Differential roles of two conserved glycine residues in the fusion peptide of Semliki Forest Virus

Swati Ghosh Shome, Margaret Kielian

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

Semliki Forest Virus (SFV) is an enveloped alphavirus that infects cells by a low-pH-dependent membrane fusion reaction. SFV fusion is catalyzed by the spike protein E1 subunit, which contains a putative fusion peptide between residues 79 and 97. Prior mutagenesis studies demonstrated that an E1 G91 D mutation blocks both virus-membrane fusion and the formation of a highly stable E1 trimer believed to be a critical fusion intermediate. We have here demonstrated that the G91D mutant was also inactive in hemifusion, suggesting that the E1 homotrimer is important in the initial stages of lipid mixing. Revertant analysis of a G91 deletion mutant indicated that G91 was crucial for the viability of SFV. In contrast, a G83D mutation produced infectious virus with both efficient fusion and homotrimer formation. Thus, the G83 position, although highly conserved among alphaviruses, was functional if replaced with a charged amino acid.

Original languageEnglish (US)
Pages (from-to)146-160
Number of pages15
JournalVirology
Volume279
Issue number1
DOIs
StatePublished - Jan 20 2001

Fingerprint

Semliki forest virus
Glycine
Alphavirus
Peptides
Virus Internalization
Mutation
Membrane Fusion
Protein Subunits
Mutagenesis
Viruses
Lipids
Amino Acids

Keywords

  • Alphavirus
  • Fusion peptide
  • In vitro transcription
  • Membrane fusion
  • Mutagenesis
  • Semliki Forest virus
  • Virus assembly
  • Virus entry
  • Virus fusion

ASJC Scopus subject areas

  • Virology
  • Infectious Diseases

Cite this

Differential roles of two conserved glycine residues in the fusion peptide of Semliki Forest Virus. / Shome, Swati Ghosh; Kielian, Margaret.

In: Virology, Vol. 279, No. 1, 20.01.2001, p. 146-160.

Research output: Contribution to journalArticle

@article{28e5feb10a88417086a26d873506afc7,
title = "Differential roles of two conserved glycine residues in the fusion peptide of Semliki Forest Virus",
abstract = "Semliki Forest Virus (SFV) is an enveloped alphavirus that infects cells by a low-pH-dependent membrane fusion reaction. SFV fusion is catalyzed by the spike protein E1 subunit, which contains a putative fusion peptide between residues 79 and 97. Prior mutagenesis studies demonstrated that an E1 G91 D mutation blocks both virus-membrane fusion and the formation of a highly stable E1 trimer believed to be a critical fusion intermediate. We have here demonstrated that the G91D mutant was also inactive in hemifusion, suggesting that the E1 homotrimer is important in the initial stages of lipid mixing. Revertant analysis of a G91 deletion mutant indicated that G91 was crucial for the viability of SFV. In contrast, a G83D mutation produced infectious virus with both efficient fusion and homotrimer formation. Thus, the G83 position, although highly conserved among alphaviruses, was functional if replaced with a charged amino acid.",
keywords = "Alphavirus, Fusion peptide, In vitro transcription, Membrane fusion, Mutagenesis, Semliki Forest virus, Virus assembly, Virus entry, Virus fusion",
author = "Shome, {Swati Ghosh} and Margaret Kielian",
year = "2001",
month = "1",
day = "20",
doi = "10.1006/viro.2000.0688",
language = "English (US)",
volume = "279",
pages = "146--160",
journal = "Virology",
issn = "0042-6822",
publisher = "Academic Press Inc.",
number = "1",

}

TY - JOUR

T1 - Differential roles of two conserved glycine residues in the fusion peptide of Semliki Forest Virus

AU - Shome, Swati Ghosh

AU - Kielian, Margaret

PY - 2001/1/20

Y1 - 2001/1/20

N2 - Semliki Forest Virus (SFV) is an enveloped alphavirus that infects cells by a low-pH-dependent membrane fusion reaction. SFV fusion is catalyzed by the spike protein E1 subunit, which contains a putative fusion peptide between residues 79 and 97. Prior mutagenesis studies demonstrated that an E1 G91 D mutation blocks both virus-membrane fusion and the formation of a highly stable E1 trimer believed to be a critical fusion intermediate. We have here demonstrated that the G91D mutant was also inactive in hemifusion, suggesting that the E1 homotrimer is important in the initial stages of lipid mixing. Revertant analysis of a G91 deletion mutant indicated that G91 was crucial for the viability of SFV. In contrast, a G83D mutation produced infectious virus with both efficient fusion and homotrimer formation. Thus, the G83 position, although highly conserved among alphaviruses, was functional if replaced with a charged amino acid.

AB - Semliki Forest Virus (SFV) is an enveloped alphavirus that infects cells by a low-pH-dependent membrane fusion reaction. SFV fusion is catalyzed by the spike protein E1 subunit, which contains a putative fusion peptide between residues 79 and 97. Prior mutagenesis studies demonstrated that an E1 G91 D mutation blocks both virus-membrane fusion and the formation of a highly stable E1 trimer believed to be a critical fusion intermediate. We have here demonstrated that the G91D mutant was also inactive in hemifusion, suggesting that the E1 homotrimer is important in the initial stages of lipid mixing. Revertant analysis of a G91 deletion mutant indicated that G91 was crucial for the viability of SFV. In contrast, a G83D mutation produced infectious virus with both efficient fusion and homotrimer formation. Thus, the G83 position, although highly conserved among alphaviruses, was functional if replaced with a charged amino acid.

KW - Alphavirus

KW - Fusion peptide

KW - In vitro transcription

KW - Membrane fusion

KW - Mutagenesis

KW - Semliki Forest virus

KW - Virus assembly

KW - Virus entry

KW - Virus fusion

UR - http://www.scopus.com/inward/record.url?scp=0035915997&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0035915997&partnerID=8YFLogxK

U2 - 10.1006/viro.2000.0688

DO - 10.1006/viro.2000.0688

M3 - Article

VL - 279

SP - 146

EP - 160

JO - Virology

JF - Virology

SN - 0042-6822

IS - 1

ER -