© The Rockefeller University Press,
0021-9525/1998//91 $5.00
The Journal of Cell Biology, Volume 140, Number 1,
, 1998 91-99
A Single Point Mutation Controls the Cholesterol Dependence of Semliki Forest Virus Entry and Exit
Malini Vashishtha,
Thomas Phalen,
Marianne T. Marquardt,
Jae S. Ryu,
Alice C. Ng, and
Margaret Kielian
Department of Cell Biology, Albert Einstein College of Medicine, Bronx, New York 10461
Membrane fusion and budding are key steps in the life cycle of all enveloped viruses. Semliki Forest virus (SFV) is an enveloped alphavirus that requires cellular membrane cholesterol for both membrane fusion and efficient exit of progeny virus from infected cells. We selected an SFV mutant, srf-3, that was strikingly independent of cholesterol for growth. This phenotype was conferred by a single amino acid change in the E1 spike protein subunit, proline 226 to serine, that increased the cholesterol independence of both srf-3 fusion and exit. The srf-3 mutant emphasizes the relationship between the role of cholesterol in membrane fusion and virus exit, and most significantly, identifies a novel spike protein region involved in the virus cholesterol requirement.
Abbreviations used in this paper: RT, reverse-transcriptase; SFV, Semliki Forest virus; wt, wild-type.
Address all correspondence to Margaret Kielian, Department of Cell Biology, Albert Einstein College of Medicine, 1300 Morris Park Avenue, Bronx, New York 10461. Tel.: (718) 430-3638. Fax: (718) 430-8574. E-mail: kielian{at}aecom.yu.edu
Marianne T. Marquardt's current address is Department of Biology, Stern College, New York, NY 10016.

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