Novel mutations that control the sphingolipid and cholesterol dependence of the semliki forest virus fusion protein

Prodyot K. Chatterjee, Christina H. Eng, Margaret Kielian

Research output: Contribution to journalArticle

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Abstract

The enveloped alphavirus Semliki Forest virus (SFV) infects cells via a membrane fusion reaction mediated by the E1 membrane protein. Efficient SFV-membrane fusion requires the presence of cholesterol and sphingolipid in the target membrane. Here we report on two mutants, srf-4 and srf-5, selected for growth in cholesterol-depleted cells. Like the previously isolated srf-3 mutant (E1 proline 226 to serine), the phenotypes of the srf-4 and srf-5 mutants were conferred by single-amino-acid changes in the E1 protein: leucine 44 to phenylalanine and valine 178 to alanine, respectively. Like srf-3, srf-4 and srf-5 show striking increases in the cholesterol independence of growth, infection, membrane fusion, and exit. Unexpectedly, and unlike srf-3, srf-4 and srf-5 showed highly efficient fusion with sphingolipid-free membranes in both lipid- and content-mixing assays. Both srf-4 and srf-5 formed E1 homotrimers of decreased stability compared to the homotrimers of the wild type and the srf-3 mutant. All three srf mutations lie in the same domain of E1, but the srf-4 and srf-5 mutations are spatially separated from srf-3. When expressed together, the three mutations could interact to produce increased sterol independence and to cause temperature-sensitive E1 transport. Thus, the srf-4 and srf-5 mutations identify novel regions of E1 that are distinct from the fusion peptide and srf-3 region and modulate the requirements for both sphingolipid and cholesterol in virus-membrane fusion.

Original languageEnglish (US)
Pages (from-to)12712-12722
Number of pages11
JournalJournal of Virology
Volume76
Issue number24
DOIs
StatePublished - Dec 2002

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viral fusion proteins
Semliki Forest virus
Viral Fusion Proteins
Semliki forest virus
sphingolipids
Sphingolipids
Cholesterol
cholesterol
mutation
Virus Internalization
mutants
Mutation
Membrane Fusion
Alphavirus
Membranes
Valine
Sterols
Growth
valine
Phenylalanine

ASJC Scopus subject areas

  • Immunology

Cite this

Novel mutations that control the sphingolipid and cholesterol dependence of the semliki forest virus fusion protein. / Chatterjee, Prodyot K.; Eng, Christina H.; Kielian, Margaret.

In: Journal of Virology, Vol. 76, No. 24, 12.2002, p. 12712-12722.

Research output: Contribution to journalArticle

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