1887

Abstract

SUMMARY

Using polyacrylamide gel electrophoresis, column chromatography on CF11 cellulose and salt precipitation, together with ribonuclease treatment, we have identified, in chick cells infected with Semliki Forest virus, four species of virus-specified single-stranded RNA, three species of double-stranded RNA and a multi-stranded RNA species. The mol. wt. of the four single-stranded RNA species were 4.0 × 10, 3.1 × 10, 2.3 × 10 and 1.8 × 10.

The kinetics of virus-specified RNA synthesis were studied in virus-infected cells, both early in infection and when virus-specified RNA synthesis was at the maximum rate. The first species of virus RNA detected were the multi-stranded and double-stranded species of RNA. The first single-stranded species detected was the 42S virus particle RNA, at 1.5 h after infection. The average time taken to synthesize a molecule of virus-specified RNA was between 1 and 1.5 min. Multi-stranded and double-stranded RNA both had the properties expected of an intermediate in RNA synthesis, and the number of single-stranded equivalents in the multi-stranded species was 4.3. The data show that synthesis of virus RNA is mainly semi-conservative and that the virus messenger RNA is of the same polarity as the genome.

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/content/journal/jgv/10.1099/0022-1317-24-1-45
1974-07-01
2022-08-19
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References

  1. Arif B. M., Faulkner P. 1972; Genome of Sindbis virus. Journal of Virology 9:102–109
    [Google Scholar]
  2. Baltimore D. 1969; The replication of picornaviruses. In The Biochemistry of Viruses101–176 Edited by Levy H. B. New York: Marcel Dekker;
    [Google Scholar]
  3. Baltimore D. 1971; Expression of animal virus genomes. Bacteriological Reviews 35:235–241
    [Google Scholar]
  4. Baltimore D., Girard M. 1966; An intermediate in the synthesis of poliovirus RNA. Proceedings of the National Academy of Sciences of the United States of America 56:741–748
    [Google Scholar]
  5. Bishop D. H. L., Claybrook J. R., Spiegelman S. 1967; Electrophoretic separation of viral nucleic acids on polyacrylamide gels. Journal of Molecular Biology 26:373–387
    [Google Scholar]
  6. Bishop J. M., Koch G. 1969; Infectious replicative intermediate of poliovirus: purification and characterisation. Virology 37:521–534
    [Google Scholar]
  7. Cartwright K. L., Burke D. C. 1970; Virus nucleic acids formed in chick embryo cells infected with Semliki Forest virus. Journal of General Virology 6:231–248
    [Google Scholar]
  8. Dobos P., Faulkner P. 1969; Properties of 42 S and 26 S Sindbis viral ribonucleic acid species. Journal of Virology 4:429–438
    [Google Scholar]
  9. Dobos P., Faulkner P. 1970; Molecular weight of Sindbis virus ribonucleic acid as measured by polyacrylamide gel electrophoresis. Journal of Virology 6:145–147
    [Google Scholar]
  10. Franklin R. M. 1966; Purification and properties of the replicative intermediate of the RNA bacteriophage Rt7. Proceedings of the National Academy of Sciences of the United States of America 55:1504–1511
    [Google Scholar]
  11. Friedman R. M., Levy H. B., Carter W. B. 1966; Replication of Semliki Forest virus: three forms of viral RNA produced during infection. Proceedings of the National Academy of Sciences of the United States of America 56:440–446
    [Google Scholar]
  12. Friedman R. M. 1968; Replicative intermediate of an arbovirus. Journal of Virology 2:547–552
    [Google Scholar]
  13. Iglewski W. J., Franklin R. M. 1967; Denaturation and renaturation of viral ribonucleic acid i. Annealing R17 ribonucleic acid with denatured replicative form or with denatured replicative intermediate. Journal of Virology 1:793–803
    [Google Scholar]
