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. 1980 Jul;35(1):41–51. doi: 10.1128/jvi.35.1.41-51.1980

Sequence relationship of glycosylated and unglycosylated gag polyproteins of Moloney murine leukemia virus.

S A Edwards, H Fan
PMCID: PMC288781  PMID: 6997511

Abstract

Both glycosylated and unglycosylated polyproteins coded by the gag gene are produced in cells infected with Moloney murine leukemia virus. GpP80gag is a glycosylated precursor of a larger gag glycoprotein exported to the cell surface, whereas Pr65gag is an unglycosylated precursor of the virion internal structural proteins. GpP80gag contains not only carbohydrate, but also additional polypeptide sequences not found in Pr65gag. In the experiment reported here, we localized the differences between GpP80gag and Pr65gag with respect to the domains of the individual gag proteins. This was done by comparison of partial proteolytic cleavage fragments from Pr65gag, from GpP80gag, and from the unglycosylated form of GpP80gag (P75gag) which had been immunoprecipitated by antisera specific for gag proteins p30, p15, and p10. We conclude that the additional polypeptide sequences in GpP80gag are located at or very near the amino terminus of the polyprotein. The carbohydrate in GpP80gag is attached to polypeptide sequences held in common between GpP80gag and Pr65gag.

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Selected References

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

  1. Berk A. J., Sharp P. A. Spliced early mRNAs of simian virus 40. Proc Natl Acad Sci U S A. 1978 Mar;75(3):1274–1278. doi: 10.1073/pnas.75.3.1274. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. 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]
  3. Celma M. L., Ehrenfeld E. Translation of poliovirus RNA in vitro: detection of two different initiation sites. J Mol Biol. 1975 Nov 15;98(4):761–780. doi: 10.1016/s0022-2836(75)80009-x. [DOI] [PubMed] [Google Scholar]
  4. Cleveland D. W., Fischer S. G., Kirschner M. W., Laemmli U. K. Peptide mapping by limited proteolysis in sodium dodecyl sulfate and analysis by gel electrophoresis. J Biol Chem. 1977 Feb 10;252(3):1102–1106. [PubMed] [Google Scholar]
  5. Crawford L. V., Cole C. N., Smith A. E., Paucha E., Tegtmeyer P., Rundell K., Berg P. Organization and expression of early genes of simian virus 40. Proc Natl Acad Sci U S A. 1978 Jan;75(1):117–121. doi: 10.1073/pnas.75.1.117. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Evans L. H., Dresler S., Kabat D. Synthesis and glycosylation of polyprotein precursors to the internal core proteins of Friend murine leukemia virus. J Virol. 1977 Dec;24(3):865–874. doi: 10.1128/jvi.24.3.865-874.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Fan H., Baltimore D. RNA metabolism of murine leukemia virus: detection of virus-specific RNA sequences in infected and uninfected cells and identification of virus-specific messenger RNA. J Mol Biol. 1973 Oct 15;80(1):93–117. doi: 10.1016/0022-2836(73)90235-0. [DOI] [PubMed] [Google Scholar]
  8. Gielkens A. L., Van Zaane D., Bloemers H. P., Bloemendal H. Synthesis of Rauscher murine leukemia virus-specific polypeptides in vitro. Proc Natl Acad Sci U S A. 1976 Feb;73(2):356–360. doi: 10.1073/pnas.73.2.356. [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. Kiely M. L., McKnight G. S., Schimke R. T. Studies on the attachment of carbohydrate to ovalbumin nascent chains in hen oviduct. J Biol Chem. 1976 Sep 25;251(18):5490–5495. [PubMed] [Google Scholar]
  11. 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]
  12. Kuo S. C., Lampen J. O. Tunicamycin--an inhibitor of yeast glycoprotein synthesis. Biochem Biophys Res Commun. 1974 May 7;58(1):287–295. doi: 10.1016/0006-291x(74)90925-5. [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. Ledbetter J. A. Two-dimensional analysis of murine leukemia virus gag-gene polyproteins. Virology. 1979 May;95(1):85–98. doi: 10.1016/0042-6822(79)90403-3. [DOI] [PubMed] [Google Scholar]
