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PMID: 11602759 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

PE2 cleavage mutants of Sindbis virus: correlation between viral infectivity and pH-dependent membrane fusion activation of the spike heterodimer.

Journal of virology ·Vol. 75 ·No. 22 ·2001-11-00 ·Pages 11196-204

Smit JM, Klimstra WB, Ryman KD, Bittman R, Johnston RE, Wilschut J

Abstract

The spike glycoprotein E2 of Sindbis virus (SIN) is synthesized in the infected cell as a PE2 precursor protein, which matures through cleavage by a cellular furin-like protease. Previous work has shown that SIN mutants impaired in PE2 cleavage are noninfectious on BHK-21 cells, the block in infection being localized at a step after virus-receptor interaction but prior to RNA replication. Here, we studied the membrane fusion properties of SIN PE2 cleavage mutants and observed that these viruses are impaired in their ability to form an E1 homotrimer and to fuse with liposomes at a mildly acidic pH. The block in spike rearrangement and fusion could be overridden by exposure of the mutant viruses to very low pH (<4.5). Cleavage mutants with second-site resuscitating mutations in PE2 were highly infectious for BHK-21 cells. The ability of these viruses to form E1 homotrimers and to fuse at a mildly acidic pH was completely restored despite a sustained lack of PE2 cleavage.

MeSH Terms
Animals Cell Line Cricetinae Dimerization Hydrogen-Ion Concentration Membrane Fusion Membrane Glycoproteins/chemistry Mutation Protein Precursors/chemistry Sindbis Virus/pathogenicity Viral Envelope Proteins/chemistry,metabolism
Chemicals
Membrane Glycoproteins Protein Precursors Viral Envelope Proteins glycoprotein E1, Sindbis virus glycoprotein E2, Sindbis virus
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Smit J M
Molecular Virology Section, Department of Medical Microbiology, University of Groningen, 9713 AV Groningen, The Netherlands.
Klimstra W B
Ryman K D
Bittman R
Johnston R E
Wilschut J
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Article Info
Journal
Journal of virology
Abbr.
J Virol
ISSN
0022-538X
DOI
10.1128/JVI.75.22.11196-11204.2001
Published
2001-11-00
Pages
11196-204
Language
English
Region
United States
NLM ID
0113724
PMCID
PMC114699
Subset
IM
Grants
NIAID NIH HHS · R01 AI022186 · United States
NHLBI NIH HHS · R01 HL016660 · United States
NHLBI NIH HHS · 2R01HL16660-27 · United States
NIAID NIH HHS · R01AI22186-14 · United States
External Links
PubMed source DOI: 10.1128/JVI.75.22.11196-11204.2001 PMC Full Text View journal details
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