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. 1979 Jan;29(1):285–292. doi: 10.1128/jvi.29.1.285-292.1979

Extracellular cleavage of the glycoprotein precursor of Rous sarcoma virus.

R Klemenz, H Diggelmann
PMCID: PMC353117  PMID: 219223

Abstract

The kinetics of cleavage of pr92gp, the precursor of the two glycoproteins of Rous sarcoma virus gp85 and gp35, were followed. Viral glycoproteins were detected by immunoprecipitation with anti-gp85 and anti-gp35 serum. It could be shown in pulse-chase experiments that little or no intracellular cleavage of the precursor took place during the time in which the majority of newly synthesized viral glycoprotein was exported from the cells. Soon after its synthesis, however, pr92gp underwent some modification that made it migrate slightly faster on sodium dodecyl sulfate-polyacrylamide gels. Under steady state conditions the precursor was shown to be the predominant form of intracellular viral glycoprotein. Virus which was harvested every 2 min from infected cells prelabeled for 90 min with [3H]mannose contained mostly uncleaved and only a little mature glycoprotein. By incubation of this freshly released virus in serum-free buffer, the majority of the glycoprotein precursor could be cleaved into mature gp85 and gp35. Virus harvested every 10 min contained only mature glycoproteins.

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Selected References

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

  1. BERNHARD W. Electron microscopy of tumor cells and tumor viruses; a review. Cancer Res. 1958 Jun;18(5):491–509. [PubMed] [Google Scholar]
  2. 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]
  3. Blobel G., Dobberstein B. Transfer of proteins across membranes. I. Presence of proteolytically processed and unprocessed nascent immunoglobulin light chains on membrane-bound ribosomes of murine myeloma. J Cell Biol. 1975 Dec;67(3):835–851. doi: 10.1083/jcb.67.3.835. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bolognesi D. P., Montelaro R. C., Frank H., Schäfer W. Assembly of type C oncornaviruses: a model. Science. 1978 Jan 13;199(4325):183–186. doi: 10.1126/science.202022. [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. Buchhagen D. L., Hanafusa H. Intracellular precursors to the major glycoprotein of avian oncoviruses in chicken embryo fibroblasts. J Virol. 1978 Mar;25(3):845–851. doi: 10.1128/jvi.25.3.845-851.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Canaani E., Helm K. V., Duesberg P. Evidence for 30-40S RNA as precursor of the 60-70S RNA of Rous sarcoma virus. Proc Natl Acad Sci U S A. 1973 Feb;70(2):401–405. doi: 10.1073/pnas.70.2.401. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Cheung K. S., Smith R. E., Stone M. P., Joklik W. K. Comparison of immature (rapid harvest) and mature Rous sarcoma virus particles. Virology. 1972 Dec;50(3):851–864. doi: 10.1016/0042-6822(72)90439-4. [DOI] [PubMed] [Google Scholar]
  9. Duesberg P., Canaani E., Von der Helm K. Evidence for 30-40S RNA as precursor of the 60-70S RNA of Rous sarcoma virus. Am J Clin Pathol. 1973 Jul;60(1):57–64. doi: 10.1093/ajcp/60.1.57. [DOI] [PubMed] [Google Scholar]
  10. Eisenman R., Vogt V. M., Diggelmann H. Synthesis of avian RNA tumor virus structural proteins. Cold Spring Harb Symp Quant Biol. 1975;39(Pt 2):1067–1075. doi: 10.1101/sqb.1974.039.01.122. [DOI] [PubMed] [Google Scholar]
  11. England J. M., Bolognesi D. P., Dietzschold B., Halpern M. S. Evidence that a precursor glycoprotein is cleaved to yield the major glycoprotein of avian tumor virus. J Virol. 1977 Feb;21(2):810–814. doi: 10.1128/jvi.21.2.810-814.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. 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]
  13. Klemenz R., Diggelmann H. The generation of the two envelope glycoproteins of Rous sarcoma virus from a common precursor polypeptide. Virology. 1978 Mar;85(1):63–74. doi: 10.1016/0042-6822(78)90411-7. [DOI] [PubMed] [Google Scholar]
  14. Korb J., Trávnícek M., Ríman J. The oncornavirus maturation process: quantitative correlation between morphological changes and conversion of genomic virion RNA. Intervirology. 1976;7(4-5):211–224. doi: 10.1159/000149954. [DOI] [PubMed] [Google Scholar]
  15. Leamnson R. N., Halpern M. S. Subunit structure of the glycoprotein complex of avian tumor virus. J Virol. 1976 Jun;18(3):956–968. doi: 10.1128/jvi.18.3.956-968.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Mirault M. E., Scherrer K. Isolation of preribosomes from HeLa cells and their characterization by electrophoresis on uniform and exponential-gradient-polyacrylamide gels. Eur J Biochem. 1971 Nov 11;23(2):372–386. doi: 10.1111/j.1432-1033.1971.tb01631.x. [DOI] [PubMed] [Google Scholar]
  17. Moelling K., Hayami M. Analysis of precursors to the envelope glycoproteins of avian RNA tumor viruses in chicken and quail cells. J Virol. 1977 Jun;22(3):598–607. doi: 10.1128/jvi.22.3.598-607.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Montelaro R. C., Sullivan S. J., Bolognesi D. P. An analysis of type-C retrovirus polypeptides and their associations in the virion. Virology. 1978 Jan;84(1):19–31. doi: 10.1016/0042-6822(78)90215-5. [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. Oppermann H., Bishop J. M., Varmus H. E., Levintow L. A joint produce of the genes gag and pol of avian sarcoma virus: a possible precursor of reverse transcriptase. Cell. 1977 Dec;12(4):993–1005. doi: 10.1016/0092-8674(77)90164-7. [DOI] [PubMed] [Google Scholar]
  21. Schlesinger M. J. Formation of an infectious virus-antibody complex with Rous sarcoma virus and antibodies directed against the major virus glycoprotein. J Virol. 1976 Mar;17(3):1063–1067. doi: 10.1128/jvi.17.3.1063-1067.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. 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]
  23. Shealy D. J., Rueckert R. R. Proteins of Rous-associated virus 61, an avian retrovirus: common precursor for glycoproteins gp85 and gp35 and use of pactamycin to map translational order of proteins in the gag, pol, and env genes. J Virol. 1978 May;26(2):380–388. doi: 10.1128/jvi.26.2.380-388.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Steinemann A., Stryer L. Accessibility of the carbohydrate moiety of rhodopsin. Biochemistry. 1973 Apr 10;12(8):1499–1502. doi: 10.1021/bi00732a005. [DOI] [PubMed] [Google Scholar]
  25. Titani K., Koide A., Hermann J., Ericsson L. H., Kumar S., Wade R. D., Walsh K. A., Neurath H., Fischer E. H. Complete amino acid sequence of rabbit muscle glycogen phosphorylase. Proc Natl Acad Sci U S A. 1977 Nov;74(11):4762–4766. doi: 10.1073/pnas.74.11.4762. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. 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]
  27. 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]
  28. Vogt V. M., Eisenman R. Identification of a large polypeptide precursor of avian oncornavirus proteins. Proc Natl Acad Sci U S A. 1973 Jun;70(6):1734–1738. doi: 10.1073/pnas.70.6.1734. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Witte O. N., Tsukamoto-Adey A., Weissman I. L. Cellular maturation of oncornavirus glycoproteins: topological arrangement of precursor and product forms in cellular membranes. Virology. 1977 Feb;76(2):539–553. doi: 10.1016/0042-6822(77)90236-7. [DOI] [PubMed] [Google Scholar]
  30. Witte O. N., Weissman I. L. Oncornavirus budding: kinetics of formation and utilization of viral membrane glycoprotein. Virology. 1976 Feb;69(2):464–473. doi: 10.1016/0042-6822(76)90477-3. [DOI] [PubMed] [Google Scholar]
  31. 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]
  32. 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]
  33. 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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