  14. Kääriäinen L., Gomatos P. J. 1969; A kinetic analysis of the synthesis in BHK21 cells of RNAs specific for Semliki Forest virus. Journal of General Virology 5:251–265
    [Google Scholar]
  15. Kennedy S. I. T. 1972; Isolation and identification of the virus-specified RNA species found on membrane-bound polyribosomes of chick embryo cells infected with Semliki Forest virus. Biochemical and Biophysical Research Communications 48:1254–1258
    [Google Scholar]
  16. Kennedy S. I. T., Burke D. C. 1972; Studies on the structural proteins of Semliki Forest virus. Journal of General Virology 14:87–98
    [Google Scholar]
  17. Levin J. G., Friedman R. M. 1971; Analysis of arbovirus ribonucleic acid forms by polyacrylamide gel electrophoresis. Journal of Virology 7:504–514
    [Google Scholar]
  18. Loening U. E. 1967; The fractionation of high-molecular-weight ribonucleic acid by polyacrylamide-gel electrophoresis. Biochemical Journal 102:251–257
    [Google Scholar]
  19. Loening U. E. 1968; Molecular weights of ribosomal RNA in relation to evolution. Journal of Molecular Biology 38:355–365
    [Google Scholar]
  20. Martin E. M. 1967; Mechanisms of viral RNA replication. In Genetic elements: Properties and Function117–156 Edited By Shugar D. London and New York: Academic Press;
    [Google Scholar]
  21. Michel M. R., Gomatos P. J. 1973; Semliki Forest virus – specific RNAs synthesised in vitro by enzyme from infected BHK cells. Journal of Virology n:900–914
    [Google Scholar]
  22. Montagnier L., Sanders F. K. 1963; Replicative form of encephalomyocarditis virus ribonucleic acid. Nature, London 199:664–667
    [Google Scholar]
  23. Morser M. J., Kennedy S. I. T., Burke D. C. 1973; Virus-specified polypeptides in cells infected with Semliki Forest virus. Journal of General Virology 21:19–29
    [Google Scholar]
  24. Mowshowitz D. 1973; Identification of polysomal RNA in BHK cells infected by Sindbis virus. Journal of Virology n:535–543
    [Google Scholar]
  25. Noble J., Levintow L. 1970; Dynamics of poliovirus-specific RNA synthesis and effects of inhibitors of virus replication. Virology 40:634–642
    [Google Scholar]
  26. Öberg B., Philipson L. 1971; Replicative structures of poliovirus RNA in vivo . Journal of Molecular Biology 58:725–737
    [Google Scholar]
  27. Scheele C. M., Pfefferkorn E. R. 1970; Virus-specific proteins synthesized in cells infected with RNA+ temperature-sensitive mutants of Sindbis virus. Journal of Virology 5:329–337
    [Google Scholar]
  28. Simmons D. T., Strauss J. H. 1972a; Replication of Sindbis virus. I. Relative size and genetic content of 26S and 49S RNA. Journal of Molecular Biology 71:599–613
    [Google Scholar]
  29. Simmons D. T., Strauss J. H. 1972b; Replication of Sindbis virus. II. Multiple forms of double-stranded RNA isolated from infected cells. Journal of Molecular Biology 71:615–631
    [Google Scholar]
  30. Skehel J. J., Hay A. J., Burke D. C., Cartwright L. N. 1967; Effects of actinomycin D and 2-mercapto-1-(β-4-pyridethyl) benzimidazole on the incorporation of [3H]-uridine hy chick embryo cells. Biochimica et biophysica acta 142:430–439
    [Google Scholar]
  31. Sonnabend J. A., Martin E. M., Mecs E. 1967; Viral specific RNAs in infected cells. Nature, London 213:365–367
    [Google Scholar]
  32. Sreevalsan T., Lockart R. Z. Jun., Dobson M. L. Jun, Hartman K. A. 1968; Replication of Western equine encephalomyelitis virus. I. Some chemical and physical characteristics of viral ribonucleic acid. Journal of Virology 2:558–566
    [Google Scholar]
  33. Stollar V., Shenk T. E., Stollar B. D. 1972; Double-stranded RNA in hamster, chick and mosquito cells infected with Sindbis virus. Virology 47:122–132
    [Google Scholar]
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