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. 1981 Sep;39(3):963–967. doi: 10.1128/jvi.39.3.963-967.1981

In vitro proteolytic cleavage of Gazdar murine sarcoma virus p65gag.

S Maxwell, R B Arlinghaus
PMCID: PMC171332  PMID: 6169847

Abstract

Moloney murine leukemia virus, disrupted in concentrations of 0.1 to 0.5% Nonidet P-40, catalyzed the cleavage of p65, the gag gene polyprotein of the Gazdar strain of murine sarcoma virus, into polypeptides with sizes and antigenic determinants of murine leukemia virus-specified p30, p15, pp12, and p10. Cleavage performed in the presence of 0.15% Nonidet P-40 in water yielded polypeptides of approximately 40,000 (P40) and 25,000 (P25) Mr. In vitro cleavage performed in a buffered solution containing dithiothreitol in addition to 0.1% Nonidet P-40 allowed the efficient processing of P40 to p30 and a band migrating with p10. Immunoprecipitation with monospecific sera indicated that P40 contained p30 and p10, whereas P25 contained p15 and pp12 determinants. P40 and P25 are similar in size and antigenic properties to Pr40gag and Pr25gag observed in infected cells (Naso et al, J. Virol. 32:187-198, 1979).

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Selected References

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

  1. Gazdar A. F., Phillips L. A., Sarma P. S., Peebles P. T., Chopra H. C. Presence of sarcoma genome in a "non-infectious" mammalian virus. Nat New Biol. 1971 Nov 17;234(46):69–72. doi: 10.1038/newbio234069a0. [DOI] [PubMed] [Google Scholar]
  2. Jamjoom G. A., Naso R. B., Arlinghaus R. B. Further characterization of intracellular precursor polyproteins of Rauscher leukemia virus. Virology. 1977 May 1;78(1):11–34. doi: 10.1016/0042-6822(77)90075-7. [DOI] [PubMed] [Google Scholar]
  3. Kessler S. W. Rapid isolation of antigens from cells with a staphylococcal protein A-antibody adsorbent: parameters of the interaction of antibody-antigen complexes with protein A. J Immunol. 1975 Dec;115(6):1617–1624. [PubMed] [Google Scholar]
  4. 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]
  5. Moelling K., Scott A., Dittmar K. E., Owada M. Effect of p15-associated protease from an avian RNA tumor virus on avian virus-specific polyprotein precursors. J Virol. 1980 Feb;33(2):680–688. doi: 10.1128/jvi.33.2.680-688.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Naso R. B., Karshin W. L., Wu Y. H., Arlinghaus R. B. Characterization of 40,000- and 25,000-dalton intermediate precursors to Rauscher murine leukemia virus gag gene products. J Virol. 1979 Oct;32(1):187–198. doi: 10.1128/jvi.32.1.187-198.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Ng V. L., Kopchick J. J., Karshin W. L., Wood T. G., Arlinghaus R. B. The structural relatedness of the virus core proteins of Rauscher and Moloney murine leukaemia virus. J Gen Virol. 1980 Mar;47(1):161–170. doi: 10.1099/0022-1317-47-1-161. [DOI] [PubMed] [Google Scholar]
  8. Pinter A., deHarven E. Protein composition of a defective murine sarcoma virus particle possessing the enveloped type-A morphology. Virology. 1979 Nov;99(1):103–110. doi: 10.1016/0042-6822(79)90041-2. [DOI] [PubMed] [Google Scholar]
  9. Syrewicz J. J., Naso R. B., Wang C. S., Arlinghaus R. B. Purification of large amounts of murine ribonucleic acid tumor viruses produced in roller bottle cultures. Appl Microbiol. 1972 Sep;24(3):488–494. doi: 10.1128/am.24.3.488-494.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Wood T. G., Lyons D. D., Ng V. L., Murphy E. C., Jr, Arlinghaus R. B. Characterization of viral polyproteins in cells transformed and producing Moloney murine sarcoma virus-124. Biochim Biophys Acta. 1980 Jul 29;608(2):215–231. doi: 10.1016/0005-2787(80)90168-9. [DOI] [PubMed] [Google Scholar]
  11. Yoshinaka Y., Luftig R. B. Murine leukemia virus morphogenesis: cleavage of P70 in vitro can be accompanied by a shift from a concentrically coiled internal strand ("immature") to a collapsed ("mature") form of the virus core. Proc Natl Acad Sci U S A. 1977 Aug;74(8):3446–3450. doi: 10.1073/pnas.74.8.3446. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Yoshinaka Y., Luftig R. B. Partial characterization of a P70 proteolytic factor that is present in purified virions of Rauscher leukemia virus (RLV). Biochem Biophys Res Commun. 1977 May 9;76(1):54–63. doi: 10.1016/0006-291x(77)91667-9. [DOI] [PubMed] [Google Scholar]
  13. Yoshinaka Y., Luftig R. B. Physicochemical characterization and specificity of the murine leukaemia virus Pr65gag proteolytic factor. J Gen Virol. 1980 Jun;48(Pt 2):329–340. doi: 10.1099/0022-1317-48-2-329. [DOI] [PubMed] [Google Scholar]
  14. Yoshinaka Y., Luftig R. B. Properties of a P70 proteolytic factor of murine leukemia viruses. Cell. 1977 Nov;12(3):709–719. doi: 10.1016/0092-8674(77)90271-9. [DOI] [PubMed] [Google Scholar]
  15. 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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