Skip to main content
Journal of Virology logoLink to Journal of Virology
. 1975 Aug;16(2):388–396. doi: 10.1128/jvi.16.2.388-396.1975

Proteins synthesized by Semliki Forest virus and its 16 temperature-sensitive mutants.

S Keränen, L Kääriäinen
PMCID: PMC354677  PMID: 1171265

Abstract

The proteins synthesized in chicken embryo fibroblasts infected with wild-type Semliki Forest virus and 16 temperature-sensitive mutants derived from it were studied by polyacrylamide gel electrophoresis. In addition to the structural proteins, five nonvirion proteins (NVP) with molecular weight of 130,000, 97,000, 86,000, 78,000 and 62,000 were found varying amounts in cells infected with the different RNA+ mutants and also in the wild-type-infected cells. Pulse-chase experiments suggested that NVP 130, NVP 97, NVP 86, and NVP 62 are precursors presumably of the structural proteins. The amount of NVP 78 was not affected by the chase, and it may represent a translational product of the nonstructural part of the genome. The NVP 130 was shown to be a common precursor of the structural proteins by tryptic peptide mapping. Kinetic evidence from one of the mutants (ts-3) suggested that NVP 86 is one of the precursors of the capsid protein. A common feature of all the RNA+mutants was the inability to cleave the NVP 62 into E2 and E3, suggesting that this cleavage is a crucial reaction in the virus maturation.

Full text

PDF
388

Images in this article

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Burrell C. J., Martin E. M., Cooper P. D. Posttranslational cleavage of virus polypeptides in arbovirus-infected cells. J Gen Virol. 1970 Feb;6(2):319–323. doi: 10.1099/0022-1317-6-2-319. [DOI] [PubMed] [Google Scholar]
  2. Celma M. L., Ehrenfeld E. Effect of poliovirus double-stranded RNA on viral and host-cell protein synthesis. Proc Natl Acad Sci U S A. 1974 Jun;71(6):2440–2444. doi: 10.1073/pnas.71.6.2440. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Cordell-Stewart B., Taylor M. W. Effect of viral double-stranded RNA on protein synthesis in intact cells. J Virol. 1973 Feb;11(2):232–237. doi: 10.1128/jvi.11.2.232-237.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Darnall D. W., Klotz I. M. Protein subunits: a table (revised edition). Arch Biochem Biophys. 1972 Mar;149(1):1–14. doi: 10.1016/0003-9861(72)90293-7. [DOI] [PubMed] [Google Scholar]
  5. Darnbrough C., Hunt T., Jackson R. J. A complex between met-tRNA F and native 40S subunits in reticulocyte lysates and its disappearance during incubation with double-stranded RNA. Biochem Biophys Res Commun. 1972 Sep 26;48(6):1556–1564. doi: 10.1016/0006-291x(72)90891-1. [DOI] [PubMed] [Google Scholar]
  6. Ehrenfeld E., Hunt T. Double-stranded poliovirus RNA inhibits initiation of protein synthesis by reticulocyte lysates. Proc Natl Acad Sci U S A. 1971 May;68(5):1075–1078. doi: 10.1073/pnas.68.5.1075. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Fairbanks G., Steck T. L., Wallach D. F. Electrophoretic analysis of the major polypeptides of the human erythrocyte membrane. Biochemistry. 1971 Jun 22;10(13):2606–2617. doi: 10.1021/bi00789a030. [DOI] [PubMed] [Google Scholar]
  8. Friedman R. M. Structural and nonstructural proteins of an arbovirus. J Virol. 1968 Oct;2(10):1076–1080. doi: 10.1128/jvi.2.10.1076-1080.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Garoff H. Cross-linking of the spike glycoproteins in Semliki Forest virus with dimethylsuberimidate. Virology. 1974 Dec;62(2):385–392. doi: 10.1016/0042-6822(74)90400-0. [DOI] [PubMed] [Google Scholar]
  10. Garoff H., Simons K., Renkonen O. Isolation and characterization of the membrane proteins of Semliki Forest virus. Virology. 1974 Oct;61(2):493–504. doi: 10.1016/0042-6822(74)90285-2. [DOI] [PubMed] [Google Scholar]
  11. Hay A. J., Skehel J. J., Burke D. C. Proteins synthesized in chick cells following infection with Semliki Forest virus. J Gen Virol. 1968 Sep;3(2):175–184. doi: 10.1099/0022-1317-3-2-175. [DOI] [PubMed] [Google Scholar]
  12. Jones K. J., Waite M. R., Bose H. R. Cleavage of a viral envelope precursor during the morphogenesis of Sindbis virus. J Virol. 1974 Apr;13(4):809–817. doi: 10.1128/jvi.13.4.809-817.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Kaariainen L., Gomatos P. J. A kinetic analysis of the synthesis in BHK 21 cells of RNAs specific for Semliki Forest virus. J Gen Virol. 1969 Sep;5(2):251–265. doi: 10.1099/0022-1317-5-2-251. [DOI] [PubMed] [Google Scholar]
  14. Kaluza G., Schmidt M. F., Scholtissek C. Effect of 2-deoxy-D-glucose on the multiplication of Semliki Forest virus and the reversal of the block by mannose. Virology. 1973 Jul;54(1):179–189. doi: 10.1016/0042-6822(73)90127-x. [DOI] [PubMed] [Google Scholar]
  15. Käriäinen L., Simons K., von Bonsdorff C. H. Studies in subviral components of Semliki Forest virus. Ann Med Exp Biol Fenn. 1969;47(4):235–248. [PubMed] [Google Scholar]
  16. Laine R., Söderlund H., Renkonen O. Chemical composition of Semliki forest virus. Intervirology. 1973;1(2):110–118. doi: 10.1159/000148837. [DOI] [PubMed] [Google Scholar]
  17. Morser M. J., Burke D. C. Cleavage of virus-specified polypeptides in cells infected with Semliki Forest Virus. J Gen Virol. 1974 Mar;22(3):395–409. doi: 10.1099/0022-1317-22-3-395. [DOI] [PubMed] [Google Scholar]
  18. Morser M. J., Kennedy S. I., Burke D. C. Virus-specified polypeptides in cells infected with Semliki Forest virus. J Gen Virol. 1973 Oct;21:19–29. doi: 10.1099/0022-1317-21-1-19. [DOI] [PubMed] [Google Scholar]
  19. Neville D. M., Jr Molecular weight determination of protein-dodecyl sulfate complexes by gel electrophoresis in a discontinuous buffer system. J Biol Chem. 1971 Oct 25;246(20):6328–6334. [PubMed] [Google Scholar]
  20. Ranki M. Nucleocapsid and envelope protein of Semliki forest virus as affected by canavanine. J Gen Virol. 1972 Apr;15(1):59–67. doi: 10.1099/0022-1317-15-1-59. [DOI] [PubMed] [Google Scholar]
  21. Schlesinger M. J., Schlesinger S. Large-molecular-weight precursors of sindbis virus proteins. J Virol. 1973 Jun;11(6):1013–1016. doi: 10.1128/jvi.11.6.1013-1016.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Simons K., Keränen S., Käriänen L. Identification of a precursor for one of the Semliki forest virus membrane proteins. FEBS Lett. 1973 Jan 15;29(2):87–91. doi: 10.1016/0014-5793(73)80532-0. [DOI] [PubMed] [Google Scholar]
  23. Weber K., Osborn M. The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis. J Biol Chem. 1969 Aug 25;244(16):4406–4412. [PubMed] [Google Scholar]

Articles from Journal of Virology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES