Skip to main content
PMC home page
Journal of Virology logoLink to Journal of Virology
. 1991 Mar;65(3):1332–1339. doi: 10.1128/jvi.65.3.1332-1339.1991

Defective transport of Sindbis virus glycoproteins in End4 mutant Chinese hamster ovary cells.

J F Presley 1, R K Draper 1, D T Brown 1
PMCID: PMC239909  PMID: 1995947

Abstract

Mutant V.24.1, a temperature-sensitive derivative of Chinese hamster ovary cells, defines the End4 complementation group of mutants selected for resistance to protein toxins and has defective lysosomes at the restrictive temperature (P. A. Colbaugh, M. Stookey, and R. K. Draper, J. Cell Biol. 108:2211-2219, 1989). We have investigated the biosynthesis of Sindbis virus envelope glycoproteins in V.24.1 cells. When the cells were infected at the restrictive temperature, the envelope glycoproteins E1 and E2 were undetectable on the cell surface and proteolytic processing of the precursor protein pE2 to envelope protein E2 did not occur. Protein retained intracellularly was sensitive to endoglycosidase H and, by immunofluorescence localization, appeared to accumulate in the endoplasmic reticulum. We conclude that the genetic defect in V.24.1 cells impairs the transport of Sindbis virus glycoproteins, apparently at the level of export from the endoplasmic reticulum.

Full text

PDF
1332

Images in this article

1334Image
on p.1334
1334Image
on p.1334
1335Image
on p.1335
1336Image
on p.1336
1337Image
on p.1337

