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1887

Journal of General Virology header logo Journal of General Virology

Volume 71, Issue 6

Analysis of influenza A virus temperature-sensitive mutants with mutations in RNA segment 8 No Access

Abstract

Temperature-sensitive (ts) mutants of influenza virus strain A/Udorn/72 (H3N2 subtype) with lesions in RNA segment 8 exhibited intrasegmental complementation, and were divided in two complementation groups (HI and H2) on MDCK cells. The nucleotide sequence of segment 8 was determined for three of these mutants. The HI strains, ICR1629 and SPC45, have a single amino acid substitution in the coding region of the non-structural protein NS1, whereas the H2 strain, ICR516, has a substitution in the NS2- coding region. With both NS1 ts mutants, the synthesis of two late proteins, the matrix protein (Ml) and haemagglutinin (HA), was greatly reduced and NS1 synthesis also decreased at 40 °C (non-permissive temperature) compared to that at 34 °C (permissive temperature). The synthesis of each virus-specific RNA was analysed using a quantitative hybridization method. However, at 40 °C, the levels of individual mRNAs including those for the late proteins, were almost the same as those at 34 °C, and attained the wild-type levels later in the infection (5 h postinfection) when the synthesis of the late proteins and the NS1 protein was severely reduced. The observations suggest that the NS 1 protein, which is a nuclear protein, is involved in some post-transcriptional processes in the synthesis of the late proteins and the NS1 protein.

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© Society for General Microbiology 1990
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1990-06-01
2025-03-15
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References

