Skip to main content
NCBI home page
Journal of Virology logoLink to Journal of Virology
. 1983 Mar;45(3):1177–1182. doi: 10.1128/jvi.45.3.1177-1182.1983

Detection of an 85,000-dalton phosphoprotein in ts110 murine sarcoma virus-infected cells with antiserum against a v-mos peptide.

J Papkoff, T Hunter
PMCID: PMC256530  PMID: 6300455

Abstract

We have used an antiserum directed against a synthetic v-mos peptide (anti-C3 serum) to screen ts110 murine sarcoma virus (MuSV)-infected cells for the presence of v-mos-encoded proteins. Anti-C3 serum specifically recognized an 85,000-dalton protein doublet (P85) from [35S]methionine-labeled ts110 MuSV-infected producer cells grown at 32 degrees C, the permissive temperature for transformation. The P85 doublet was also recognized by an antiserum directed against the viral gag protein p15. P85 was present but at 2- to 10-fold-lower levels in ts110 MuSV-infected producer cells grown at 39 degrees C, the restrictive temperature for transformation. The P85gag-mos fusion product was the only v-mos protein reproducibly detected in this ts110 MuSV-transformed cell line. Immunoprecipitation of 32P-labeled cells with anti-C3 serum revealed that the upper band of the P85 doublet is phosphorylated, containing mostly phosphoserine and some phosphothreonine. Cells acutely infected with ts110 MuSV contained slightly higher levels of P85 than did the ts110 MuSV-infected producer cell line. Anti-C3 serum specifically recognized a 33,000-dalton protein (p33) in the acutely infected cells labeled with [35S]methionine. p33 was present in trace amounts and may represent a previously unidentified ts110 MuSV-encoded v-mos protein.

