Membrane fusion activity, oligomerization, and assembly of the rabies virus glycoprotein

Michael Whitt, Linda Buonocor, Christophe Prehaud, John K. Rose

Research output: Contribution to journalArticle

87 Citations (Scopus)

Abstract

The spike glycoprotein (G protein) of rabies virus (CVS strain) expressed in HeLa cells from cloned cDNA mediated membrane fusion after exposure to pHs of 6.1 or below. Chemical crosslinking showed that the rabies G protein, like the vesicular stomatitis virus (VSV) G protein, could be crosslinked to dimers and trimers, indicating that rabies G protein is a trimer. However, unlike the VSV G protein, rabies G protein trimers were not stable to sedimentation in sucrose gradients, even at a mildly acidic pH which stabilizes the VSV G protein trimers. In addition, we report that the expressed rabies virus G protein was functional because it could assemble into VSV particles (ts045) lacking VSV G protein and rescue infectivity. These VSV (rabies) pseudotypes were neutralized only by an antibody to the rabies G protein. We also examined the properties of a hybrid protein containing the extracellular domain of the rabies virus glycoprotein and the transmembrane and cytoplasmic domains of the VSV G protein. This protein was transported to tha cell surface and could be crosslinked to form dimers and trimers, but had little or no detectable membrane fusion activity. The lack of fusion activity was paradoxical because the hybrid protein could rescue VSV infectivity, although the titers were lower than those obtained with the wild-type rabies G protein.

Original languageEnglish (US)
Pages (from-to)681-688
Number of pages8
JournalVirology
Volume185
Issue number2
DOIs
StatePublished - Jan 1 1991
Externally publishedYes

Fingerprint

Rabies virus
Membrane Fusion
Glycoproteins
Vesicular Stomatitis
Rabies
Viruses
Proteins
HeLa Cells
Virion
Sucrose

All Science Journal Classification (ASJC) codes

  • Virology
  • Infectious Diseases

Cite this

Membrane fusion activity, oligomerization, and assembly of the rabies virus glycoprotein. / Whitt, Michael; Buonocor, Linda; Prehaud, Christophe; Rose, John K.

In: Virology, Vol. 185, No. 2, 01.01.1991, p. 681-688.

Research output: Contribution to journalArticle

Whitt, Michael ; Buonocor, Linda ; Prehaud, Christophe ; Rose, John K. / Membrane fusion activity, oligomerization, and assembly of the rabies virus glycoprotein. In: Virology. 1991 ; Vol. 185, No. 2. pp. 681-688.
@article{d857f820081a43d0b40ba409ef61f198,
title = "Membrane fusion activity, oligomerization, and assembly of the rabies virus glycoprotein",
abstract = "The spike glycoprotein (G protein) of rabies virus (CVS strain) expressed in HeLa cells from cloned cDNA mediated membrane fusion after exposure to pHs of 6.1 or below. Chemical crosslinking showed that the rabies G protein, like the vesicular stomatitis virus (VSV) G protein, could be crosslinked to dimers and trimers, indicating that rabies G protein is a trimer. However, unlike the VSV G protein, rabies G protein trimers were not stable to sedimentation in sucrose gradients, even at a mildly acidic pH which stabilizes the VSV G protein trimers. In addition, we report that the expressed rabies virus G protein was functional because it could assemble into VSV particles (ts045) lacking VSV G protein and rescue infectivity. These VSV (rabies) pseudotypes were neutralized only by an antibody to the rabies G protein. We also examined the properties of a hybrid protein containing the extracellular domain of the rabies virus glycoprotein and the transmembrane and cytoplasmic domains of the VSV G protein. This protein was transported to tha cell surface and could be crosslinked to form dimers and trimers, but had little or no detectable membrane fusion activity. The lack of fusion activity was paradoxical because the hybrid protein could rescue VSV infectivity, although the titers were lower than those obtained with the wild-type rabies G protein.",
author = "Michael Whitt and Linda Buonocor and Christophe Prehaud and Rose, {John K.}",
year = "1991",
month = "1",
day = "1",
doi = "10.1016/0042-6822(91)90539-N",
language = "English (US)",
volume = "185",
pages = "681--688",
journal = "Virology",
issn = "0042-6822",
publisher = "Academic Press Inc.",
number = "2",

}

TY - JOUR

T1 - Membrane fusion activity, oligomerization, and assembly of the rabies virus glycoprotein

AU - Whitt, Michael

AU - Buonocor, Linda

AU - Prehaud, Christophe

AU - Rose, John K.

PY - 1991/1/1

Y1 - 1991/1/1

N2 - The spike glycoprotein (G protein) of rabies virus (CVS strain) expressed in HeLa cells from cloned cDNA mediated membrane fusion after exposure to pHs of 6.1 or below. Chemical crosslinking showed that the rabies G protein, like the vesicular stomatitis virus (VSV) G protein, could be crosslinked to dimers and trimers, indicating that rabies G protein is a trimer. However, unlike the VSV G protein, rabies G protein trimers were not stable to sedimentation in sucrose gradients, even at a mildly acidic pH which stabilizes the VSV G protein trimers. In addition, we report that the expressed rabies virus G protein was functional because it could assemble into VSV particles (ts045) lacking VSV G protein and rescue infectivity. These VSV (rabies) pseudotypes were neutralized only by an antibody to the rabies G protein. We also examined the properties of a hybrid protein containing the extracellular domain of the rabies virus glycoprotein and the transmembrane and cytoplasmic domains of the VSV G protein. This protein was transported to tha cell surface and could be crosslinked to form dimers and trimers, but had little or no detectable membrane fusion activity. The lack of fusion activity was paradoxical because the hybrid protein could rescue VSV infectivity, although the titers were lower than those obtained with the wild-type rabies G protein.

AB - The spike glycoprotein (G protein) of rabies virus (CVS strain) expressed in HeLa cells from cloned cDNA mediated membrane fusion after exposure to pHs of 6.1 or below. Chemical crosslinking showed that the rabies G protein, like the vesicular stomatitis virus (VSV) G protein, could be crosslinked to dimers and trimers, indicating that rabies G protein is a trimer. However, unlike the VSV G protein, rabies G protein trimers were not stable to sedimentation in sucrose gradients, even at a mildly acidic pH which stabilizes the VSV G protein trimers. In addition, we report that the expressed rabies virus G protein was functional because it could assemble into VSV particles (ts045) lacking VSV G protein and rescue infectivity. These VSV (rabies) pseudotypes were neutralized only by an antibody to the rabies G protein. We also examined the properties of a hybrid protein containing the extracellular domain of the rabies virus glycoprotein and the transmembrane and cytoplasmic domains of the VSV G protein. This protein was transported to tha cell surface and could be crosslinked to form dimers and trimers, but had little or no detectable membrane fusion activity. The lack of fusion activity was paradoxical because the hybrid protein could rescue VSV infectivity, although the titers were lower than those obtained with the wild-type rabies G protein.

UR - http://www.scopus.com/inward/record.url?scp=0026339191&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0026339191&partnerID=8YFLogxK

U2 - 10.1016/0042-6822(91)90539-N

DO - 10.1016/0042-6822(91)90539-N

M3 - Article

VL - 185

SP - 681

EP - 688

JO - Virology

JF - Virology

SN - 0042-6822

IS - 2

ER -