1887

Abstract

SUMMARY

When Semliki Forest virus ts-4 mutant infected cultures are grown at the permissive temperature (28 °C) and shifted to the restrictive temperature (39 °C), two different defects in RNA synthesis are manifested: (i) the synthesis of 26S RNA is stopped within 60 min (Saraste ., 1977), and (ii) the increase in RNA synthesizing activity ceases, in contrast to cultures maintained at 28 °C, indicating that no new active RNA polymerase is formed at 39 °C. Accumulation of a non-structural precursor protein with an apparent mol. wt. of about 220000 (ns220) was demonstrated in ts-4 infected cultures shifted to 39 °C. Ns220 was labelled during short pulses given immediately after release of protein synthesis from hypertonic initiation block, suggesting that genes coding for ns220 are located near the initiation site at the 5′-end of the 42S RNA. The viral specificity of ns220 was shown by its disappearance after a shift to 28 °C and by labelling in the presence of sucrose, when no host cell protein synthesis is detectable. The two functional defects can be explained if the polypeptides responsible for the RNA polymerizing activity and that responsible for the synthesis of 26S RNA are components of the same non-structural polyprotein. A mutation in the latter polypeptide which prevents cleavage of the polyprotein would thereby prevent the further formation of active RNA polymerase. If cleavage of the polyprotein has taken place at the permissive temperature, the RNA polymerase would remain active also at 39 °C, whereas the polypeptide responsible for 26S RNA synthesis would become inactive due to the mutation.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-39-3-463
1978-06-01
2021-10-27
Loading full text...

Full text loading...

/deliver/fulltext/jgv/39/3/JV0390030463.html?itemId=/content/journal/jgv/10.1099/0022-1317-39-3-463&mimeType=html&fmt=ahah

