A system for functional analysis of Ebola virus glycoprotein

Ayato Takada, Clinton Robison, Hideo Goto, Anthony Sanchez, K. Gopal Murti, Michael Whitt, Yoshihiro Kawaoka

Research output: Contribution to journalArticle

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Abstract

Ebola virus causes hemorrhagic fever in humans and nonhuman primates, resulting in mortality rates of up to 90%. Studies of this virus have been hampered by its extraordinary pathogenicity, which requires biosafety level 4 containment. To circumvent this problem, we developed a novel complementation system for functional analysis of Ebola virus glycoproteins. It relies on a recombinant vesicular stomatitis virus (VSV) that contains the green fluorescent protein gene instead of the receptor-binding G protein gene (VSVΔG*). Herein we show that Ebola Reston virus glycoprotein (ResGP) is efficiently incorporated into VSV particles. This recombinant VSV with integrated ResGP (VSVΔG*-ResGP) infected primate cells more efficiently than any of the other mammalian or avian cells examined, in a manner consistent with the host range tropism of Ebola virus, whereas VSVΔG* complemented with VSV G protein (VSVΔG*-G) efficiently infected the majority of the cells tested. We also tested the utility of this system for investigating the cellular receptors for Ebola virus. Chemical modification of cells to alter their surface proteins markedly reduced their susceptibility to VSVΔG*-ResGP but not to VSVΔG*-G. These findings suggest that cell surface glycoproteins with N-linked oligosaccharide chains contribute to the entry of Ebola viruses, presumably acting as a specific receptor and/or cofactor for virus entry. Thus, our VSV system should be useful for investigating the functions of glycoproteins from highly pathogenic viruses or those incapable of being cultured in vitro.

Original languageEnglish (US)
Pages (from-to)14764-14769
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume94
Issue number26
DOIs
StatePublished - Dec 23 1997

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Ebolavirus
Glycoproteins
Vesicular Stomatitis
Viruses
Primates
Viral Tropism
Virus Internalization
Host Specificity
Membrane Glycoproteins
Green Fluorescent Proteins
Oligosaccharides
GTP-Binding Proteins
Virion
Genes
Virulence
Carrier Proteins
Membrane Proteins
Fever
Mortality

All Science Journal Classification (ASJC) codes

  • General

Cite this

A system for functional analysis of Ebola virus glycoprotein. / Takada, Ayato; Robison, Clinton; Goto, Hideo; Sanchez, Anthony; Murti, K. Gopal; Whitt, Michael; Kawaoka, Yoshihiro.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 94, No. 26, 23.12.1997, p. 14764-14769.

Research output: Contribution to journalArticle

Takada, Ayato ; Robison, Clinton ; Goto, Hideo ; Sanchez, Anthony ; Murti, K. Gopal ; Whitt, Michael ; Kawaoka, Yoshihiro. / A system for functional analysis of Ebola virus glycoprotein. In: Proceedings of the National Academy of Sciences of the United States of America. 1997 ; Vol. 94, No. 26. pp. 14764-14769.
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