A Single Point Mutation Controls the Cholesterol Dependence of Semliki Forest Virus Entry and Exit

doi:10.1083/jcb.140.1.91

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J. Cell Biol., Volume 140, Number 1, January 12, 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 envelopedalphavirus that requires cellular membrane cholesterol for bothmembrane fusion and efficient exit of progeny virus from infectedcells. We selected an SFV mutant, srf-3, that was strikingly independentof cholesterol for growth. This phenotype was conferred by a singleamino acid change in the E1 spike protein subunit, proline 226to serine, that increased the cholesterol independence of bothsrf-3 fusion and exit. The srf-3 mutant emphasizes the relationshipbetween the role of cholesterol in membrane fusion and virus exit,and most significantly, identifies a novel spike protein regioninvolved in the virus cholesterol requirement.

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