Glycoprotein-dependent acidification of vesicular stomatitis virus enhances release of matrix protein

Chad Mire, Derek Dube, Sue E. Delos, Judith M. White, Michael A. Whitt

Research output: Contribution to journalArticle

26 Citations (Scopus)

Abstract

To study vesicular stomatitis virus (VSV) entry and uncoating, we generated a recombinant VSV encoding a matrix (M) protein containing a C-terminal tetracysteine Lumio tag (rVSV-ML) that could be fluorescently labeled using biarsenical compounds. Quantitative confocal microscopy showed that there is a transient loss of fluorescence at early times after the initiation of endocytosis of rVSV-ML-Green (rVSV-MLG) virions, which did not occur when cells were treated with bafilomycin A1. The reduction in fluorescence occurred 5 to 10 min postentry, followed by a steady increase in fluorescence intensity from 15 to 60 min postentry. A similar loss of fluorescence was observed in vitro when virions were exposed to acidic pH. The reduction in fluorescence required G protein since "bald" ΔG-MLG particles did not show a similar loss of fluorescence at low pH. Based on the pH-dependent fluorescence properties of Lumio Green, we hypothesize that the loss of fluorescence of rVSV-MLG virions during virus entry is due to a G ectodomain-dependent acidification of the virion interior. Biochemical analysis indicated that low pH also resulted in an enhancement of M protein dissociation from partially permeabilized, but otherwise intact, wild-type virions. From these data we propose that low-pH conformational changes in G protein promote acidification of the virus interior, which facilitates the release of M from ribonucleoprotein particles during uncoating.

Original languageEnglish (US)
Pages (from-to)12139-12150
Number of pages12
JournalJournal of Virology
Volume83
Issue number23
DOIs
StatePublished - Dec 2009
Externally publishedYes

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Virus Release
Vesiculovirus
Vesicular Stomatitis
acidification
glycoproteins
Glycoproteins
Fluorescence
fluorescence
virion
Virion
Proteins
proteins
Virus Internalization
G-proteins
GTP-Binding Proteins
Virus Uncoating
Viruses
ribonucleoproteins
viruses
Ribonucleoproteins

ASJC Scopus subject areas

  • Immunology
  • Virology

Cite this

Glycoprotein-dependent acidification of vesicular stomatitis virus enhances release of matrix protein. / Mire, Chad; Dube, Derek; Delos, Sue E.; White, Judith M.; Whitt, Michael A.

In: Journal of Virology, Vol. 83, No. 23, 12.2009, p. 12139-12150.

Research output: Contribution to journalArticle

Mire, Chad ; Dube, Derek ; Delos, Sue E. ; White, Judith M. ; Whitt, Michael A. / Glycoprotein-dependent acidification of vesicular stomatitis virus enhances release of matrix protein. In: Journal of Virology. 2009 ; Vol. 83, No. 23. pp. 12139-12150.
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