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1887

Journal of General Virology header logo Journal of General Virology

Volume 37, Issue 1

Comparative Studies of Wild-Type and ‘Cold-Mutant’ (Temperature Sensitive) Influenza Viruses: Genealogy of the Matrix (M) and Non-structural (NS) Proteins in Recombinant Cold-Adapted H3N2 Viruses No Access

Abstract

Summary

The matrix (M) protein of the H2N2 virus A/Ann Arbor/6/60 may be distinguished from M protein of several H3N2 viruses and A/New Jersey/76 (Hsw1N1) by SDS acrylamide gel electrophoresis using a discontinuous buffer system. The smallest RNA (RNA 8) of the A/Ann Arbor/6/60 virus may be distinguished from RNA 8 of several H3N2 viruses by acrylamide gel electrophoresis in 3% or 3.6% gels in the absence of urea, if electrophoresis is done at 30 to 35 °C or 20 °C respectively. Ten clones of conditionally-lethal temperature-sensitive (ts) mutants were studied, which derived their cold-adaption and ts genes from mutant A/Ann Arbor/6/60, and their haemagglutinin from the H3N2 virus A/Scotland/840/74. Each clone was found to derive its M protein from A/Ann Arbor/6/60 mutant, and its RNA 8 from A/Scotland/840/74. The only assignment of genes 7 and 8 consistent with these findings for the recombinants is that in each parent virus (and in the recombinants) gene 7 codes for M protein, and gene 8 for NS protein. Furthermore, it may be concluded from the results that the biologically important ts lesions in the A/Ann Arbor/6/60 mutant parent are not present in the NS gene. In addition to the recombinants of A/Ann Arbor/6/60 and A/Scotland/840/74, five independent ts/cold-adapted recombinants of A/Ann Arbor/6/60 mutant with H3N2 and Hsw1N1 wild-type viruses were examined, and all were found to contain the M protein of the A/Ann Arbor/6/60 mutant parent. This is suggestive that M protein may be at least partially responsible for the cold-adaptation and/or ts properties of the A/Ann Arbor/6/60 mutant and the recombinants.

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© Journal of General Virology 1977
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1977-10-01
2025-05-24
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References

