Glycoprotein cytoplasmic domain sequences required for rescue of a vesicular stomatitis virus glycoprotein mutant

Michael Whitt, L. Chong, J. K. Rose

Research output: Contribution to journalArticle

82 Citations (Scopus)

Abstract

We have used transient expression of the wild-type vesicular stomatitis virus (VSV) glycoprotein (G protein) from cloned cDNA to rescue a temperature-sensitive G protein mutant of VSV in cells at the nonpermissive temperature. Using cDNAs encoding G proteins with deletions in the normal 29-amino-acid cytoplasmic domain, we determined that the presence of either the membrane-proximal 9 amino acids or the membrane-distal 12 amino acids was sufficient for rescue of the temperature-sensitive mutant. G proteins with cytoplasmic domains derived from other cellular or viral G proteins did not rescue the mutant, nor did G proteins with one or three amino acids of the normal cytoplasmic domain. Rescue correlated directly with the ability of the G proteins to be incorporated into virus particles. This was shown by analysis of radiolabeled particles separated on sucrose gradients as well as by electron microscopy of rescued virus after immunogold labeling. Quantitation of surface expression showed that all of the mutated G proteins were expressed less efficiently on the cell surface than was wild-type G protein. However, we were able to correct for differences in rescue efficiency resulting from differences in the level of surface expression by reducing wild-type G protein expression to levels equivalent to those observed for the mutated G proteins. Our results provide evidence that at least a portion of the cytoplasmic domain is required for efficient assembly of the VSV G protein into virions during virus budding.

Original languageEnglish (US)
Pages (from-to)3569-3578
Number of pages10
JournalJournal of Virology
Volume63
Issue number9
StatePublished - 1989
Externally publishedYes

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Vesiculovirus
Vesicular Stomatitis
G-proteins
GTP-Binding Proteins
glycoproteins
Glycoproteins
Viruses
mutants
Amino Acids
amino acids
virion
Virion
Temperature
Complementary DNA
Virus Release
viruses
temperature
Membranes
Viral Proteins
Sucrose

All Science Journal Classification (ASJC) codes

  • Immunology

Cite this

Glycoprotein cytoplasmic domain sequences required for rescue of a vesicular stomatitis virus glycoprotein mutant. / Whitt, Michael; Chong, L.; Rose, J. K.

In: Journal of Virology, Vol. 63, No. 9, 1989, p. 3569-3578.

Research output: Contribution to journalArticle

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