A single-amino-acid substitution eliminates the stringent carbohydrate requirement for intracellular transport of a viral glycoprotein

A. M. Pitta, J. K. Rose, Carolyn E Machamer

Research output: Contribution to journalArticle

Abstract

In this report, we have investigated the contribution of primary sequence to the carbohydrate requirement for intracellular transport of two closely related glycoproteins, the G proteins of the San Juan and Orsay strains of vesicular stomatitis virus. We used site-directed mutagenesis of the coding sequence to eliminate the two consensus sites for glycosylation in the Orsay G protein. Whereas the nonglycosylated San Juan G protein required at least one of its two asparagine-linked oligosaccharides for transport to the plasma membrane at 37°C, a fraction of the Orsay G protein was transported without carbohydrate. Of the 10 amino acid differences between these two proteins, residue 172 (tyrosine in San Juan, aspartic acid in Orsay) played the major role in determining the stringency for the carbohydrate requirement. The rates at which the glycosylated and nonglycosylated Orsay G proteins were transported to the cell surface were the same, although a smaller fraction of the nonglycosylated protein was transported. These results suggest that the carbohydrate does not promote intracellular transport directly but influences a polypeptide folding or oligomerization step which is critical for transport.

Original languageEnglish (US)
Pages (from-to)3801-3809
Number of pages9
JournalJournal of Virology
Volume63
Issue number9
StatePublished - 1989
Externally publishedYes

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amino acid substitution
Amino Acid Substitution
G-proteins
GTP-Binding Proteins
glycoproteins
Glycoproteins
Carbohydrates
carbohydrates
Carbohydrate Sequence
Vesiculovirus
Vesicular Stomatitis
Asparagine
site-directed mutagenesis
asparagine
aspartic acid
glycosylation
Site-Directed Mutagenesis
Oligosaccharides
Glycosylation
Aspartic Acid

ASJC Scopus subject areas

  • Immunology

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A single-amino-acid substitution eliminates the stringent carbohydrate requirement for intracellular transport of a viral glycoprotein. / Pitta, A. M.; Rose, J. K.; Machamer, Carolyn E.

In: Journal of Virology, Vol. 63, No. 9, 1989, p. 3801-3809.

Research output: Contribution to journalArticle

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