A glycine to alanine substitution in the paramyxovirus SV5 fusion peptide increases the initial rate of fusion

Sangeeta Bagai, Robert A. Lamb*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

11 Scopus citations

Abstract

Simian virus 5 fusion (F) protein mutant F-G3A, which contains a glycine-to-alanine substitution at position 3 in the conserved hydrophobic fusion peptide at the N-terminus of the F1 subunit, has been shown previously to cause increased syncytium formation compared to wild-type (wt) F protein, when expressed using an SV40 recombinant virus vector system (C. M. Horvath and R. A. Lamb (1992) J. Virol. 66, 2443-2455). The wt F and the F-G3A proteins were expressed in eukaryotic cells using the vaccinia virus- bacteriophage T7 RNA polymerase (vac-T7) expression system, and they showed similar cell surface expression levels as determined by flow cytometry. The final extent of fusion when the vac-T7 expression system was used was not found to be greatly different when examined with a reporter gene activation assay. However, the initial rate of fusion was found to be five- to sixfold higher for the F-G3A mutant protein than the wt F protein, when examined using a quantitative assay for lipid mixing based on relief of self-quenching of fluorescence of the lipid probe octadecyl rhodamine (R18). A microscopic fluorescent dye transfer assay also showed a much earlier spread of dye from R18-labeled red blood cells to the cells expressing the mutant F-G3A protein than the wt F protein. Thus, these data indicate that a single gly-to-ala mutation in the fusion peptide domain, although not affecting the final extent of fusion, significantly increased the rate of fusion. Possible mechanisms for the increased rate of fusion are discussed.

Original languageEnglish (US)
Pages (from-to)283-290
Number of pages8
JournalVirology
Volume238
Issue number2
DOIs
StatePublished - Nov 24 1997

ASJC Scopus subject areas

  • Virology

Fingerprint Dive into the research topics of 'A glycine to alanine substitution in the paramyxovirus SV5 fusion peptide increases the initial rate of fusion'. Together they form a unique fingerprint.

Cite this