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. 1975 Oct;16(4):790–796. doi: 10.1128/jvi.16.4.790-796.1975

Temperature-sensitive mutants of influenza WSN virus defective in virus-specific RNA synthesis.

R M Krug, M Ueda, P Palese
PMCID: PMC354737  PMID: 1165595

Abstract

Influenza WSN virus temperature-sensitive (ts) mutants were examined for defects in viral complementary RNA (cRNA) synthesis. The synthesis of viral cRNA was determined by hybridizing RNA from infected cells to radiolabeled virion RNA of known specific activity. Mutants in complementation groups I and III synthesized little, or no, cRNA at the nonpermissive temperature (39.5 C). When cells infected by these mutants were incubated for 5 h at the permissive temperature (33 C) and were then shifted to 39.5 C, net synthesis of cRNA ceased. This strongly suggests that mutants in these two complementation groups possess a ts defect in the transciptase complex. Mutants in group II and group V synthesize reduced amounts of cRNA at 39.5 C. In contrast to the group I and group III mutants, cRNA synthesis in cells infected by a group II or a group V mutant continues after a shift-up. This indicated that these mutants do not possess a ts transcriptase complex and that these mutants are most probably defective in some step in the amplification of cRNA synthesis. As will be discussed, the most likely defect in these mutants is in the synthesis of virion-type RNA. These results suggest that there are two influenza viral gene functions required for transcription and most likely two additional gene functions required for RNA replication.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. BARRY R. D., IVES D. R., CRUICKSHANK J. G. Participation of deoxyribonucleic acid in the multiplication of influenza virus. Nature. 1962 Jun 23;194:1139–1140. doi: 10.1038/1941139a0. [DOI] [PubMed] [Google Scholar]
  2. Bishop D. H., Obijeski J. F., Simpson R. W. Transcription of the influenza ribonucleic acid genome by a virion polymerase. I. Optimal conditions for in vitro activity of the ribonucleic acid-dependent ribonucleic acid polymerase. J Virol. 1971 Jul;8(1):66–73. doi: 10.1128/jvi.8.1.66-73.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bishop D. H., Obijeski J. F., Simpson R. W. Transcription of the influenza ribonucleic acid genome by a virion polymerase. II. Nature of the in vitro polymerase product. J Virol. 1971 Jul;8(1):74–80. doi: 10.1128/jvi.8.1.74-80.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bishop D. H., Roy P., Bean W. J., Jr, Simpson R. W. Transcription of the influenza ribonucleic acid genome by a virion polymerase. 3. Completeness of the transcription process. J Virol. 1972 Oct;10(4):689–697. doi: 10.1128/jvi.10.4.689-697.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Caliguiri L. A., Compans R. W. Analysis of the in vitro product of an RNA-dependent RNA polymerase isolated from influenza virus-infected cells. J Virol. 1974 Aug;14(2):191–197. doi: 10.1128/jvi.14.2.191-197.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Emerson S. U., Wagner R. R. Dissociation and reconstitution of the transcriptase and template activities of vesicular stomatitis B and T virions. J Virol. 1972 Aug;10(2):297–309. doi: 10.1128/jvi.10.2.297-309.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Emerson S. U., Wagner R. R. L protein requirement for in vitro RNA synthesis by vesicular stomatitis virus. J Virol. 1973 Dec;12(6):1325–1335. doi: 10.1128/jvi.12.6.1325-1335.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Etkind P. R., Krug R. M. Influenza viral messenger RNA. Virology. 1974 Nov;62(1):38–45. doi: 10.1016/0042-6822(74)90301-8. [DOI] [PubMed] [Google Scholar]
  9. Ghendon Y. Z., Markushin S. G., Marchenko A. T., Sitnikov B. S., Ginzburg V. P. Biochemical characteristics of fowl plague virus TS mutants. Virology. 1973 Oct;55(2):305–319. doi: 10.1016/0042-6822(73)90170-0. [DOI] [PubMed] [Google Scholar]
  10. Gregoriades A. Actinomycin D and influenza virus multiplication in the chick embryo fibroblast. Virology. 1970 Dec;42(4):905–916. doi: 10.1016/0042-6822(70)90339-9. [DOI] [PubMed] [Google Scholar]
