Expression from cloned cDNA of cell-surface secreted forms of the glycoprotein of vesicular stomatitis virus in eucaryotic cells

Rose, John K.; Bergmann, John E.

doi:10.1016/0092-8674(82)90280-x

Cited by 263 publications

(175 citation statements)

References 27 publications

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“…6A) and the pattern of internal labeling in the same cell (Fig. 6B) (5,6). Also, the pattern of internal labeling, showing intense labeling of the perinuclear Golgi region, is typical of normal G protein (6 …”

Section: Methodsmentioning

confidence: 79%

“…The mutated gene was then expressed in COS cells (5). This single amino acid substitution eliminated addition of palmitate to the G protein.…”

Section: Discussionmentioning

confidence: 99%

“…By expressing a cDNA clone encoding the G protein from a simian virus 40 vector in eukaryotic cells (5), we were able to show that the protein was synthesized, glycosylated, and transported to the plasma membrane just as in VSV-infected cells. In a subsequent study, we analyzed expression of truncated genes encoding G proteins lacking portions of the carboxyl-terminal cytoplasmic domain.…”

mentioning

confidence: 99%

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The presence of cysteine in the cytoplasmic domain of the vesicular stomatitis virus glycoprotein is required for palmitate addition.

Rose¹,

Adams²,

Gallione³

1984

Proc. Natl. Acad. Sci. U.S.A.

158

109

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Section: Methodsmentioning

confidence: 79%

“…The mutated gene was then expressed in COS cells (5). This single amino acid substitution eliminated addition of palmitate to the G protein.…”

Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

See 1 more Smart Citation

The presence of cysteine in the cytoplasmic domain of the vesicular stomatitis virus glycoprotein is required for palmitate addition.

Rose¹,

Adams²,

Gallione³

1984

Proc. Natl. Acad. Sci. U.S.A.

158

109

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“…To determine the rate and efficiency of trimerization, we expressed wt and mutant G proteins in COS cells by transfection with the appropriate plasmid DNA (Rose and Bergmann, 1982). The cells were pulsed with [35S]methionine and chased in the presence of excess cold methionine for various times.…”

Section: Resultsmentioning

confidence: 99%

Differential effects of mutations in three domains on folding, quaternary structure, and intracellular transport of vesicular stomatitis virus G protein.

Doms

Ruusala

Machamer

et al. 1988

The Journal of Cell Biology

193

148

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Abstract. The vesicular stomatitis virus glycoprotein (G protein) is an integral membrane protein which assembles into noncovalently associated trimers before transport from the endoplasmic reticulum. In this study we have examined the folding and oligomeric assembly of twelve mutant G proteins with alterations in the cytoplasmic, transmembrane, or ectodomains. Through the use of conformation-specific antibodies, we found that newly synthesized G protein folded into a conformation similar to the mature form within 1-3 min of synthesis and before trimer formation. Mutant proteins not capable of undergoing correct initial folding did not trimerize, were not transported, and were found in large aggregates. They had, as a rule, mutations in the ectodomain, including several with altered glycosylation patterns. In contrast, mutations in the cytoplasmic domain generally had little effect on folding and trimerization. These mutant proteins, whose ectodomains were identical to the wild-type by several assays, were either transported to the cell surface slowly or not at all. We concluded that while correct ectodomain folding and trimer formation are prerequisites for transport, they alone are not sufficient. The results suggest that the cytoplasmic domain of the wild-type protein may facilitate rapid, efficient transport from the ER, which can be easily affected or eliminated by tail mutations that do not detectably affect the ectodomain.

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“…The expression plasmid pSV2G (35) was provided by Dr. John Rose, Salk Institute. The vector did not yield useful levels of expression in MDCK cells, presumably due to the presence of the small t intron (10,15).…”

Section: Construction Of Expression Plasmidsmentioning

confidence: 99%

Sorting and endocytosis of viral glycoproteins in transfected polarized epithelial cells.

Gottlieb¹,

González²,

Rizzolo³

et al. 1986

The Journal of Cell Biology

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Expression from cloned cDNA of cell-surface secreted forms of the glycoprotein of vesicular stomatitis virus in eucaryotic cells

Cited by 263 publications

References 27 publications

The presence of cysteine in the cytoplasmic domain of the vesicular stomatitis virus glycoprotein is required for palmitate addition.

The presence of cysteine in the cytoplasmic domain of the vesicular stomatitis virus glycoprotein is required for palmitate addition.

Differential effects of mutations in three domains on folding, quaternary structure, and intracellular transport of vesicular stomatitis virus G protein.

Sorting and endocytosis of viral glycoproteins in transfected polarized epithelial cells.

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