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Journal of Virology, April 2002, p. 3267-3275, Vol. 76, No. 7
0022-538X/02/$04.00+0 DOI: 10.1128/JVI.76.7.3267-3275.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.
The Fusion Peptide of Semliki Forest Virus Associates with Sterol-Rich Membrane Domains
Anna Ahn, Don L. Gibbons, and Margaret Kielian*
Department of Cell Biology, Albert Einstein College of Medicine, Bronx, New York 10461
Received 28 August 2001/
Accepted 8 January 2001
Semliki Forest virus (SFV) is an enveloped alphavirus whose membrane fusion is triggered by low pH and promoted by cholesterol and sphingolipid in the target membrane. Fusion is mediated by E1, a viral membrane protein containing the putative fusion peptide. Virus mutant studies indicate that SFV's cholesterol dependence is controlled by regions of E1 outside of the fusion peptide. Both E1 and E1*, a soluble ectodomain form of E1, interact with membranes in a reaction dependent on low pH, cholesterol, and sphingolipid and form highly stable homotrimers. Here we have used detergent extraction and gradient floatation experiments to demonstrate that E1* associated selectively with detergent-resistant membrane domains (DRMs or rafts). In contrast, reconstituted full-length E1 protein or influenza virus fusion peptide was not associated with DRMs. Methyl ß-cyclodextrin quantitatively extracted both cholesterol and E1* from membranes in the absence of detergent, suggesting a strong association of E1* with sterol. Monoclonal antibody studies demonstrated that raft association was mediated by the proposed E1 fusion peptide. Thus, although other regions of E1 are implicated in the control of virus cholesterol dependence, once the SFV fusion peptide inserts in the target membrane it has a high affinity for membrane domains enriched in cholesterol and sphingolipid.
* Corresponding author. Mailing address: Dept. of Cell Biology, Albert Einstein College of Medicine, 1300 Morris Park Ave., Bronx, NY 10461. Phone: (718) 430-3638. Fax: (718) 430-8574. E-mail:
kielian{at}aecom.yu.edu.
Journal of Virology, April 2002, p. 3267-3275, Vol. 76, No. 7
0022-538X/02/$04.00+0 DOI: 10.1128/JVI.76.7.3267-3275.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.
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