Skip to main content
PMC home page
Journal of Virology logoLink to Journal of Virology
. 1979 Jun;30(3):799–804. doi: 10.1128/jvi.30.3.799-804.1979

Biosynthesis of an unglycosylated envelope glycoprotein of Rous sarcoma virus in the presence of tunicamycin.

H Diggelmann
PMCID: PMC353390  PMID: 90167

Abstract

Cells stably infected with Rous sarcoma virus were treated with tunicamycin to prevent the glycosylation of the precursor (pr92gp) to the two viral envelope glycoproteins gp85 and gp35. Pretreatment of the cells for 4 h with the antibiotic resulted in a 90% reduction in [3H]mannose incorporation into total cellular glycoproteins, intracellular viral glycoproteins, and released virus particles. Protein synthesis and virus particle formation were not significantly affected by the treatment. A new polypeptide made in the presence of the drug was identified by immunoprecipitation of pulse-labeled cell lysates with monospecific anti-gp85 and anti-gp35 sera. This polypeptide, migrating on sodium dodecyl sulfate-polyacrylamide gels as a molecule of 62,000 daltons (pr62), contained no [3H]mannose, was labeled with [S35]methionine and [3H]arginine, could not be chased into the higher-molecular-weight glycosylated form, and contained the same [3H]arginine tryptic peptides as pr92gp. The unglycosylated pr62 was still detectable 2 h after the pulse labeling of the cells. The lack of glycosylation of pr62 did not seem to reduce its stability. No clear evidence for the incorporation of this molecule or its cleavage products into viral particles could be obtained. To code for an envelope polypeptide of 62,000 daltons, only about 1,500 nucleotides or 15% of the total coding capacity of the virus are needed.

Full text

PDF
799

Images in this article

801Image
on p.801

Selected References

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

  1. 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]
  2. Eisenman R., Shaikh R., Mason W. S. Identification of an avian oncovirus polyprotein in uninfected chick cells. Cell. 1978 May;14(1):89–104. doi: 10.1016/0092-8674(78)90304-5. [DOI] [PubMed] [Google Scholar]
  3. 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]
  4. Garoff H., Schwarz R. T. Glycosylation is not necessary for membrane insertion and cleavage of Semliki Forest virus membrane proteins. Nature. 1978 Aug 3;274(5670):487–490. doi: 10.1038/274487a0. [DOI] [PubMed] [Google Scholar]
  5. Hemming F. W. Dolichol phosphate, a coenzyme in the glycosylation of animal membrane-bound glycoproteins. Biochem Soc Trans. 1977;5(4):1223–1231. doi: 10.1042/bst0051223. [DOI] [PubMed] [Google Scholar]
  6. Klemenz R., Diggelmann H. Extracellular cleavage of the glycoprotein precursor of Rous sarcoma virus. J Virol. 1979 Jan;29(1):285–292. doi: 10.1128/jvi.29.1.285-292.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. 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]
  8. 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]
  9. Krantz M. J., Lee Y. C., Hung P. P. Characterization and comparison of the major glycoprotein from three strains of Rous sarcoma virus. Arch Biochem Biophys. 1976 May;174(1):66–73. doi: 10.1016/0003-9861(76)90324-6. [DOI] [PubMed] [Google Scholar]
  10. Lewandowski L. J., Smith R. E., Bolognesi D. P., Halpern M. S. Viral glycoprotein synthesis under conditions of glucosamine block in cells transformed by avian sarcoma viruses. Virology. 1975 Aug;66(2):347–355. doi: 10.1016/0042-6822(75)90208-1. [DOI] [PubMed] [Google Scholar]
  11. 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]
  12. Schwarz R. T., Rohrschneider J. M., Schmidt M. F. Suppression of glycoprotein formation of Semliki Forest, influenza, and avian sarcoma virus by tunicamycin. J Virol. 1976 Sep;19(3):782–791. doi: 10.1128/jvi.19.3.782-791.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. 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]
  14. Struck D. K., Lennarz W. J. Evidence for the participation of saccharide-lipids in the synthesis of the oligosaccharide chain of ovalbumin. J Biol Chem. 1977 Feb 10;252(3):1007–1013. [PubMed] [Google Scholar]
  15. Tkacz J. S., Lampen O. Tunicamycin inhibition of polyisoprenyl N-acetylglucosaminyl pyrophosphate formation in calf-liver microsomes. Biochem Biophys Res Commun. 1975 Jul 8;65(1):248–257. doi: 10.1016/s0006-291x(75)80086-6. [DOI] [PubMed] [Google Scholar]
  16. 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