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. 1981 Sep 1;90(3):697–704. doi: 10.1083/jcb.90.3.697

Transient activity of Golgi-like membranes as donors of vesicular stomatitis viral glycoprotein in vitro

PMCID: PMC2111886  PMID: 6270159

Abstract

Previous reports demonstrated that the vesicular stomatitis viral glycoprotein (G protein), initially present in membranes of a Chinese hamster ovary mutant cell line (clone 15B) that is incapable of terminal glycosylation, can be transferred in vitro to exogenous Golgi membranes and there glycosylated (E. Fries and J. E. Rothman, 1980, Proc. Natl. Acad. Sci. U. S. A. 77:3870-3874; and J. E. Rothman and E. Fries, 1981, J. Cell Biol. 89:162-168). Here we present evidence that Golgi-like membranes serve as donors of G protein in this process. Pulse-chase experiments revealed that the donor activity of membranes is greatest at approximately 10 min of chase, a time when G protein has been shown to have arrived in Golgi stacks (J. E. Bergmann, K. T. Tokuyasu, and S. J. Singer, 1981, Proc. Natl. Acad. Sci. U. S. A. 78:1746-1750). Additional evidence that the G protein that is transferred to exogenous Golgi membranes in vitro had already entered the Golgi membranes in vivo was provided by observations that its oligosaccharides had already been trimmed, and that its distribution in a sucrose density gradient was coincident with that of enzymatic markers of Golgi membranes. The capacity of this Golgi-like membrane to serve as donor is transient, declining within 5 min after "trimming" in vivo as the G protein enters a "nontransferable" pool. The rapidity of the process suggests that both the "transferable" and "nontransferable" pools of G protein reside in Golgi-like membranes.