  1. Beare A. S., Hall T. S. 1971; Recombinant influenza A viruses as live vaccines for man. Lancet ii:1271–1273
     [Google Scholar]
  2. Beare A. S., Schild G. C., Craig J. W. 1975; Trials in man with live recombinants made from A/PR/8/34 (H0N1) and wild H3N2 influenza virus. Lancet ii:729–732
     [Google Scholar]
  3. Floyd R. W., Stone M. P., Joklik W. J. 1974; Separation of single-stranded ribonucleic acids by acrylamide-agarose-urea gel electrophoresis. Analytical Biochemistry 59:599–609
     [Google Scholar]
  4. Gross K., Probst E., Schaffner W., Birnstiel M. 1976; Molecular analysis of the histone gene cluster of Psammeclimus miliaris: Ⅰ. Fractionation and identification of five individual histone m RNA’s. Cell 8:455–469
     [Google Scholar]
  5. Kendal A. P., Cox N. J., Spring S. B., Maassab H. F. 1977; Biochemical characteristics of recombinant viruses derived at suboptimal temperatures. In Negative Strand Viruses and the Host Cell (in the press)Edited by Mahy B. W. J., Barry R. D. London: Academic Press;
     [Google Scholar]
  6. Kendal A. P., Kiley M. P., Maassab H. R. 1973; Comparative studies of wild-type and ‘cold-mutant’ (temperature-sensitive) influenza virus: polypeptide synthesis by an Asian (H2N2) strain and its cold-adapted variant. Journal of Virology 12:1503–1511
     [Google Scholar]
  7. Kilbourne E. D., Schulman J. L., Schild G. C., Schloer G., Swanson J., Bucher D. 1971; Correlated studies of a recombinant influenza-virus vaccine. 1. Derivation and characterization of virus and vaccine. Journal of Infectious Diseases 124:449–462
     [Google Scholar]
  8. Laemmli V. K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, London 227:680–685
     [Google Scholar]
  9. Laver W. G., Downie J. C. 1976; Influenza virus recombination, 1. Matrix protein markers and segregation during mixed infection. Virology 70:105–117
     [Google Scholar]
  10. Maassab H. F. 1967; Adaptation and growth characteristics of influenza virus at 25 °C.. Nature, London 213:612–614
     [Google Scholar]
  11. Maassab H. F. 1975; Properties of influenza virus ‘cold’ recombinants. In Negative Strand Viruses vol 2 pp 755–763 Edited by Mahy B. W. J., Barry R. D. London: Academic Press;
     [Google Scholar]
  12. Maassab H. F., Kendal A. P., Davenport F. M. 1972; Hybrid formation of influenza virus at 25 °C. Proceedings of the Society for Experimental Biology and Medicine 139:768–773
     [Google Scholar]
  13. Morris C. R., Freestone D. S., Stealey V. M., Oliver P. R. 1975; Recombinant WRL 105 strain live attenuated influenza vaccine. Lancet ii:196–199
     [Google Scholar]
  14. Murphy B. R., Chalhub E. G., Nusinoff S. R., Chanock R. M. 1972; Temperature-sensitive mutants of influenza virus. Ⅱ. Attenuation of ts recombinants for man. Journal of Infectious Diseases 126:170–178
     [Google Scholar]
  15. Palese P., Ritchey M. B. 1977; Live attenuated influenza virus vaccines. Strains with temperature-sensitive defects in P3 protein and nucleoprotein. Virology 78:183–191
     [Google Scholar]
  16. Palese P., Schulman J. L. 1976a; Mapping of the influenza virus genome: identification of the hemagglutinin and the neuraminidase genes. Proceedings of the National Academy of Sciences of the United States of America 73:2142–2146
     [Google Scholar]
  17. Palese P., Schulman J. L. 1976b; RNA pattern of ‘swine’ influenza virus isolated from man is similar to those of other swine influenza viruses. Nature, London 263:528–530
     [Google Scholar]
  18. Pons M. W. 1976; A reexamination of influenza single- and double-stranded RNAs by gel electrophoresis. Virology 69:789–792
     [Google Scholar]
  19. Richman D. G., Murphy B. R., Spring S. B., Coleman M. T., Chanock R. M. 1975; Temperature-sensitive mutants of influenza virus. IX. Genetic and biological characterization of TS-1(E) lesions when transferred to a 1972 (H3N2) influenza A virus. Virology 66:551–562
     [Google Scholar]
  20. Ritchey M. B., Palese P., Schulman J. L. 1976; Mapping of the influenza virus genome. III. Identification of genes coding for nucleoprotein, membrane protein, and nonstructural protein. Journal of Virology 20:307–313
     [Google Scholar]
  21. Spring S. B., Maassab H. F., Kendal A. P., Murphy B. R., Chanock R. M. 1977a; Cold adapted variants of influenza A. I. Comparison of the genetic properties of t.s. mutants and five cold adapted variants of influenza virus A. Virology 77:337–343
     [Google Scholar]
  22. Spring S. B., Maassab H. F., Kendal A. P., Murphy B. R., Chanock R. M. 1977b; Cold adapted variants of influenza A. Ⅱ. Comparison of the genetic and biological properties of ts mutants and 4 recombinants of the cold-adapted A/AA/6/60 strain. Archives of Virology (in the press)
    [Google Scholar]
  23. Spring S. B., Nusinoff S. R., Mills J., Richman D. D., Tierney E. L., Murphy B. R., Channock R. M. 1975; Temperature-sensitive mutants of influenza virus. VI. Transfer of TS lesions from Asian subtype of influenza A virus (H2N2) to the Hong Kong subtype (H3N2). Virology 66:522–532
     [Google Scholar]
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Comparative Studies of Wild-Type and ‘Cold-Mutant’ (Temperature Sensitive) Influenza Viruses: Genealogy of the Matrix (M) and Non-structural (NS) Proteins in Recombinant Cold-Adapted H3N2 Viruses
J. Gen. Virol. 37, 145 (1977); https://doi.org/10.1099/0022-1317-37-1-145
/content/journal/jgv/10.1099/0022-1317-37-1-145
/content/journal/jgv/10.1099/0022-1317-37-1-145
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