  1. Almond J. W., Mcgeoch D., Barry R. D. 1979; Temperature-sensitive mutants of fowl plague virus: isolation and genetic characterization. Virology 92:416–427
     [Google Scholar]
  2. Berk A. J., Sharp P. A. 1978; Spliced early mRNAs of simian virus 40. Proceedings of the National Academy of Sciences, U.S.A 75:1274–1278
     [Google Scholar]
  3. Briedis D. J., Lamb R. A. 1981; Migration of influenza virus-specific polypeptides from cytoplasm to nucleus of infected cells. Virology 111:154–164
     [Google Scholar]
  4. Buonagurio D. A., Krystal M., Palese P., Deborde D. C., Maassab H. F. 1984; Analysis of an influenza A virus mutant with a deletion in the NS segment. Journal of Virology 49:418–425
     [Google Scholar]
  5. Bürger H., Steuler H., Scholtissek C. 1985; A mutant of fowl plague virus (influenza A) with an enhanced electrophoretic mobility of RNA segment 8. Journal of General Virology 66:1679–1686
     [Google Scholar]
  6. Chou P. Y., Fasman G. D. 1978; Empirical predictions of protein conformation. Annual Review of Biochemistry 47:251–276
     [Google Scholar]
  7. Davey J., Colman A., Dimmock N. J. 1985; Location of influenza virus M, NP and NS1 proteins in microinjected cells. Journal of General Virology 66:2319–2334
     [Google Scholar]
  8. Enami M., Fukuda R., Ishihama A. 1985; Transcription and replication of eight RNA segments of influenza virus. Virology 142:68–77
     [Google Scholar]
  9. Feramisco J. R., Smart J. E., Burridge K., Heleman D. M., Thomas G. P. 1982; Coexistence of vinculin-like protein of higher molecular weight in smooth muscle. Journal of Biological Chemistry 257:11024–11031
     [Google Scholar]
  10. Greenspan D., Palese P., Krystal M. 1988; Two nuclear location signals in the influenza virus NSI non structural protein. Journal of Virology 62:3020–3026
     [Google Scholar]
  11. Hatada E., Hasegawa M., Mukaigawa J., Shimizu K., Fukuda R. 1989; Control of influenza virus gene expression: quantitative analysis of each viral RNA species in infected cells. Journal of Biochemistry 105:537–546
     [Google Scholar]
  12. Inglis S. C., Mahy B. W. J. 1979; Polypeptides specified by the influenza genome. 3. Control of synthesis in infected cells. Virology 95:154–164
     [Google Scholar]
  13. Inglis S. C., Barrett T., Brown C. M., Almond J. W. 1979; The smallest genome RNA segment of influenza virus contains two genes that may overlap. Proceedings of the National Academy of Sciences, U.S.A 76:3790–3794
     [Google Scholar]
  14. Inglis S. C., Gething M.-J., Brown C. M. 1980; Relationship between the messenger RNAs transcribed from two overlapping genes of influenza virus. Nucleic Acids Research 8:3575–3589
     [Google Scholar]
  15. Katze M. G., Detjen B. M., Safer B., Krug R. M. 1986; Translational control by influenza virus: suppression of the kinase that phosphorylates the alpha subunit of initiation factor eIF-2 and selective translation of influenza viral messenger RNAs. Molecular and Cellular Biology 6:1741–1750
     [Google Scholar]
  16. Katze M. G., Tomita J., Black T., Krug R. M., Safer B., Hovanessian A. 1988; Influenza virus regulates protein synthesis during infection by repressing autophosphorylation and activity of the cellular 68,000-Mr protein kinase. Journal of Virology 62:3710–3717
     [Google Scholar]
  17. Koennecke I., Boschek C. B., Scholtissek C. 1981; Isolation and properties of a temperature-sensitive mutant (ts412) of an influenza A virus recombinant with a ts lesion in the gene coding for the nonstructural protein. Virology 110:16–25
     [Google Scholar]
  18. Krug R. M., Etkind P. R. 1973; Cytoplasmic and nuclear virus-specific proteins in influenza virus-infected MDCK cells. Virology 56:334–348
     [Google Scholar]
  19. Lamb R. A., Choppin P. W. 1979; Segment 8 of the influenza virus genome is unique in coding for two polypeptides. Proceedings of the National Academy of Sciences, U.S.A 76:4908–4912
     [Google Scholar]
  20. Lamb R. A., Choppin P. W. 1983; Gene structure and replication of influenza virus. Annual Review of Biochemistry 52:467–506
     [Google Scholar]
  21. Lamb R. A., Lai C.-J. 1980; Sequence of interrupted and uninterrupted mRNAs and cloned DNA coding for the two overlapping nonstructural proteins of influenza virus. Cell 21:475–485
     [Google Scholar]
  22. Mccauley J. W., Mahy B. W. J. 1983; Structure and function of the influenza virus genome (a review). Biochemical Journal 211:281–294
     [Google Scholar]
  23. Maeno M., Shimizu K., Shikata T. 1988; Effective production of mouse monoclonal antibodies against influenza virus by repetitive intrasplenic immunization. Microbiology and Immunology 32:1245–1252
     [Google Scholar]
  24. Mahy B. W. J. 1983; Mutants of influenza virus. In Genetics of Influenza Viruses pp. 192–254 Palese P., Kingsbury D. W. Edited by Vienna: Springer-Verlag;
     [Google Scholar]
  25. Mark G. E., Taylor J. M., Broni B., Krug R. M. 1979; Nuclear accumulation of influenza viral RNA transcripts and the effects of cycloheximide, actinomycin D and α-amanitin. Journal of Virology 29:744–752
     [Google Scholar]
  26. Maxam A. M., Gilbert W. 1980; Sequencing end-labeled DNA with base-specific chemical cleavages. Methods in Enzymology 65:499–560
     [Google Scholar]
  27. Melton D. A., Krieg P. A., Rebagliati M. R., Maniatis T., Zinn K., Green M. R. 1984; Efficient in vitro synthesis of biologically active RNA and RNA hybridization probes from plasmids containing a bacteriophage SP6 promoter. Nucleic Acids Research 12:7035–7056
     [Google Scholar]
  28. Norton G. P., Tanaka T., Tobita K., Nakada S., Buonagurio D. A., Greenspan D., Krystal M., Palese P. 1987; Infectious influenza A and B virus variants with long carboxyl terminal deletions in the NSI polypeptides. Virology 156:204–213
     [Google Scholar]
  29. Robertson J. S., Robertson E., Roditi I., Almond J. W., Inglis S. C. 1983; Sequence analysis of fowl plague virus mutant M7 reveals a nonsense mutation in the NSI gene. Virology 126:391–394
     [Google Scholar]
  30. Sanger F., Nicklen S., Coulson A. R. 1977; DNA sequencing with chain-terminating inhibitors. Proceedings of the National Academy of Sciences, U.S.A 74:5463–5467
     [Google Scholar]
  31. Shimizu K., Mullinix M. G., Chanock R. M., Murphy B. R. 1982a; Temperature-sensitive mutants of influenza A/Udorn/72 (H3N2) virus. I. Isolation of temperature-sensitive mutants some of which exhibit host-dependent temperature sensitivity. Virology 117:38–44
     [Google Scholar]
  32. Shimizu K., Mullinix M. G., Chanock R. M., Murphy B. R. 1982b; Temperature-sensitive mutants of influenza A/Udorn/72 (H3N2) virus. II. Genetic analysis and demonstration of intrasegmental complementation. Virology 117:45–61
     [Google Scholar]
  33. Shimizu K., Mullinix M. G., Chanock R. M., Murphy B. R. 1983; Temperature-sensitive mutants of influenza A/Udorn/72 (H3N2) virus. III. Genetic analysis of temperature-dependent host range mutants. Virology 124:35–44
     [Google Scholar]
  34. Smith D. B., Inglis S. C. 1985; Regulated production of an influenza virus spliced mRNA mediated by virus-specific products. EMBO Journal 4:2312–2319
     [Google Scholar]
  35. Wolstenholme A. J., Barrett T., Nichol S. T., Mahy W. J. 1980; Influenza virus-specific RNA and protein synthesis in cells infected with temperature-sensitive mutants defective in the genome segment encoding non-structural proteins. Journal of Virology 35:1–7
     [Google Scholar]
  36. Young J. F., Desselberger U., Palese P., Ferguson B., Shatzman A. R., Rosenberg M. 1983; Efficient expression of influenza virus NSI non-structural proteins in E. coli. Proceedings of the National Academy of Sciences, U.S.A 80:6105–6109
     [Google Scholar]
/content/journal/jgv/10.1099/0022-1317-71-6-1283
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Analysis of influenza A virus temperature-sensitive mutants with mutations in RNA segment 8
J. Gen. Virol. 71, 1283 (1990); https://doi.org/10.1099/0022-1317-71-6-1283
/content/journal/jgv/10.1099/0022-1317-71-6-1283
/content/journal/jgv/10.1099/0022-1317-71-6-1283
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