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Selected References

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  1. Bergeron J. J., Borts D., Cruz J. Passage of serum-destined proteins through the Golgi apparatus of rat liver. An examination of heavy and light Golgi fractions. J Cell Biol. 1978 Jan;76(1):87–97. doi: 10.1083/jcb.76.1.87. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bergmann J. E., Tokuyasu K. T., Singer S. J. Passage of an integral membrane protein, the vesicular stomatitis virus glycoprotein, through the Golgi apparatus en route to the plasma membrane. Proc Natl Acad Sci U S A. 1981 Mar;78(3):1746–1750. doi: 10.1073/pnas.78.3.1746. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bretz R., Bretz H., Palade G. E. Distribution of terminal glycosyltransferases in hepatic Golgi fractions. J Cell Biol. 1980 Jan;84(1):87–101. doi: 10.1083/jcb.84.1.87. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Brew K., Shaper J. H., Olsen K. W., Trayer I. P., Hill R. L. Cross-linking of the components of lactose synthetase with dimethylpimelimidate. J Biol Chem. 1975 Feb 25;250(4):1434–1444. [PubMed] [Google Scholar]
  5. Fleischer B., Fleischer S., Ozawa H. Isolation and characterization of Golgi membranes from bovine liver. J Cell Biol. 1969 Oct;43(1):59–79. doi: 10.1083/jcb.43.1.59. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Fries E., Rothman J. E. Transport of vesicular stomatitis virus glycoprotein in a cell-free extract. Proc Natl Acad Sci U S A. 1980 Jul;77(7):3870–3874. doi: 10.1073/pnas.77.7.3870. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Gottlieb C., Baenziger J., Kornfeld S. Deficient uridine diphosphate-N-acetylglucosamine:glycoprotein N-acetylglucosaminyltransferase activity in a clone of Chinese hamster ovary cells with altered surface glycoproteins. J Biol Chem. 1975 May 10;250(9):3303–3309. [PubMed] [Google Scholar]
  8. Hunt L. A., Etchison J. R., Summers D. F. Oligosaccharide chains are trimmed during synthesis of the envelope glycoprotein of vesicular stomatitis virus. Proc Natl Acad Sci U S A. 1978 Feb;75(2):754–758. doi: 10.1073/pnas.75.2.754. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hunt L. A., Summers D. F. Glycosylation of vesicular stomatitis virus glycoprotein in virus-infected HeLa cells. J Virol. 1976 Dec;20(3):646–657. doi: 10.1128/jvi.20.3.646-657.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Jelsema C. L., Morré D. J. Distribution of phospholipid biosynthetic enzymes among cell components of rat liver. J Biol Chem. 1978 Nov 10;253(21):7960–7971. [PubMed] [Google Scholar]
  11. Katz F. N., Lodish H. F. Transmembrane biogenesis of the vesicular stomatitis virus glycoprotein. J Cell Biol. 1979 Feb;80(2):416–426. doi: 10.1083/jcb.80.2.416. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Knipe D. M., Baltimore D., Lodish H. F. Separate pathways of maturation of the major structural proteins of vesicular stomatitis virus. J Virol. 1977 Mar;21(3):1128–1139. doi: 10.1128/jvi.21.3.1128-1139.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Knipe D. M., Lodish H. F., Baltimore D. Localization of two cellular forms of the vesicular stomatitis viral glycoprotein. J Virol. 1977 Mar;21(3):1121–1127. doi: 10.1128/jvi.21.3.1121-1127.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Kornfeld S., Li E., Tabas I. The synthesis of complex-type oligosaccharides. II. Characterization of the processing intermediates in the synthesis of the complex oligosaccharide units of the vesicular stomatitis virus G protein. J Biol Chem. 1978 Nov 10;253(21):7771–7778. [PubMed] [Google Scholar]
  15. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  16. Lenard J. Virus envelopes and plasma membranes. Annu Rev Biophys Bioeng. 1978;7:139–165. doi: 10.1146/annurev.bb.07.060178.001035. [DOI] [PubMed] [Google Scholar]
  17. Li E., Tabas I., Kornfeld S. The synthesis of complex-type oligosaccharides. I. Structure of the lipid-linked oligosaccharide precursor of the complex-type oligosaccharides of the vesicular stomatitis virus G protein. J Biol Chem. 1978 Nov 10;253(21):7762–7770. [PubMed] [Google Scholar]
  18. Reading C. L., Penhoet E. E., Ballou C. E. Carbohydrate structure of vesicular stomatitis virus glycoprotein. J Biol Chem. 1978 Aug 25;253(16):5600–5612. [PubMed] [Google Scholar]
  19. Robbins P. W., Hubbard S. C., Turco S. J., Wirth D. F. Proposal for a common oligosaccharide intermediate in the synthesis of membrane glycoproteins. Cell. 1977 Dec;12(4):893–900. doi: 10.1016/0092-8674(77)90153-2. [DOI] [PubMed] [Google Scholar]
  20. Rothman J. E., Bursztyn-Pettegrew H., Fine R. E. Transport of the membrane glycoprotein of vesicular stomatitis virus to the cell surface in two stages by clathrin-coated vesicles. J Cell Biol. 1980 Jul;86(1):162–171. doi: 10.1083/jcb.86.1.162. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Rothman J. E., Fries E. Transport of newly synthesized vesicular stomatitis viral glycoprotein to purified Golgi membranes. J Cell Biol. 1981 Apr;89(1):162–168. doi: 10.1083/jcb.89.1.162. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Schachter H., Jabbal I., Hudgin R. L., Pinteric L., McGuire E. J., Roseman S. Intracellular localization of liver sugar nucleotide glycoprotein glycosyltransferases in a Golgi-rich fraction. J Biol Chem. 1970 Mar 10;245(5):1090–1100. [PubMed] [Google Scholar]
  23. Tabas I., Kornfeld S. Purification and characterization of a rat liver Golgi alpha-mannosidase capable of processing asparagine-linked oligosaccharides. J Biol Chem. 1979 Nov 25;254(22):11655–11663. [PubMed] [Google Scholar]
  24. Tabas I., Kornfeld S. The synthesis of complex-type oligosaccharides. III. Identification of an alpha-D-mannosidase activity involved in a late stage of processing of complex-type oligosaccharides. J Biol Chem. 1978 Nov 10;253(21):7779–7786. [PubMed] [Google Scholar]
  25. Tabas I., Schlesinger S., Kornfeld S. Processing of high mannose oligosaccharides to form complex type oligosaccharides on the newly synthesized polypeptides of the vesicular stomatitis virus G protein and the IgG heavy chain. J Biol Chem. 1978 Feb 10;253(3):716–722. [PubMed] [Google Scholar]
  26. Tarentino A. L., Plummer T. H., Jr, Maley F. The release of intact oligosaccharides from specific glycoproteins by endo-beta-N-acetylglucosaminidase H. J Biol Chem. 1974 Feb 10;249(3):818–824. [PubMed] [Google Scholar]
  27. Tartakoff A. M. The Golgi complex: crossroads for vesicular traffic. Int Rev Exp Pathol. 1980;22:227–251. [PubMed] [Google Scholar]

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