References

  1. Bonner M. W., Laskey R. 1974; A film detection method for tritium-labeled proteins and nucleic acids in polyacrylamide gels. European Journal of Biochemistry 46:83–88
    [Google Scholar]
  2. Bracha M., Leone A., Schlesinger M. J. 1976; Formation of a Sindbis virus nonstructural protein and its relation to 42S mRNA function. Journal of Virology 20:612–620
    [Google Scholar]
  3. Brzeski H., Kennedy S. I. T. 1977; Synthesis of Sindbis virus nonstructural polypeptides in chicken embryo fibroblasts. Journal of Virology 22:420–429
    [Google Scholar]
  4. Burge B. W., Pfefferkorn E. R. 1966; Complementation between temperature-sensitive mutants of Sindbis virus. Virology 30:214–223
    [Google Scholar]
  5. Cancedda R., Villa-Komaroff L., Lodish H., Schlesinger M. 1975; Initiation sites for translation of Sindbis virus 42S and 26S messenger RNAs. Cell 6:215–222
    [Google Scholar]
  6. Clegg J. C. S. 1975; Sequential translation of capsid and membrane proteins in alphaviruses. Nature, London 254:454–455
    [Google Scholar]
  7. Clegg J. C. S., Brzeski H., Kennedy S. I. T. 1976; RNA polymerase components in Semliki Forest virus infected cells: synthesis from large precursors. Journal of General Virology 32:413–430
    [Google Scholar]
  8. Clewley J. P., Kennedy S. I. T. 1976; Purification and polypeptide composition of Semliki Forest virus RNA polymerase. Journal of General Virology 32:395–411
    [Google Scholar]
  9. Glanville N., Lachmi B. 1977; Translation of proteins accounting for the full coding capacity of the Semliki Forest virus 42S RNA genome. Federation of the European Biochemical Societies Letters 81:399–402
    [Google Scholar]
  10. Glanville N., Lachmi B., Smith A. E., Kaariainen L. 1978; Tryptic peptide mapping of the nonstructural proteins of Semliki Forest virus. Biochimica et Biophysica Acta (in the press)
    [Google Scholar]
  11. Glanville N., Ranki M., Morser J., Kaariainen L., Smith A. E. 1976; Initiation of translation directed by 42S and 26S RNAs from Semliki Forest virus in vitro. Proceedings of the National Academy of Sciences of the United States of America 73:3059–3063
    [Google Scholar]
  12. Kaariainen L., Simons K., Von Bonsdorff C.-H. 1969; Studies on subviral components of Semliki Forest virus. Annales Medicinae Experimentalis et Biologiae Fenniae 47:235–248
    [Google Scholar]
  13. Kennedy S. I. T. 1976; Sequence relationships between the genome and the intracellular RNA species of standard and defective interfering Semliki Forest virus. Journal of Molecular Biology 108:491–511
    [Google Scholar]
  14. Keranen S., Kaariainen L. 1974; Isolation and basic characterization of temperature-sensitive mutants from Semliki Forest virus. Acta Pathologica et Microbiologica Scandinavica B82:810–820
    [Google Scholar]
  15. Keranen S., Kaariainen L. 1975; Proteins synthesized by Semliki Forest virus and its 16 temperature-sensitive mutants. Journal of Virology 16:388–396
    [Google Scholar]
  16. Lachmi B., Glanville N., Keranen S., Kaariainen L. 1975; Tryptic peptide analysis of nonstructural and structural precursor proteins from Semliki Forest virus mutant-infected cells. Journal of Virology 16:1615–1629
    [Google Scholar]
  17. Lachmi B., Kaariainen L. 1976; Sequential translation of nonstructural proteins in cells infected with Semliki Forest virus mutant. Proceedings of the National Academy of Sciences of the United States of America 73:1936–1940
    [Google Scholar]
  18. Lachmi B., Kaariainen L. 1977; Control of protein synthesis in Semliki Forest virus infected cells. Journal of Virology 22:142–149
    [Google Scholar]
  19. Neville D. M. 1971; Molecular weight determination of protein dodecyl sulfate complexes by gel electrophoresis in a discontinuous buffer. Journal of Biological Chemistry 246:6328–6334
    [Google Scholar]
  20. Nuss D. L., Oppermen H., Koch G. 1975; Selective blockage of initiation of host cell protein synthesis in RNA-virus-infected cells. Proceedings of the National Academy of Sciences of the United States of America 72:1258–1262
    [Google Scholar]
  21. Saborio J. L., Pong S. S., Kochi G. 1974; Selective and reversible inhibition of initiation of protein synthesis in mammalian cells. Journal of Molecular Biology 85:195–212
    [Google Scholar]
  22. Saraste J., Kaariainen L., Soderlund H., Keranen S. 1977; RNA synthesis directed by a temperature-sensitive mutant of Semliki Forest virus. Journal of General Virology 37:399–406
    [Google Scholar]
  23. Sawicki D. L., Kaariainen L., Lambeck C., Gomatos P. J. 1978; Mechanism for control of synthesis of Semliki Forest virus 26S and 42S RNA. Journal of Virology 25:19–27
    [Google Scholar]
  24. Scheele C. M., Peefferkorn 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]
  25. Schlesinger M. J., Schlesinger S. 1973; Large-molecular-weight precursors of Sindbis virus proteins. Journal of Virology 11:1013–1016
    [Google Scholar]
  26. Strauss J. H., Strauss E. 1977; Togaviruses. In The Molecular Biology of Animal Viruses pp 111–166 Edited by Nayak D. P. New York: Marcel Dekker;
    [Google Scholar]
  27. Wengler G., Wengler G. 1975; Studies on the synthesis of viral RNA-polymerase-template complexes in BHK 21 cells infected with Semliki Forest virus. Virology 66:322–326
    [Google Scholar]
  28. Wengler G., Wengler G. 1976; Localization of the 26S RNA sequence on the viral genome type 42S RNA isolated from Semliki Forest virus infected cells. Virology 73:190–199
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-39-3-463
Loading
/content/journal/jgv/10.1099/0022-1317-39-3-463
Loading

Data & Media loading...

Most cited this month Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error