Skip to main content
PMC home page
Journal of Virology logoLink to Journal of Virology
. 1979 Oct;32(1):187–198. doi: 10.1128/jvi.32.1.187-198.1979

Characterization of 40,000- and 25,000-dalton intermediate precursors to Rauscher murine leukemia virus gag gene products.

R B Naso, W L Karshin, Y H Wu, R B Arlinghaus
PMCID: PMC353542  PMID: 94357

Abstract

Under steady-state labeling conditions, Rauscher murine leukemia virus-infected NIH Swiss mouse cells contain at least three major polyproteins derived from the viral gag gene. They have molecular weights of 65,000, 40,000, and 25,000. They have been termed pPr65gag, Pr40gag, and pPr25gag. pPr65gag has been shown by a number of laboratories to be composed of all four core proteins (p15, pp12, p30, and p10). In this paper, Pr40gag was found to contain p30 and p10 antigenic determinants and peptide sequences, whereas pPr25gag was found to contain p15 and pp12. Pr40gag and pPr25gag are rapidly labeled precursor proteins that were detectable early in pulse-chase experiments. Both precursors disappeared during the later stages of the chase period concurrent with the appearance of the mature viral core proteins. pPr65gag and pPr25gag were found to be phosphorylated, pPr25 having a higher specific activity of 32P than pPr65. In spite of this, peptide mapping studies, as well as the identification of the phosphorylated amino acid residues of pPr65, and pPr25, and pp12, indicated that the same sites are phosphorylated regardless of whether the precursors or the mature pp12 are examined.

Full text

PDF
187

Images in this article

189Image
on p.189
189Image
on p.189
194Image
on p.194
195Image
on p.195
196Image
on p.196

Selected References

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

  1. Arcement L. J., Karshin W. L., Naso R. B., Arlinghaus R. B. "gag" polyprotein precursors of Rauscher murine leukemia virus. Virology. 1977 Sep;81(2):284–297. doi: 10.1016/0042-6822(77)90145-3. [DOI] [PubMed] [Google Scholar]
  2. Arcement L. J., Karshin W. L., Naso R. B., Jamjoom G., Arlinghaus R. B. Biosynthesis of Rauscher leukemia viral proteins: presence of p30 and envelope p15 sequences in precursor polypeptides. Virology. 1976 Feb;69(2):763–774. doi: 10.1016/0042-6822(76)90504-3. [DOI] [PubMed] [Google Scholar]
  3. Baltimore D. Tumor viruses: 1974. Cold Spring Harb Symp Quant Biol. 1975;39(Pt 2):1187–1200. doi: 10.1101/sqb.1974.039.01.137. [DOI] [PubMed] [Google Scholar]
  4. 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]
  5. 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]
  6. Brugge J., Erikson E., Collett M. S., Erikson R. I. Peptide analysis of the transformation-specific antigen from avian sarcoma virus-transformed cells. J Virol. 1978 Jun;26(3):773–782. doi: 10.1128/jvi.26.3.773-782.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Erikson E., Brugge J. S., Erikson R. L. Phosphorylated and nonphosphorylated forms of avian sarcoma virus polypeptide p19. Virology. 1977 Jul 1;80(1):177–185. doi: 10.1016/0042-6822(77)90390-7. [DOI] [PubMed] [Google Scholar]
  8. 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]
  9. Jamjoom G. A., Naso R. B., Arlinghaus R. B. Selective decrease in the rate of cleavage of an intracellular precursor to Rauscher leukemia virus p30 by treatment of infected cells with actinomycin D. J Virol. 1976 Sep;19(3):1054–1072. doi: 10.1128/jvi.19.3.1054-1072.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. 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]
  11. 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]
  12. Kopchick J. J., Karshin W. L., Arlinghaus R. B. Tryptic peptide analysis of gag and gag-pol gene products of Rauscher murine leukemia virus. J Virol. 1979 May;30(2):610–623. doi: 10.1128/jvi.30.2.610-623.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. 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]
  14. Laskey R. A., Mills A. D. Quantitative film detection of 3H and 14C in polyacrylamide gels by fluorography. Eur J Biochem. 1975 Aug 15;56(2):335–341. doi: 10.1111/j.1432-1033.1975.tb02238.x. [DOI] [PubMed] [Google Scholar]
  15. Murphy E. C., Jr, Arlinghaus R. B. Tryptic peptide analyses of polypeptides generated by premature termination of cell-free protein synthesis allow a determination of the Rauscher leukemia virus gag gene order. J Virol. 1978 Dec;28(3):929–935. doi: 10.1128/jvi.28.3.929-935.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Naso R. B., Arcement L. J., Arlinghaus R. B. Biosynthesis of Rauscher leukemia viral proteins. Cell. 1975 Jan;4(1):31–36. doi: 10.1016/0092-8674(75)90130-0. [DOI] [PubMed] [Google Scholar]
  17. 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]
  18. Pal B. K., Roy-Burman P. Phosphoproteins: structural components of oncornaviruses. J Virol. 1975 Mar;15(3):540–549. doi: 10.1128/jvi.15.3.540-549.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Pal B. K., Roy-Burman P. RNA tumor virus phosphoproteins: phosphorylation of precursor and processed polypeptides. Biochem Biophys Res Commun. 1978 Mar 30;81(2):344–350. doi: 10.1016/0006-291x(78)91539-5. [DOI] [PubMed] [Google Scholar]
  20. Sen A., Sherr C. J., Todaro G. J. Phosphorylation of murine type C viral p12 proteins regulates their extent of binding to the homologous viral RNA. Cell. 1977 Mar;10(3):489–496. doi: 10.1016/0092-8674(77)90036-8. [DOI] [PubMed] [Google Scholar]
  21. Shapiro S. Z., Strand M., August J. T. High molecular weight precursor polypeptides to structural proteins of Rauscher murine leukemia virus. J Mol Biol. 1976 Nov 15;107(4):459–477. doi: 10.1016/s0022-2836(76)80078-2. [DOI] [PubMed] [Google Scholar]
  22. 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]
  23. 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]
  24. 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]
  25. Witte O. N., Baltimore D. Relationship of retrovirus polyprotein cleavages to virion maturation studied with temperature-sensitive murine leukemia virus mutants. J Virol. 1978 Jun;26(3):750–761. doi: 10.1128/jvi.26.3.750-761.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. 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]

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

RESOURCES