  16. Mains R. E., Eipper B. A., Ling N. Common precursor to corticotropins and endorphins. Proc Natl Acad Sci U S A. 1977 Jul;74(7):3014–3018. doi: 10.1073/pnas.74.7.3014. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Mueller-Lantzsch N., Fan H. Monospecific immunoprecipitation of murine leukemia virus polyribosomes: identification of p30 protein-specific messenger RNA. Cell. 1976 Dec;9(4 Pt 1):579–588. doi: 10.1016/0092-8674(76)90040-4. [DOI] [PubMed] [Google Scholar]
  18. 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]
  19. Pelham H. R., Jackson R. J. An efficient mRNA-dependent translation system from reticulocyte lysates. Eur J Biochem. 1976 Aug 1;67(1):247–256. doi: 10.1111/j.1432-1033.1976.tb10656.x. [DOI] [PubMed] [Google Scholar]
  20. Philipson L., Andersson P., Olshevsky U., Weinberg R., Baltimore D., Gesteland R. Translation of MuLV and MSV RNAs in nuclease-treated reticulocyte extracts: enhancement of the gag-pol polypeptide with yeast suppressor tRNA. Cell. 1978 Jan;13(1):189–199. doi: 10.1016/0092-8674(78)90149-6. [DOI] [PubMed] [Google Scholar]
  21. Roberts J. L., Herbert E. Characterization of a common precursor to corticotropin and beta-lipotropin: identification of beta-lipotropin peptides and their arrangement relative to corticotropin in the precursor synthesized in a cell-free system. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5300–5304. doi: 10.1073/pnas.74.12.5300. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Rothman J. E., Lodish H. F. Synchronised transmembrane insertion and glycosylation of a nascent membrane protein. Nature. 1977 Oct 27;269(5631):775–780. doi: 10.1038/269775a0. [DOI] [PubMed] [Google Scholar]
  23. Salomon R., Bar-Joseph M., Soreq H., Gozes I., Littauer U. Z. Translation in vitro of carnation mottle virus RNA. Regulatory function of the 3'-region. Virology. 1978 Oct 15;90(2):288–298. doi: 10.1016/0042-6822(78)90313-6. [DOI] [PubMed] [Google Scholar]
  24. Schultz A. M., Oroszlan S. Murine leukemia virus gag polyproteins: the peptide chain unique to Pr80 is located at the amino terminus. Virology. 1978 Dec;91(2):481–486. doi: 10.1016/0042-6822(78)90395-1. [DOI] [PubMed] [Google Scholar]
  25. Schultz A. M., Rabin E. H., Oroszlan S. Post-translational modification of Rauscher leukemia virus precursor polyproteins encoded by the gag gene. J Virol. 1979 Apr;30(1):255–266. doi: 10.1128/jvi.30.1.255-266.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Sefton B. M. Immediate glycosylation of Sindbis virus membrane proteins. Cell. 1977 Apr;10(4):659–668. doi: 10.1016/0092-8674(77)90099-x. [DOI] [PubMed] [Google Scholar]
  27. Stacey D. W., Allfrey V. G., Hanafusa H. Microinjection analysis of envelope-glycoprotein messenger activities of avian leukosis viral RNAs. Proc Natl Acad Sci U S A. 1977 Apr;74(4):1614–1618. doi: 10.1073/pnas.74.4.1614. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Westaway E. G. Strategy of the flavivirus genome: evidence for multiple internal initiation of translation of proteins specified by Kunjin virus in mammalian cells. Virology. 1977 Jul 15;80(2):320–335. doi: 10.1016/s0042-6822(77)80008-1. [DOI] [PubMed] [Google Scholar]
  29. 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]
  30. 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]
  31. Zaanie D., Gielkens A. L., Dekker-michielsen M. J., Bloemers H. P. Virus-specific precursor polypeptides in cells infected with Rauscher leukemia virus. Virology. 1975 Oct;67(2):544–552. doi: 10.1016/0042-6822(75)90454-7. [DOI] [PubMed] [Google Scholar]

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