Selected References

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

  1. Balch W. E. Biochemistry of interorganelle transport. A new frontier in enzymology emerges from versatile in vitro model systems. J Biol Chem. 1989 Oct 15;264(29):16965–16968. [PubMed] [Google Scholar]
  2. 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]
  3. Colbaugh P. A., Kao C. Y., Shia S. P., Stookey M., Draper R. K. Three new complementation groups of temperature-sensitive Chinese hamster ovary cell mutants defective in the endocytic pathway. Somat Cell Mol Genet. 1988 Sep;14(5):499–507. doi: 10.1007/BF01534715. [DOI] [PubMed] [Google Scholar]
  4. Colbaugh P. A., Stookey M., Draper R. K. Impaired lysosomes in a temperature-sensitive mutant of Chinese hamster ovary cells. J Cell Biol. 1989 Jun;108(6):2211–2219. doi: 10.1083/jcb.108.6.2211. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Copeland C. S., Doms R. W., Bolzau E. M., Webster R. G., Helenius A. Assembly of influenza hemagglutinin trimers and its role in intracellular transport. J Cell Biol. 1986 Oct;103(4):1179–1191. doi: 10.1083/jcb.103.4.1179. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. EAGLE H. Amino acid metabolism in mammalian cell cultures. Science. 1959 Aug 21;130(3373):432–437. doi: 10.1126/science.130.3373.432. [DOI] [PubMed] [Google Scholar]
  7. Gething M. J., McCammon K., Sambrook J. Expression of wild-type and mutant forms of influenza hemagglutinin: the role of folding in intracellular transport. Cell. 1986 Sep 12;46(6):939–950. doi: 10.1016/0092-8674(86)90076-0. [DOI] [PubMed] [Google Scholar]
  8. Hashimoto K., Erdei S., Keränen S., Saraste J., Käriäinen L. Evidence for a separate signal sequence for the carboxy-terminal envelope glycoprotein E1 of Semliki forest virus. J Virol. 1981 Apr;38(1):34–40. doi: 10.1128/jvi.38.1.34-40.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hearing J., Gething M. J., Sambrook J. Addition of truncated oligosaccharides to influenza virus hemagglutinin results in its temperature-conditional cell-surface expression. J Cell Biol. 1989 Feb;108(2):355–365. doi: 10.1083/jcb.108.2.355. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hearing J., Hunter E., Rodgers L., Gething M. J., Sambrook J. Isolation of Chinese hamster ovary cell lines temperature conditional for the cell-surface expression of integral membrane glycoproteins. J Cell Biol. 1989 Feb;108(2):339–353. doi: 10.1083/jcb.108.2.339. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hsieh P., Rosner M. R., Robbins P. W. Selective cleavage by endo-beta-N-acetylglucosaminidase H at individual glycosylation sites of Sindbis virion envelope glycoproteins. J Biol Chem. 1983 Feb 25;258(4):2555–2561. [PubMed] [Google Scholar]
  12. Knipfer M. E., Brown D. T. Intracellular transport and processing of Sindbis virus glycoproteins. Virology. 1989 May;170(1):117–122. doi: 10.1016/0042-6822(89)90358-9. [DOI] [PubMed] [Google Scholar]
  13. Kreis T. E., Lodish H. F. Oligomerization is essential for transport of vesicular stomatitis viral glycoprotein to the cell surface. Cell. 1986 Sep 12;46(6):929–937. doi: 10.1016/0092-8674(86)90075-9. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Mann E., Edwards J., Brown D. T. Polycaryocyte formation mediated by Sindbis virus glycoproteins. J Virol. 1983 Mar;45(3):1083–1089. doi: 10.1128/jvi.45.3.1083-1089.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Nakano A., Nishijima M., Maeda M., Akamatsu Y. A temperature-sensitive Chinese hamster ovary cell mutant pleiotropically defective in protein export. Biochim Biophys Acta. 1985 Jun 30;845(3):324–332. doi: 10.1016/0167-4889(85)90195-8. [DOI] [PubMed] [Google Scholar]
  16. Novick P., Ferro S., Schekman R. Order of events in the yeast secretory pathway. Cell. 1981 Aug;25(2):461–469. doi: 10.1016/0092-8674(81)90064-7. [DOI] [PubMed] [Google Scholar]
  17. Novick P., Field C., Schekman R. Identification of 23 complementation groups required for post-translational events in the yeast secretory pathway. Cell. 1980 Aug;21(1):205–215. doi: 10.1016/0092-8674(80)90128-2. [DOI] [PubMed] [Google Scholar]
  18. Pfeffer S. R., Rothman J. E. Biosynthetic protein transport and sorting by the endoplasmic reticulum and Golgi. Annu Rev Biochem. 1987;56:829–852. doi: 10.1146/annurev.bi.56.070187.004145. [DOI] [PubMed] [Google Scholar]
  19. Presely J. F., Brown D. T. The proteolytic cleavage of PE2 to envelope glycoprotein E2 is not strictly required for the maturation of Sindbis virus. J Virol. 1989 May;63(5):1975–1980. doi: 10.1128/jvi.63.5.1975-1980.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Renz D., Brown D. T. Characteristics of Sindbis virus temperature-sensitive mutants in cultured BHK-21 and Aedes albopictus (Mosquito) cells. J Virol. 1976 Sep;19(3):775–781. doi: 10.1128/jvi.19.3.775-781.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Roff C. F., Fuchs R., Mellman I., Robbins A. R. Chinese hamster ovary cell mutants with temperature-sensitive defects in endocytosis. I. Loss of function on shifting to the nonpermissive temperature. J Cell Biol. 1986 Dec;103(6 Pt 1):2283–2297. doi: 10.1083/jcb.103.6.2283. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Scheefers H., Scheefers-Borchel U., Edwards J., Brown D. T. Distribution of virus structural proteins and protein-protein interactions in plasma membrane of baby hamster kidney cells infected with Sindbis or vesicular stomatitis virus. Proc Natl Acad Sci U S A. 1980 Dec;77(12):7277–7281. doi: 10.1073/pnas.77.12.7277. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Smith J. F., Brown D. T. Envelopments of Sindbis virus: synthesis and organization of proteins in cells infected with wild type and maturation-defective mutants. J Virol. 1977 Jun;22(3):662–678. doi: 10.1128/jvi.22.3.662-678.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Steinman R. M., Mellman I. S., Muller W. A., Cohn Z. A. Endocytosis and the recycling of plasma membrane. J Cell Biol. 1983 Jan;96(1):1–27. doi: 10.1083/jcb.96.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Wang R. H., Colbaugh P. A., Kao C. Y., Rutledge E. A., Draper R. K. Impaired secretion and fluid-phase endocytosis in the End4 mutant of Chinese hamster ovary cells. J Biol Chem. 1990 Nov 25;265(33):20179–20187. [PubMed] [Google Scholar]
  26. Wilson D. W., Wilcox C. A., Flynn G. C., Chen E., Kuang W. J., Henzel W. J., Block M. R., Ullrich A., Rothman J. E. A fusion protein required for vesicle-mediated transport in both mammalian cells and yeast. Nature. 1989 Jun 1;339(6223):355–359. doi: 10.1038/339355a0. [DOI] [PubMed] [Google Scholar]
  27. de Curtis I., Simons K. Dissection of Semliki Forest virus glycoprotein delivery from the trans-Golgi network to the cell surface in permeabilized BHK cells. Proc Natl Acad Sci U S A. 1988 Nov;85(21):8052–8056. doi: 10.1073/pnas.85.21.8052. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Virology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES