Skip to main content
Journal of Virology logoLink to Journal of Virology
. 1972 Jul;10(1):73–81. doi: 10.1128/jvi.10.1.73-81.1972

Characterization of the Polypeptides Formed in Response to Encephalomyocarditis Virus Ribonucleic Acid in a Cell-Free System from Mouse Ascites Tumor Cells

Ian M Kerr 1, R E Brown 1, Dorothy R Tovell 1
PMCID: PMC356427  PMID: 4339197

Abstract

The polypeptide products synthesized at different times in a cell-free system from Krebs mouse ascites tumor cells in response to the addition of encephalomyocarditis (EMC) virus ribonucleic acid (RNA) were characterized by electrophoresis on polyacrylamide gels and fingerprint analysis of their tryptic peptides. Translation of the EMC RNA genome with time occurred in a nonrandom fashion in these systems, to yield products containing sequences characteristic of both virion capsid polypeptides and EMC-specific polypeptides present only in the infected cell. The molecular weights of the products fell in a series from 20,000 to 140,000 daltons, although occasionally traces of larger polypeptides were also observed. All of the major polypeptides appeared to arise from partial or complete translation of about 60% of the EMC RNA genome. They were not formed by cleavage of a large precursor molecule. It is suggested that they are artifacts generated by premature “termination” of nascent polypeptide chains at preferred sites.

Full text

PDF
73

Images in this article

Selected References

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

  1. Boime I., Aviv H., Leder P. Protein synthesis directed by encephalomyocarditis virus RNA. II. The in vitro synthesis of high molecular weight proteins and elements of the viral capsid. Biochem Biophys Res Commun. 1971 Nov 5;45(3):788–795. doi: 10.1016/0006-291x(71)90486-4. [DOI] [PubMed] [Google Scholar]
  2. Butterworth B. E., Hall L., Stoltzfus C. M., Rueckert R. R. Virus-specific proteins synthesized in encephalomyocarditis virus-infected HeLa cells. Proc Natl Acad Sci U S A. 1971 Dec;68(12):3083–3087. doi: 10.1073/pnas.68.12.3083. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Dobos P., Kerr I. M., Martin E. M. Synthesis of capsid and noncapsid viral proteins in response to encephalomyocarditis virus ribonucleic acid in animal cell-free systems. J Virol. 1971 Oct;8(4):491–499. doi: 10.1128/jvi.8.4.491-499.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Dunker A. K., Rueckert R. R. Observations on molecular weight determinations on polyacrylamide gel. J Biol Chem. 1969 Sep 25;244(18):5074–5080. [PubMed] [Google Scholar]
  5. Jacobson M. F., Asso J., Baltimore D. Further evidence on the formation of poliovirus proteins. J Mol Biol. 1970 May 14;49(3):657–669. doi: 10.1016/0022-2836(70)90289-5. [DOI] [PubMed] [Google Scholar]
  6. Jacobson M. F., Baltimore D. Polypeptide cleavages in the formation of poliovirus proteins. Proc Natl Acad Sci U S A. 1968 Sep;61(1):77–84. doi: 10.1073/pnas.61.1.77. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. KERR I. M., MARTIN E. M., HAMILTON M. G., WORK T. S. The initiation of virus protein synthesis in Krebs ascites tumor cells infected with EMC virus. Cold Spring Harb Symp Quant Biol. 1962;27:259–269. doi: 10.1101/sqb.1962.027.001.025. [DOI] [PubMed] [Google Scholar]
  8. Kerr I. M., Cohen N., Work T. S. Factors controlling amino acid incorporation by ribosomes from krebs II mouse ascites-tumour cells. Biochem J. 1966 Mar;98(3):826–835. doi: 10.1042/bj0980826. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Kerr I. M., Martin E. M. Virus protein synthesis in animal cell-free systems: nature of the products synthesized in resonse to ribonucleic acid of encephalomyocarditis virus. J Virol. 1971 Apr;7(4):438–447. doi: 10.1128/jvi.7.4.438-447.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Kiehn E. D., Holland J. J. Synthesis and cleavage of enterovirus polypeptides in mammalian cells. J Virol. 1970 Mar;5(3):358–367. doi: 10.1128/jvi.5.3.358-367.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Levin J. G., Friedman R. M. Analysis of arbovirus ribonucleic acid forms by polyacrylamide gel electrophoresis. J Virol. 1971 Apr;7(4):504–514. doi: 10.1128/jvi.7.4.504-514.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. MARTIN E. M., MALEC J., SVED S., WORK T. S. Studies on protein and nucleic acid metabolism in virus-infected mammalian cells. 1. Encephalomyocarditis virus in Krebs II mouse-ascites-tumour cells. Biochem J. 1961 Sep;80:585–597. doi: 10.1042/bj0800585. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Mathews M., Korner A. Mammalian cell-free protein synthesis directed by viral ribonucleic acid. Eur J Biochem. 1970 Dec;17(2):328–338. doi: 10.1111/j.1432-1033.1970.tb01170.x. [DOI] [PubMed] [Google Scholar]
  14. Roumiantzeff M., Summers D. F., Maizel J. V., Jr In vitro protein synthetic activity of membrane-bound poliovirus polyribosomes. Virology. 1971 May;44(2):249–258. doi: 10.1016/0042-6822(71)90257-1. [DOI] [PubMed] [Google Scholar]
  15. Smith A. E., Marcker K. A., Mathews M. B. Translation of RNA from encephalomyocarditis virus in a mammalian cell-free system. Nature. 1970 Jan 10;225(5228):184–187. doi: 10.1038/225184a0. [DOI] [PubMed] [Google Scholar]
  16. Smith R. E., Zweerink H. J., Joklik W. K. Polypeptide components of virions, top component and cores of reovirus type 3. Virology. 1969 Dec;39(4):791–810. doi: 10.1016/0042-6822(69)90017-8. [DOI] [PubMed] [Google Scholar]
  17. Summers D. F., Maizel J. V., Jr Determination of the gene sequence of poliovirus with pactamycin. Proc Natl Acad Sci U S A. 1971 Nov;68(11):2852–2856. doi: 10.1073/pnas.68.11.2852. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Summers D. F., Maizel J. V., Jr Evidence for large precursor proteins in poliovirus synthesis. Proc Natl Acad Sci U S A. 1968 Mar;59(3):966–971. doi: 10.1073/pnas.59.3.966. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Taber R., Rekosh D., Baltimore D. Effect of pactamycin on synthesis of poliovirus proteins: a method for genetic mapping. J Virol. 1971 Oct;8(4):395–401. doi: 10.1128/jvi.8.4.395-401.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES