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. 1979 Feb;29(2):649–656. doi: 10.1128/jvi.29.2.649-656.1979

Feline sarcoma virus-coded polyprotein: enzymatic cleavage by a type C virus-coded structural protein.

A S Khan, J R Stephenson
PMCID: PMC353197  PMID: 219252

Abstract

A purified 15,000-molecular-weight (Mr) Prague strain Rous sarcoma virus gag gene-coded structural protein, p15, was shown to enzymatically cleave the previously described 130,000 Mr feline sarcoma virus-coded polyprotein, Pr130. Cleavage products included proteins ranging in molecular weight from 12,000 to 110,000. The specificity of this cleavage reactivity was indicated by the fact that, under similar conditions, neither purified type C viral structural proteins nor nonviral proteins such as bovine serum albumin were cleaved to significant extents. Moreover, feline leukemia virus Pr65gag was efficiently cleaved, resulting in the generations of proteins of 30,000 (p30), 15,000 (p15), 12,000 (p12), and 10,000 (p10) Mr. Using enzymatically (p15) treated feline sarcoma virus Pr130 as starting material, we were able to purify a major 72,000 Mr cleavage product and to show it to contain the previously described feline sarcoma virus-coded nonstructural component.

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Selected References

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

  1. Barbacid M., Stephenson J. R., Aaronson S. A. gag Gene of mammalian type-C RNA tumour viruses. Nature. 1976 Aug 12;262(5569):554–559. doi: 10.1038/262554a0. [DOI] [PubMed] [Google Scholar]
  2. Bonner W. M., Laskey R. A. A film detection method for tritium-labelled proteins and nucleic acids in polyacrylamide gels. Eur J Biochem. 1974 Jul 1;46(1):83–88. doi: 10.1111/j.1432-1033.1974.tb03599.x. [DOI] [PubMed] [Google Scholar]
  3. Dittmar K. J., Moelling K. Biochemical properties of p15-associated protease in an avian RNA tumor virus. J Virol. 1978 Oct;28(1):106–118. doi: 10.1128/jvi.28.1.106-118.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Eisenman R., Shaikh R., Mason W. S. Identification of an avian oncovirus polyprotein in uninfected chick cells. Cell. 1978 May;14(1):89–104. doi: 10.1016/0092-8674(78)90304-5. [DOI] [PubMed] [Google Scholar]
  5. Khan A. S., Deobagkar D. N., Stephenson J. R. Isolation and characterization of a feline sarcoma virus-coded precursor polyprotein. Competition immunoassay for nonstructural components. J Biol Chem. 1978 Dec 25;253(24):8894–8901. [PubMed] [Google Scholar]
  6. Khan A. S., Stephenson J. R. Feline leukemia virus: biochemical and immunological characterization of gag gene-coded structural proteins. J Virol. 1977 Sep;23(3):599–607. doi: 10.1128/jvi.23.3.599-607.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  8. Oroszlan S., Henderson L. E., Stephenson J. R., Copeland T. D., Long C. W., Ihle J. N., Gilden R. V. Amino- and carboxyl-terminal amino acid sequences of proteins coded by gag gene of murine leukemia virus. Proc Natl Acad Sci U S A. 1978 Mar;75(3):1404–1408. doi: 10.1073/pnas.75.3.1404. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Philipson L., Andersson P., Olshevsky U., Weinberg R., Baltimore D., Gesteland R. Translation of MuLV and MSV RNAs in nuclease-treated reticulocyte extracts: enhancement of the gag-pol polypeptide with yeast suppressor tRNA. Cell. 1978 Jan;13(1):189–199. doi: 10.1016/0092-8674(78)90149-6. [DOI] [PubMed] [Google Scholar]
  10. Reynolds F. H., Jr, Hanson C. A., Stephenson J. R. Biochemical properties of avian type-C virus gag gene-coded proteins: comparison with structural proteins coded for at analogous positions within the mammalian type-C viral genome. Virology. 1978 May 1;86(1):177–185. doi: 10.1016/0042-6822(78)90018-1. [DOI] [PubMed] [Google Scholar]
  11. Reynolds F. H., Jr, Sacks T. L., Deobagkar D. N., Stephenson J. R. Cells nonproductively transformed by Abelson murine leukemia virus express a high molecular weight polyprotein containing structural and nonstructural components. Proc Natl Acad Sci U S A. 1978 Aug;75(8):3974–3978. doi: 10.1073/pnas.75.8.3974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Reynolds R. K., Stephenson J. R. Intracistronic mapping of the murine type C viral gag gene by use of conditional lethal replication mutants. Virology. 1977 Sep;81(2):328–340. doi: 10.1016/0042-6822(77)90149-0. [DOI] [PubMed] [Google Scholar]
  13. Reynolds R. K., van de Ven W. J., Stephenson J. R. Translation of type C viral RNAs in Xenopus laevis oocytes: evidence that the 120,000-molecular-weight polyprotein expressed in Abelson leukemia virus-transformed cells is virus coded. J Virol. 1978 Nov;28(2):665–670. doi: 10.1128/jvi.28.2.665-670.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Sacks T. L., Reynolds F. H., Jr, Deobagkar D. N., Stephenson J. R. Murine leukemia virus (T-8)-transformed cells: identification of a precursor polyprotein containing gag gene-coded proteins (p15 and p12) and a nonstructural component. J Virol. 1978 Sep;27(3):809–814. doi: 10.1128/jvi.27.3.809-814.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Sherr C. J., Sen A., Todaro G. J., Sliski A., Essex M. Pseudotypes of feline sarcoma virus contain an 85,000-dalton protein with feline oncornavirus-associated cell membrane antigen (FOCMA) activity. Proc Natl Acad Sci U S A. 1978 Mar;75(3):1505–1509. doi: 10.1073/pnas.75.3.1505. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Stephenson J. R., Devare S. G., Reynolds F. H., Jr Translational products of type-C RNA tumor viruses. Adv Cancer Res. 1978;27:1–53. doi: 10.1016/s0065-230x(08)60929-x. [DOI] [PubMed] [Google Scholar]
  17. Stephenson J. R., Khan A. S., Sliski A. H., Essex M. Feline oncornavirus-associated cell membrane antigen: evidence for an immunologically crossreactive feline sarcoma virus-coded protein. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5608–5612. doi: 10.1073/pnas.74.12.5608. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Van Zaane D., Dekker-Michielsen J. A., Bloemers H. P. Virus-specific precursor polypeptides in cells infected with Rauscher leukemia virus: synthesis, identification, and processing. Virology. 1976 Nov;75(1):113–129. doi: 10.1016/0042-6822(76)90011-8. [DOI] [PubMed] [Google Scholar]
  19. Vogt V. M., Eisenman R., Diggelmann H. Generation of avian myeloblastosis virus structural proteins by proteolytic cleavage of a precursor polypeptide. J Mol Biol. 1975 Aug 15;96(3):471–493. doi: 10.1016/0022-2836(75)90174-6. [DOI] [PubMed] [Google Scholar]
  20. Witte O. N., Rosenberg N., Paskind M., Shields A., Baltimore D. Identification of an Abelson murine leukemia virus-encoded protein present in transformed fibroblast and lymphoid cells. Proc Natl Acad Sci U S A. 1978 May;75(5):2488–2492. doi: 10.1073/pnas.75.5.2488. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. von der Helm K. Cleavage of Rous sarcoma viral polypeptide precursor into internal structural proteins in vitro involves viral protein p15. Proc Natl Acad Sci U S A. 1977 Mar;74(3):911–915. doi: 10.1073/pnas.74.3.911. [DOI] [PMC free article] [PubMed] [Google Scholar]

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