1887

Abstract

SUMMARY

Cytoplasmic RNA synthesis can be detected in vaccinia virus-infected HeLa cells in the presence of 2 µg/ml but not 20 µg/ml of actinomycin D. When RNA synthesis is observed protein synthesis is inhibited in infected, treated cells. We had previously noted that such a correlation may also be observed in infected, cycloheximide-treated cells. If actinomycin D (20 µg/ml) is added to these cells at various times after infection and treatment, the inhibition of protein synthesis seen upon removal of cycloheximide does not continue beyond the point to which it had developed before the actinomycin D was added. These results indicate that the inhibition of protein synthesis can be correlated with the amount of cytoplasmic RNA synthesized in infected cells and that this RNA synthesis and the subsequent inhibition of protein synthesis can be prevented by sufficiently high concentrations of actinomycin D. The cytoplasmic RNA which is synthesized does not appear to consist of double-stranded RNA nor of extensive self complementary regions. The cytoplasmic RNA synthesized in infected, cycloheximide treated cells appears to consist of early virus mRNA which can function as mRNA in a cell-free system derived from normal cells. An examination of the phosphorylation of ribosomal proteins shows six additional phosphoproteins in infected cells, two of which may be observed in infected cycloheximide-treated cells, suggesting that phosphorylation of ribosomal proteins cannot be directly correlated with the inhibition of overall protein synthesis seen in infected cycloheximide-treated cells.

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1979-09-01
2024-04-19
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References

