1887

Abstract

SUMMARY

We have previously demonstrated that Sindbis virus infection of Chinese hamster ovary (CHO) cells altered the protein glycosylation machinery of the cell, so that both normal, full-size (nine mannose-containing) oligosaccharides and abnormal, ‘truncated’ (five mannose-containing) oligosaccharides are transferred from lipid-linked precursors to newly synthesized viral membrane glycoproteins. In the present studies, we have examined the precursor oligosaccharides on viral glycoproteins that were pulse-labelled with [H]mannose in the presence or absence of glucose, since glucose starvation of uninfected CHO cells has been reported to induce synthesis of truncated precursor oligosaccharides. Pulse-labelling in the absence of glucose led to a greater than 10-fold increase in the relative amount of the truncated precursor oligosaccharides being transferred to the newly synthesized viral glycoproteins and to an apparent underglycosylation of some precursor viral polypeptides, with some asparaginyl sites not acquiring covalently linked oligosaccharides. The mature virion glycoproteins from CHO cells which were pulse-labelled in the absence of glucose and then ‘chased’ in the presence of glucose contained proportionately more unusual ManGlcNAc-size oligosaccharides. These small neutral-type oligosaccharides were apparently not as good a substrate for further processing into complex acidic-type oligosaccharides as the normal ManGlcNAc intermediate that results from the full-size precursor oligosaccharides.

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1985-07-01
2024-11-13
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References

  1. Bonatti S., Cancedda F. D. 1982; Posttranslational modifications of Sindbis virus glycoproteins: electrophoretic analysis of pulse-chase-labeled infected cells. Journal of Virology 42:64–70
    [Google Scholar]
  2. Bonner W. M., Laskey R. A. 1974; A film detection method for tritium-labeled proteins and nucleic acids in polyacrylamide gels. European Journal of Biochemistry 46:83–88
    [Google Scholar]
  3. Burke D., Keegstra K. 1979; Carbohydrate structure of Sindbis virus glycoprotein E2 from virus grown in hamster and chicken cells. Journal of Virology 29:546–554
    [Google Scholar]
  4. Chapman A., Li E., Kornfeld S. 1979a; The biosynthesis of the major lipid-linked oligosaccharide of Chinese hamster ovary cells occurs by the ordered addition of mannose residues. Journal of Biological Chemistry 254:10245–10249
    [Google Scholar]
  5. Chapman A., Trowbridge I. S., Hyman R., Kornfeld S. 1979b; Structure of the lipid-linked oligosaccharides that accumulate in class E Thy-1-negative mutant lymphomas. Cell 17:509–515
    [Google Scholar]
  6. Davidson S. K., Hunt L. A. 1983; Unusual neutral oligosaccharides in mature Sindbis virus glycoproteins are synthesized from truncated precursor oligosaccharides in Chinese hamster ovary cells. Journal of General Virology 64:613–625
    [Google Scholar]
  7. Duda E., Schlesinger M. J. 1975; Alterations in Sindbis viral envelope proteins by treating BHK cells with glucosamine. Journal of Virology 15:416–419
    [Google Scholar]
  8. Etchison L. R., Robertson J. S., Summers D. F. 1977; Partial structural analysis of the oligosaccharide moieties of the vesicular stomatitis virus glycoprotein by sequential chemical and enzymatic degradation. Virology 78:375–393
    [Google Scholar]
  9. Gershman H., Robbins P. W. 1981; Transitory effects of glucose starvation on the synthesis of dolichol-linked oligosaccharides in mammalian cells. Journal of Biological Chemistry 256:7774–7780
    [Google Scholar]
  10. Gibson R., Kornfeld S., Schlesinger S. 1981; The effect of oligosaccharide chains of different sizes on the maturation and physical properties of the G protein of vesicular stomatitis virus. Journal of Biological Chemistry 256:456–462
    [Google Scholar]
  11. Hakimi J., Atkinson P. H. 1982; Glycosylation of intracellular Sindbis virus glycoproteins. Biochemistry 21:2140–2145
    [Google Scholar]
  12. Hsieh P., Rosner M. R., Robbins P. W. 1983; Host-dependent variation of asparagine-linked oligosaccharides at individual glycosylation sites of Sindbis virus glycoproteins. Journal of Biological Chemistry 258:2548–2554
    [Google Scholar]
  13. Hubbard S. C., Ivatt R. J. 1981; Synthesis and processing of asparagine-linked oligosaccharides. Annual Review of Biochemistry 50:555–583
    [Google Scholar]
  14. Hunt L. A. 1981; Sindbis virus glycoproteins acquire unusual neutral oligosaccharides in both normal and lectin-resistant Chinese hamster ovary cell lines. Virology 113:534–543
    [Google Scholar]
  15. Hunt L. A., Summers D. F. 1976; Glycosylation of vesicular stomatitis virus glycoprotein in virus-infected HeLa cells. Journal of Virology 20:646–657
    [Google Scholar]
  16. Hunt L. A., Etchinson J. R., Summers D. F. 1978; Oligosaccharide chains are trimmed during synthesis of the envelope glycoprotein of vesicular stomatitis virus. Proceedings of the National Academy of Sciences, U.S.A 75:754–758
    [Google Scholar]
  17. Hunt L. A., Davidson S. K., Golemboski D. B. 1983; Unusual heterogeneity in the glycosylation of the G protein of the Hazelhurst strain of vesicular stomatitis virus. Archives of Biochemistry and Biophysics 226:347–356
    [Google Scholar]
  18. Kornfeld S., Gregory W., Chapman A. 1979; Class E Thy-1 negative mouse lymphoma cells utilize an alternate pathway of oligosaccharide processing to synthesize complex-type oligosaccharides. Journal of Biological Chemistry 254:11649–11654
    [Google Scholar]
  19. Laemmli U.K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, London 111:680–685
    [Google Scholar]
  20. Leavitt R., Schlesinger S., Kornfeld S. 1977a; Impaired intracellular migration and altered solubility of nonglycosylated glycoproteins of vesicular stomatitis virus and Sindbis virus. Journal of Biological Chemistry 252:9018–9023
    [Google Scholar]
  21. Leavitt R., Schlesinger S., Kornfeld S. 1977b; Tunicamycin inhibits glycosylation and multiplication of Sindbis and vesicular stomatitis viruses. Journal of Virology 21:375–385
    [Google Scholar]
  22. Narasimhan S., Stanley P., Schachter H. 1977; Control of glycoprotein synthesis. Lectin-resistant mutant containing only one of two distinct N-acetylglucosaminyltransferase activities present in wild type Chinese hamster ovary cells. Journal of Biological Chemistry 252:3926–3933
    [Google Scholar]
  23. Plummer T. H. Jr, Tarentino A. L. 1981; Facile cleavage of complex oligosaccharides by almond emulsion peptide: N-glycosidase. Journal of Biological Chemistry 256:10243–10246
    [Google Scholar]
  24. Rearick J. I., Chapman A., Kornfeld S. 1981; Glucose starvation alters lipid-linked oligosaccharide biosynthesis in Chinese hamster ovary cells. Journal of Biological Chemistry 256:6255–6261
    [Google Scholar]
  25. Rice C. M., Strauss J. H. 1981; Nucleotide sequence of the 26S mRN A of Sindbis virus and deduced sequence of the encoded virus structural proteins. Proceedings of the National Academy of Sciences, U.S.A 78:2062–2066
    [Google Scholar]
  26. Robertson J., Etchinson J., Summers D. F. 1976; Glycosylation sites of vesicular stomatitis virus glycoprotein. Journal of Virology 19:871–878
    [Google Scholar]
  27. Robertson M. A., Etchinson J. R., Robertson J. S., Summers D. F., Stanley P. 1978; Specific changes in the oligosaccharide moieties of VSV grown in different lectin-resistant CHO cells. Cell 13:515–526
    [Google Scholar]
  28. Schachter H., Narasimhan S., Gleeson P., Vella G. 1983; Glycosyltransferases involved in elongation of N- glycosidically linked oligosaccharides of the complex or N-acetyllactosamine type. Methods in Enzymology 98:98–134
    [Google Scholar]
  29. Schlesinger S., Schlesinger M. 1972; Formation of Sindbis virus proteins: identification of a precursor for one of the envelope proteins. Journal of Virology 10:925–932
    [Google Scholar]
  30. Schmidt M. F. G., Schlesinger M. J. 1980; Relation of fatty acid attachment to the translation and maturation of vesicular stomatitis and Sindbis virus membrane glycoproteins. Journal of Biological Chemistry 255:3334–3339
    [Google Scholar]
  31. Sefton B. M. 1977; Immediate glycosylation of Sindbis virus membrane proteins. Cell 10:659–668
    [Google Scholar]
  32. Stanley P. 1983; Selection of lectin-resistant mutants of animal cells. Methods in Enzymology 96:157–184
    [Google Scholar]
  33. Strauss J. H., Strauss E. G. 1977; Togaviruses. In The Molecular Biology of Animal Viruses vol 1: pp 111–166 Edited by Nayak D. P. New York: Marcel Dekker;
    [Google Scholar]
  34. Tabas I., Kornfeld S. 1979; Purification and characterization of a rat liver Golgi α-mannosidase capable of processing asparagine-linked oligosaccharides. Journal of Biological Chemistry 254:11655–11663
    [Google Scholar]
  35. Tarentino A. L., Plummer T. H. Jr, Maley F. 1974; The release of intact oligosaccharides from specific glycoproteins by endo-β-N-acetylglucosaminidase H. Journal of Biological Chemistry 249:818–824
    [Google Scholar]
  36. Trowbridge I. S., Hyman R. 1979; Abnormal lipid-linked oligosaccharides in class E Thy-1-negative mutant lymphomas. Cell 17:503–508
    [Google Scholar]
  37. Tulsiani D. R. P., Hubbard S. C., Robbins P. W., Touster O. 1982; α-d-mannosidases of rat Golgi membranes. Mannosidase II is the GlcNAc-Man5-cleaving enzyme in glycoprotein biosynthesis and mannosidases IA and IB are the enzymes converting Man9 precursors to Man5 intermediates. Journal of Biological Chemistry 257:3660–3668
    [Google Scholar]
  38. Welch W. J., Sefton B. M. 1979; Two small virus-specific polypeptides are produced during infection with Sindbis virus. Journal of Virology 29:1186–1195
    [Google Scholar]
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