & Lamb, R. A. ( 1983; ). A previously unrecognized influenza B virus glycoprotein from a bicistronic mRNA that also encodes the viral neuraminidase. Proceedings of the National Academy of Sciences, USA 80, 4879-4883 .[CrossRef]
    [Google Scholar]
  30. Steinhauer, D. A. , Wharton, S. A. , Skehel, J. J. , Wiley, D. C. & Hay, A. J. ( 1991; ). Amantadine selection of a mutant influenza virus containing an acid-stable hemagglutinin glycoprotein: evidence for virus-specific regulation of the pH of glycoprotein transport vesicles. Proceedings of the National Academy of Sciences, USA 88, 11525-11529 .[CrossRef]
    [Google Scholar]
  31. Sunstrom, N. A. , Premkumar, L. S. , Premkumar, A. , Ewart, G. , Cox, G. B. & Gage, P. W. ( 1996; ). Ion channels formed by NB, an influenza B virus protein. Journal of Membrane Biology 150, 127-132.[CrossRef]
    [Google Scholar]
  32. Wang, P. , Palese, P. & O’Neill, R. E. ( 1997; ). The NPI-1/NPI- 3 (karyopherin alpha) binding site on the influenza A virus nucleoprotein NP is a nonconventional nuclear localization signal. Journal of Virology 71, 1850-1856 .
    [Google Scholar]
  33. Ward, A. C. , Castelli, L. A. , Lucantoni, A. C. , White, J. F. , Azad, A. A. & Macreadie, I. G. ( 1995; ). Expression and analysis of the NS2 protein of influenza A virus. Archives of Virology 140, 2067-2073 .[CrossRef]
    [Google Scholar]
  34. Whittaker, G. , Kemler, I. & Helenius, A. ( 1995; ). Hyperphosphorylation of mutant influenza virus matrix protein, M1, causes its retention in the nucleus. Journal of Virology 69, 439-445.
    [Google Scholar]
  35. Williams, M. A. & Lamb, R. A. ( 1989; ). Effect of mutations and deletions in a bicistronic mRNA on the synthesis of influenza B virus NB and NA glycoproteins. Journal of Virology 63, 28-35.
    [Google Scholar]
  36. Yasuda, J. , Nakada, S. , Kato, A. , Toyoda, T. & Ishihama, A. ( 1993; ). Molecular assembly of influenza virus: association of the NS2 protein with virion matrix. Virology 196, 249-255.[CrossRef]
    [Google Scholar]
  37. Ye, Z. , Baylor, N. W. & Wagner, R. R. ( 1989; ). Transcription- inhibition and RNA-binding domains of influenza A virus matrix protein mapped with anti-idiotypic antibodies and synthetic peptides. Journal of Virology 63, 3586-3594 .
    [Google Scholar]
  38. Zhirnov, O. P. & Bukrinskaya, A. G. ( 1981; ). Two forms of influenza virus nucleoprotein in infected cells and virions. Virology 109, 174-179.[CrossRef]
    [Google Scholar]
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