  1. Bablanian R. 1975; Structural and functional alterations in cultured cells infected with cytocidal viruses. Progress in Medical Virology 19:40–83
    [Google Scholar]
  2. Bablanian R., Eggers H. J., Tamm I. 1965; Studies on the mechanism of poliovirus-induced cell damage. I. The relation between poliovirus-induced metabolic and morphological alterations in cultured cells. Virology 26:114–212
    [Google Scholar]
  3. Bablanian R., Esteban M., Baxt B., Sonnabend J. 1978; Studies on the mechanisms of vaccinia virus cytopathic effects. I. Inhibition of protein synthesis in infected cells is associated with virus-induced RNA synthesis. Journal of General Virology 39:391–402
    [Google Scholar]
  4. Becker Y., Joklik W. K. 1964; Messenger RNA in cells infected with vaccinia virus. Proceedings of the National Academy of Sciences of the United States of America 51:577–585
    [Google Scholar]
  5. Ben-Hamida F., Beaud G. 1978; In vitro inhibition of protein synthesis by purified cores from vaccinia virus. Proceedings of the National Academy of Sciences of the United States of America 75:175–179
    [Google Scholar]
  6. Boone R., Moss B. 1978; Synthesis of polyadenylated RNA containing complementary sequences during vaccinia virus infection. Abstracts of the Annual Meeting of the American Society for Microbiology pp 248
    [Google Scholar]
  7. Bramhall S., Noack N., Wu M., Loewenberg J. R. 1969; A simple colorimetric method for determination of protein. Analytical Biochemistry 31:146–148
    [Google Scholar]
  8. Colby C., Duesberg P. H. 1969; Double-stranded RNA in vaccine virus infected cells. Nature, London 222:940–944
    [Google Scholar]
  9. Colby C., Jurale C., Kates J. R. 1971; Mechanism of synthesis of vaccinia virus double-stranded ribonucleic acid in vivo and in vitro. Journal of Virology 7:71–76
    [Google Scholar]
  10. Duesberg P. H., Colby C. 1969; On the biosynthesis and structure of double-stranded RNA in vaccinia virus-infected cells. Proceedings of the National Academy of Sciences of the United States of America 64:396–403
    [Google Scholar]
  11. Fairbanks G., Levinthal C., Reeder R. H. 1965; Analysis of 14C labelled proteins by disc electrophoresis. Biochemical and Biophysical Research Communications 20:393–396
    [Google Scholar]
  12. Farrell P. J., Balkow K., Hunt T., Jackson R. J., Trachsel H. 1977; Phosphorylation of initiation factor elF-2 and the control of reticulocyte protein synthesis. Cell n:187–200
    [Google Scholar]
  13. Holowczak J. A., Joklik W. K. 1967; Studies on the properties of vaccinia virus. I. Structural proteins of virions and cores. Virology 33:717–725
    [Google Scholar]
  14. Hunt T., Ehrenfeld E. 1971; Cytoplasm from poliovirus-infected HeLa cells inhibits cell-free haemoglobin synthesis. Nature New Biology 230:91–94
    [Google Scholar]
  15. Hunter T., Hunt T., Jackson R. J., Robertson H. D. 1975; The characteristics of inhibition of protein synthesis by double-stranded ribonucleic acid in reticulocyte lysates. Journal of Biological Chemistry 250:109–417
    [Google Scholar]
  16. Kaempfer R., Kaufman J. 1973; Inhibition of cellular protein synthesis by double-stranded RNA: inactivation of an initiation factor. Proceedings of the National Academy of Sciences of the United States of America 70:1222–1226
    [Google Scholar]
  17. Kaerlein M., Horak I. 1976; Phosphorylation of ribosomal proteins in HeLa cells infected with vaccinia virus. Nature, London 259:250–251
    [Google Scholar]
  18. Kates J., Beeson J. 1970; Ribonucleic acid synthesis in vaccinia virus. II. Synthesis of polyriboadenylic acid. Journal of Molecular Biology 50:19–33
    [Google Scholar]
  19. Kates J. R., Mcauslan B. R. 1967; Messenger RNA synthesis by a ‘coated’ viral genome. Proceedings of the National Academy of Sciences of the United States of America 57:314–320
    [Google Scholar]
  20. Kit S., Dubbs D. R. 1962; Biochemistry of vaccinia-infected mouse fibroblasts (strain L-M). I. Effects on nucleic acid and protein synthesis. Virology 18:274–285
    [Google Scholar]
  21. Laemmli U. K. 1970; Cleavage of structural proteins during assembly of the head of bacteriophage T4. Nature, London 227:680–685
    [Google Scholar]
  22. Lebleu B., Sen. G. E., Shaila S., Cabrer B., Lengyel P. 1976; Interferon, double-stranded RNA, and protein phosphorylation. Proceedings of the National Academy of Sciences of the United States of America 1:3107–3111
    [Google Scholar]
  23. Mans R. J., Novelli O. D. 1961; Measurement of the incorporation of radioactive amino acids into protein by a filter-paper disk method. Archives of Biochemistry and Biophysics 94:48–53
    [Google Scholar]
  24. Metz D. H., Esteban M. 1972; Interferon inhibits viral protein synthesis in L cells infected with vaccinia virus. Nature, London 238:385–388
    [Google Scholar]
  25. Moss B. 1968; Inhibition of HeLa cells protein synthesis by the vaccinia virion. Journal of Virology 2:1028–1037
    [Google Scholar]
  26. Oda K., Joklik W. K. 1967; Hybridization and sedimentation studies on ‘early’ and ‘fate’ vaccinia messenger RNA. Journal o Molecular Biology 27:395–419
    [Google Scholar]
  27. Person A., Beaud G. 1978; Inhibition of host protein synthesis in vaccinia virus-infected cells in the presence of cordycepin (3′-deoxyadenosine). Journal of Virology 25:11–18
    [Google Scholar]
  28. Roberts W. K., Clemens M. J., Kerr I. M. 1976; Interferon-induced inhibition of proteins synthesis in L-cell extracts: an ATP-dependent step in the activation of an inhibitor by double-stranded RNA. Proceedings of the National Academy of Sciences of the United States of America 73:3136–3140
    [Google Scholar]
  29. Robertson H. D., Mathews M. B. 1973; Double-stranded RNA as an inhibitor of protein synthesis and as a substrate for a nuclease in extracts of Krebs II ascites cells. Proceedings of the National Academy of Sciences of the United States of America 70:225–229
    [Google Scholar]
  30. Rosemond-Hornbeak H., Moss B. 1975; Inhibition of host protein synthesis by vaccinia virus: fate of cell mRNA and synthesis of small poly(A)-rich poly ribonucleotides in the presence of actinomycin D. Journal of Virology 16:34–42
    [Google Scholar]
  31. Salzman N. P., Sebrino E. D. 1967; Sequential formation of vaccinia virus proteins and viral deoxyribonucleic acid. Journal of Virology 1:16–23
    [Google Scholar]
  32. Schrom M., Bablanian R. 1979; Inhibition of protein synthesis by vaccinia virus. I. Characterization of an inhibited cell free protein synthesizing system from infected cells. Virology (in the press)
    [Google Scholar]
  33. Shatkin A. J. 1963; Actinomycin D and vaccinia virus infection of HeLa cells. Nature, London 199:357–358
    [Google Scholar]
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