  11. Kingsbury D. W., Webster R. G. Cell-free translation of influenza virus messenger RNA. Virology. 1973 Dec;56(2):654–657. doi: 10.1016/0042-6822(73)90069-x. [DOI] [PubMed] [Google Scholar]
  12. Krug R. M. Cytoplasmic and nucleoplasmic viral RNPs in influenza virus-infected MDCK cells. Virology. 1972 Oct;50(1):103–113. doi: 10.1016/0042-6822(72)90350-9. [DOI] [PubMed] [Google Scholar]
  13. Krug R. M. Influenza viral RNPs newly synthesized during the latent period of viral growth in MDCK cells. Virology. 1971 Apr;44(1):125–136. doi: 10.1016/0042-6822(71)90159-0. [DOI] [PubMed] [Google Scholar]
  14. Lesnaw J. A., Reichmann M. E. RNA synthesis by temperature-sensitive mutants of vesicular stomatitis virus, New Jersey serotype. Virology. 1975 Feb;63(2):492–504. doi: 10.1016/0042-6822(75)90322-0. [DOI] [PubMed] [Google Scholar]
  15. Lewandowski L. J., Content J., Leppla S. H. Characterization of the subunit structure of the ribonucleic acid genome of influenza virus. J Virol. 1971 Nov;8(5):701–707. doi: 10.1128/jvi.8.5.701-707.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Mackenzie J. S. Isolation of temperature-sensitive mutants and the construction of a preliminary genetic map for influenza virus. J Gen Virol. 1970 Jan;6(1):63–75. doi: 10.1099/0022-1317-6-1-63. [DOI] [PubMed] [Google Scholar]
  17. Palese P., Tobita K., Ueda M., Compans R. W. Characterization of temperature sensitive influenza virus mutants defective in neuraminidase. Virology. 1974 Oct;61(2):397–410. doi: 10.1016/0042-6822(74)90276-1. [DOI] [PubMed] [Google Scholar]
  18. Perlman S. M., Huang A. S. RNA synthesis of vesicular stomatitis virus. V. Interactions between transcription and replication. J Virol. 1973 Dec;12(6):1395–1400. doi: 10.1128/jvi.12.6.1395-1400.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Pons M. W. Studies on the replication of influenza virus RNA. Virology. 1972 Mar;47(3):823–832. doi: 10.1016/0042-6822(72)90574-0. [DOI] [PubMed] [Google Scholar]
  20. Pons M. W. The inhibition of influenza virus RNA synthesis by actinomycin D and cycloheximide. Virology. 1973 Jan;51(1):120–128. doi: 10.1016/0042-6822(73)90372-3. [DOI] [PubMed] [Google Scholar]
  21. Printz-Ané C., Combard A., Martinet C. Study of the transcription and the replication of vesicular stomatitis virus by using temperature-sensitive mutants. J Virol. 1972 Nov;10(5):889–895. doi: 10.1128/jvi.10.5.889-895.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Scholtissek C., Kruczinna R., Rott R., Klenk H. D. Characteristics of an influenza mutant temperature-sensitive for viral RNA synthesis. Virology. 1974 Apr;58(2):317–322. doi: 10.1016/0042-6822(74)90067-1. [DOI] [PubMed] [Google Scholar]
  23. Scholtissek C., Rott R. Synthesis in vivo of influenza virus plus and minus strand RNA and its preferential inhibition by antibiotics. Virology. 1970 Apr;40(4):989–996. doi: 10.1016/0042-6822(70)90145-5. [DOI] [PubMed] [Google Scholar]
  24. Simpson R. W., Hirst G. K. Temperature-sensitive mutants of influenza A virus: isolation of mutants and preliminary observations on genetic recombination and complementation. Virology. 1968 May;35(1):41–49. doi: 10.1016/0042-6822(68)90303-6. [DOI] [PubMed] [Google Scholar]
  25. Skehel J. J. Polypeptide synthesis in influenza virus-infected cells. Virology. 1972 Jul;49(1):23–36. doi: 10.1016/s0042-6822(72)80004-7. [DOI] [PubMed] [Google Scholar]
  26. Sugiura A., Tobita K., Kilbourne E. D. Isolation and preliminary characterization of temperature-sensitive mutants of influenza virus. J Virol. 1972 Oct;10(4):639–647. doi: 10.1128/jvi.10.4.639-647.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Sugiura A., Ueda M., Tobita K., Enomoto C. Further isolation and characterization of temperature-sensitive mutants of influenza virus. Virology. 1975 Jun;65(2):363–373. doi: 10.1016/0042-6822(75)90042-2. [DOI] [PubMed] [Google Scholar]
  28. Ueda M. Temperature-sensitive mutants of influenza virus. Isolation and preliminary characterization. Arch Gesamte Virusforsch. 1972;39(4):360–368. doi: 10.1007/BF01241015. [DOI] [PubMed] [Google Scholar]

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