Full text

PDF
1177

Images in this article

1178Image
on p.1178
1180Image
on p.1180
1181Image
on p.1181

Selected References

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

  1. Andersson P., Goldfarb M. P., Weinberg R. A. A defined subgenomic fragment of in vitro synthesized Moloney sarcoma virus DNA can induce cell transformation upon transfection. Cell. 1979 Jan;16(1):63–75. doi: 10.1016/0092-8674(79)90188-0. [DOI] [PubMed] [Google Scholar]
  2. Blair D. G., Hull M. A., Finch E. A. The isolation and preliminary characterization of temperature-sensitive transformation mutants of Moloney sarcoma virus. Virology. 1979 Jun;95(2):303–316. doi: 10.1016/0042-6822(79)90486-0. [DOI] [PubMed] [Google Scholar]
  3. Blair D. G., McClements W. L., Oskarsson M. K., Fischinger P. J., Vande Woude G. F. Biological activity of cloned Moloney sarcoma virus DNA: Terminally redundant sequences may enhance transformation efficiency. Proc Natl Acad Sci U S A. 1980 Jun;77(6):3504–3508. doi: 10.1073/pnas.77.6.3504. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Blair D. G., Oskarsson M., Wood T. G., McClements W. L., Fischinger P. J., Vande Woude G. G. Activation of the transforming potential of a normal cell sequence: a molecular model for oncogenesis. Science. 1981 May 22;212(4497):941–943. doi: 10.1126/science.7233190. [DOI] [PubMed] [Google Scholar]
  5. Canaani E., Robbins K. C., Aaronson S. A. The transforming gene of Moloney murine sarcoma virus. Nature. 1979 Nov 22;282(5737):378–383. doi: 10.1038/282378a0. [DOI] [PubMed] [Google Scholar]
  6. Donoghue D. J. Demonstration of biological activity and nucleotide sequence of an in vitro synthesized clone of the Moloney murine sarcoma virus mos gene. J Virol. 1982 May;42(2):538–546. doi: 10.1128/jvi.42.2.538-546.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Horn J. P., Wood T. G., Blair D. G., Arlinghaus R. B. Partial characterization of a moloney murine sarcoma virus 85,000-dalton polypeptide whose expression correlates with the transformed phenotype in cells infected with a temperature-sensitive mutant virus. Virology. 1980 Sep;105(2):516–525. doi: 10.1016/0042-6822(80)90052-5. [DOI] [PubMed] [Google Scholar]
  8. Horn J. P., Wood T. G., Murphy E. C., Jr, Blair D. G., Arlinghaus R. B. A selective temperature-sensitive defect in viral RNA expression in cells infected with a ts transformation mutant of murine sarcoma virus. Cell. 1981 Jul;25(1):37–46. doi: 10.1016/0092-8674(81)90229-4. [DOI] [PubMed] [Google Scholar]
  9. Junghans R. P., Murphy E. C., Jr, Arlinghaus R. B. Electron microscopic analysis of ts1 10 Moloney mouse sarcoma virus, a variant of wild-type virus with two RNAs containing large deletions. J Mol Biol. 1982 Oct 25;161(2):229–250. doi: 10.1016/0022-2836(82)90150-4. [DOI] [PubMed] [Google Scholar]
  10. Moloney J. B. A virus-induced rhabdomyosarcoma of mice. Natl Cancer Inst Monogr. 1966 Sep;22:139–142. [PubMed] [Google Scholar]
  11. Murphy E. C., Jr, Arlinghaus R. B. Comparative tryptic peptide analysis of candidate P85gag-mos of ts110 Moloney murine sarcoma virus and P38-P23 mos gene-related proteins of wild-type virus. Virology. 1982 Sep;121(2):372–383. doi: 10.1016/0042-6822(82)90175-1. [DOI] [PubMed] [Google Scholar]
  12. Papkoff J., Hunter T., Beemon K. In vitro translation of virion RNA from Moloney murine sarcoma virus. Virology. 1980 Feb;101(1):91–103. doi: 10.1016/0042-6822(80)90486-9. [DOI] [PubMed] [Google Scholar]
  13. Papkoff J., Lai M. H., Hunter T. Analysis of v-mos encoded proteins in cells transformed by several related murine sarcoma viruses. J Cell Biochem. 1982;19(4):349–362. doi: 10.1002/jcb.240190405. [DOI] [PubMed] [Google Scholar]
  14. Papkoff J., Lai M. H., Hunter T., Verma I. M. Analysis of transforming gene products from Moloney murine sarcoma virus. Cell. 1981 Nov;27(1 Pt 2):109–119. doi: 10.1016/0092-8674(81)90365-2. [DOI] [PubMed] [Google Scholar]
  15. Papkoff J., Verma I. M., Hunter T. Detection of a transforming gene product in cells transformed by Moloney murine sarcoma virus. Cell. 1982 Jun;29(2):417–426. doi: 10.1016/0092-8674(82)90158-1. [DOI] [PubMed] [Google Scholar]
  16. Reddy E. P., Smith M. J., Aaronson S. A. Complete nucleotide sequence and organization of the Moloney murine sarcoma virus genome. Science. 1981 Oct 23;214(4519):445–450. doi: 10.1126/science.6170110. [DOI] [PubMed] [Google Scholar]
  17. Reddy E. P., Smith M. J., Canaani E., Robbins K. C., Tronick S. R., Zain S., Aaronson S. A. Nucleotide sequence analysis of the transforming region and large terminal redundancies of Moloney murine sarcoma virus. Proc Natl Acad Sci U S A. 1980 Sep;77(9):5234–5238. doi: 10.1073/pnas.77.9.5234. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Sefton B. M., Hunter T., Raschke W. C. Evidence that the Abelson virus protein functions in vivo as a protein kinase that phosphorylates tyrosine. Proc Natl Acad Sci U S A. 1981 Mar;78(3):1552–1556. doi: 10.1073/pnas.78.3.1552. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Stanker L. H., Gallick G. E., Kloetzer W. S., Murphy E. C., Jr, Arlinghaus R. B. P85: a gag-mos polyprotein encoded by ts110 Moloney murine sarcoma virus. J Virol. 1983 Mar;45(3):1183–1189. doi: 10.1128/jvi.45.3.1183-1189.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Van Beveren C., Galleshaw J. A., Jonas V., Berns A. J., Doolittle R. F., Donoghue D. J., Verma I. M. Nucleotide sequence and formation of the transforming gene of a mouse sarcoma virus. Nature. 1981 Jan 22;289(5795):258–262. doi: 10.1038/289258a0. [DOI] [PubMed] [Google Scholar]
  21. Van Beveren C., van Straaten F., Galleshaw J. A., Verma I. M. Nucleotide sequence of the genome of a murine sarcoma virus. Cell. 1981 Nov;27(1 Pt 2):97–108. doi: 10.1016/0092-8674(81)90364-0. [DOI] [PubMed] [Google Scholar]
  22. Vande Woude G. F., Oskarsson M., McClements W. L., Enquist L. W., Blair D. G., Fischinger P. J., Maizel J. V., Sullivan M. Characterization of integrated Moloney Sarcoma proviruses and flanking host sequences cloned in bacteriophage lambda. Cold Spring Harb Symp Quant Biol. 1980;44(Pt 2):735–745. doi: 10.1101/sqb.1980.044.01.079. [DOI] [PubMed] [Google Scholar]
  23. Verma I. M., Lai M. H., Bosselman R. A., McKennett M. A., Fan H., Berns A. Molecular cloning of unintegrated Moloney mouse sarcoma virus DNA in bacteriophage lambda. Proc Natl Acad Sci U S A. 1980 Apr;77(4):1773–1777. doi: 10.1073/pnas.77.4.1773. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES