1887

Abstract

Summary

Coronavirus JHM contained six major proteins, four of which were glycosylated. The proteins were gp170, gp98, gp65, p60, gp25 and p23. Sac(-) cells infected with JHM shut off host cell protein synthesis, and the synthesis of three major (150K, 60K and 23K) and three minor (65K, 30K and 14K) polypeptides was detected by pulse-labelling with S-methionine. Antiserum directed against purified virus proteins specifically immunoprecipitated the three major intracellular species and also the 65K minor species. During a chase period, species 150K and 23K were processed and three new immunoprecipitable species, 170K, 98K and 25K appeared. The intracellular species 170K, 98K, 65K, 60K, 25K and 23K co-electrophoresed with virion proteins.

Two-dimensional gel electrophoresis of infected cell polypeptides showed that the 60K, 23K, 25K and 14K species were relatively basic polypeptides whilst the 98K and 170K were relatively acidic and heterogeneously charged polypeptides. Additionally, a charge-size modification of the 23K species during processing was detected, which was not apparent using one-dimensional gel analysis.

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1981-03-01
2022-09-25
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References

  1. Anderson R., Cheley S., Haworth-Hatherell H. 1979; Comparison of polypeptides of two strains of murine hepatitis virus. Virology 97:492–494
    [Google Scholar]
  2. Bond C. W., Leibowitz J. L., Robb J. A. 1979; Pathogenetic murine coronaviruses. II. Characterization of virus specific proteins of murine coronaviruses JHMV and A59V. Virology 94:371–384
    [Google Scholar]
  3. Kessler S. W. 1975; Rapid isolation of antigens from cells with a staphylococcal protein A antibody absorbent: parameters of the interaction of antibody-antigen complexes with protein A. Journal of Immunology 115:1617–1624
    [Google Scholar]
  4. Laemmli U. K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, London 227:680–685
    [Google Scholar]
  5. Lai M. M. C., Stohlman S. A. 1978; RNA of mouse hepatitis virus. Journal of Virology 26:236–248
    [Google Scholar]
  6. Laskey R. A., Mills A. D. 1975; Quantitative film detection of 3H and 14C in polyacrylamide gels by fluorography. European Journal of Biochemistry 56:335–341
    [Google Scholar]
  7. Mcintosh K. 1974; Coronaviruses: a comparative review. Current Topics in Microbiology and Immunology 63:85–129
    [Google Scholar]
  8. Nagashima K., Wege H., Meyermann R., Ter Meulen V. 1978; Coronavirus induced subacute demyelinating encephalitis in rats: a morphological analysis. Acta Neuropathologica 44:63–70
    [Google Scholar]
  9. Nagashima K., Wege H., Meyermann R., Ter Meulen V. 1979; Demyelinating encephalomyelitis induced by a long term coronavirus infection in rats. Acta Neuropathologica 45:205–213
    [Google Scholar]
  10. O’farrell P. H. 1975; High resolution two-dimensional gel electrophoresis of proteins. Journal of Biological Chemistry 250:4007–4021
    [Google Scholar]
  11. O’farrell P. Z., Goodman H. M., O’farrell P. H. 1977; High resolution two dimensional electrophoresis of basic as well as acidic proteins. Cell 12:1133–1142
    [Google Scholar]
  12. Reed L. J., Muench H. 1938; A simple method of estimating fifty per cent endpoints. American Journal of Hygiene 27:493–497
    [Google Scholar]
  13. Robb J. A., Bond C. W. 1979; Coronaviridae. In Comprehensive Virology vol 14 pp 195–247 Edited by Fraenkel-Conrat H., Wagner R. R. New York: Plenum Press;
    [Google Scholar]
  14. Siddell S. G., Wege H., Barthel A., Ter Meulen V. 1980; Coronavirus JHM: cell-free synthesis of structural protein p60. Journal of Virology 33:10–17
    [Google Scholar]
  15. Smith A. E., Wheeler T., Glanville N., Kaariainen L. 1974; Translation of Semliki Forest virus 42S RNA in a mouse cell-free system to give virus coat proteins. European Journal of Biochemistry 49:101–110
    [Google Scholar]
  16. Sturman L. S. 1977; Characterization of a coronavirus. I. Structural proteins: effect of preparative conditions on the migration of protein in polyacrylamide gels. Virology 77:637–649
    [Google Scholar]
  17. Sturman L. S., Holmes K. V. 1977; Characterization of a coronavirus. II. Glycoproteins of the viral envelope: tryptic peptide analysis. Virology 77:650–660
    [Google Scholar]
  18. Tyrrell D. A. J., Alexander D. J., Almeida J. D., Cunningham B. C., Easterday C. H., Garwes D. J., Hierholzer J. C., Kapikian M. R., Macnaughton A., Mcintosh K. 1978; Coronaviridae: second report. Intervirology 10:321–328
    [Google Scholar]
  19. Wege H., Műller A., Ter Meulen V. 1978; Genomic RNA of the murine coronavirus JHM. Journal of General Virology 41:217–227
    [Google Scholar]
  20. Wege H., Wege H., Nagashima K., Ter Meulen V. 1979; Structural polypeptides of the murine coronavirus JHM. Journal of General Virology 42:37–47
    [Google Scholar]
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