Skip to main content
PMC home page
Journal of Virology logoLink to Journal of Virology
. 1977 Sep;23(3):787–798. doi: 10.1128/jvi.23.3.787-798.1977

Common Precursor for Rauscher Leukemia Virus gp69/71, p15(E), and p12(E)

W L Karshin 1, L J Arcement 1, R B Naso 1, R B Arlinghaus 1
PMCID: PMC515890  PMID: 894795

Abstract

Rauscher murine leukemia virus glycoprotein gp69/71 and non-glycosylated p15(E) are synthesized by way of a 90,000-dalton precursor glycoprotein, termed Pr2a+b. Peptide mapping experiments showed that Pr2a+b contains all the tyrosine-containing tryptic peptides of gp69/71. Two additional tyrosine-containing tryptic peptides in Pr2a+b that are not detected in gp69/71 are found in p15(E). Thus, gp69/71 and p15(E) peptide sequences account for all the tyrosine tryptic peptides of Pr2a+b. The gene order of the two proteins was determined by pulse-labeling infected cells in the presence and absence of pactamycin at concentrations of the inhibitor that prevent initiation of translation, but not elongation. The gene order was found to be: 2HN-gp69/71-p15(E)-COOH. A newly identified major viral protein, termed p12(E), migrates in sodium dodecyl sulfate-polyacrylamide gels in the “p12” region. It is related to p15(E) as determined by tryptic mapping experiments. p15(E) and p12(E) are not phosphorylated, and both can be separated from phosphoprotein p12 by guanidine hydrochloride-agarose chromatography. p12(E) and p15(E) elute in the void volume fraction, whereas phosphoprotein p12 elutes between p15 and p10. The two p12 proteins can also be separated from each other by two-dimensional gel electrophoresis involving isoelectric focusing in the first dimension and sodium dodecyl sulfate-gel electrophoresis in the second dimension.

Full text

PDF
787

Images in this article

789Image
on p.789
790Image
on p.790
791Image
on p.791
794Image
on p.794
795Image
on p.795

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. 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]
  5. Butterworth B. E., Rueckert R. R. Gene order of encephalomyocarditis virus as determined by studies with pactamycin. J Virol. 1972 May;9(5):823–828. doi: 10.1128/jvi.9.5.823-828.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Duesberg P. H., Martin G. S., Vogt P. K. Glycoprotein components of avian and murine RNA tumor viruses. Virology. 1970 Aug;41(4):631–646. doi: 10.1016/0042-6822(70)90428-9. [DOI] [PubMed] [Google Scholar]
  7. Famulari N. G., Buchhagen D. L., Klenk H. D., Fleissner E. Presence of murine leukemia virus envelope proteins gp70 and p15(E) in a common polyprotein of infected cells. J Virol. 1976 Nov;20(2):501–508. doi: 10.1128/jvi.20.2.501-508.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Ikeda H., Hardy W., Jr, Tress E., Fleissner E. Chromatographic separation and antigenic analysis of proteins of the oncornaviruses. V. Identification of a new murine viral protein, p15(E). J Virol. 1975 Jul;16(1):53–61. doi: 10.1128/jvi.16.1.53-61.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. 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]
  10. 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]
  11. Jamjoom G., Karshin W. L., Naso R. B., Arcement L. J., Arlinghaus R. B. Proteins of Rauscher murine leukemia virus: resolution of a 70,000-dalton, Nonglycosylated polypeptide containing p30 peptide sequences. Virology. 1975 Nov;68(1):135–145. doi: 10.1016/0042-6822(75)90155-5. [DOI] [PubMed] [Google Scholar]
  12. Klenk H. D., Scholtissek C., Rott R. Inhibition of glycoprotein biosynthesis of influenza virus by D-glucosamine and 2-deoxy-D-glucose. Virology. 1972 Sep;49(3):723–734. doi: 10.1016/0042-6822(72)90529-6. [DOI] [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. Laver W. G. Separation of two polypeptide chains from the hemagglutinin subunit of influenza virus. Virology. 1971 Jul;45(1):275–288. doi: 10.1016/0042-6822(71)90134-6. [DOI] [PubMed] [Google Scholar]
  16. Lazarowitz S. G., Compans R. W., Choppin P. W. Influenza virus structural and nonstructural proteins in infected cells and their plasma membranes. Virology. 1971 Dec;46(3):830–843. doi: 10.1016/0042-6822(71)90084-5. [DOI] [PubMed] [Google Scholar]
  17. McLellan W. L., August J. T. Analysis of the envelope of Rauscher murine oncornavirus: in vitro labeling of glycopeptides. J Virol. 1976 Dec;20(3):627–636. doi: 10.1128/jvi.20.3.627-636.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Moroni C. Structural proteins of Rauscher leukemia virus and Harvey sarcoma virus. Virology. 1972 Jan;47(1):1–7. doi: 10.1016/0042-6822(72)90232-2. [DOI] [PubMed] [Google Scholar]
  19. 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]
  20. Naso R. B., Arcement L. J., Karshin W. L., Jamjoom G. A., Arlinghaus R. B. A fucose-deficient glycoprotein precursor to Rauscher leukemia virus gp69/71. Proc Natl Acad Sci U S A. 1976 Jul;73(7):2326–2330. doi: 10.1073/pnas.73.7.2326. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. O'Farrell P. H. High resolution two-dimensional electrophoresis of proteins. J Biol Chem. 1975 May 25;250(10):4007–4021. [PMC free article] [PubMed] [Google Scholar]
  22. O'Farrell P. Z., Goodman H. M. Resolution of simian virus 40 proteins in whole cell extracts by two-dimensional electrophoresis: heterogeneity of the major capsid protein. Cell. 1976 Oct;9(2):289–298. doi: 10.1016/0092-8674(76)90119-7. [DOI] [PubMed] [Google Scholar]
  23. Pal B. K., McAllister R. M., Gardner M. B., Roy-Burman P. Comparative studies on the structural phosphoproteins of mammalian type C viruses. J Virol. 1975 Jul;16(1):123–131. doi: 10.1128/jvi.16.1.123-131.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. 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]
  25. Strand M., August J. T. Structural proteins of ribonucleic acid tumor viruses. Purification of envelope, core, and internal components. J Biol Chem. 1976 Jan 25;251(2):559–564. [PubMed] [Google Scholar]
  26. 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]
  27. 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]
  28. 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]
  29. 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]

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

RESOURCES