West Nile virus entry requires cholesterol-rich membrane microdomains and is independent of αvβ3 integrin

Guruprasad R. Medigeshi, Alec Hirsch, Daniel Streblow, Janko Nikolich-Zugich, Jay Nelson

Research output: Contribution to journalArticle

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Abstract

West Nile virus (WNV) has been the leading cause of viral encephalitis in the United States since 1999. The endocytic processes involved in the internalization of infectious WNV by various cell types are not well characterized, and the involvement of cholesterol-rich membrane microdomains, or lipid rafts, in the life cycle of WNV has not been investigated previously. In this study, we found that the depletion of cellular cholesterol levels by brief treatment with methyl-β-cyclodextrin resulted in a 100-fold reduction of the titers of infectious WNV released into the culture supernatant, as well as a reduction in the number of WNV genome copies in the cholesterol-depleted cells. The addition of exogenous cholesterol to cholesterol-depleted cells reversed this effect. Cholesterol depletion postinfection did not affect WNV growth, suggesting that the effect occurs at the level of WNV entry. We also showed that while WNV entry did not require αvβ3 integrin and focal adhesion kinase, WNV particles failed to be internalized by cholesterol-depleted cells. Finally, we showed the colocalization of the WNV envelope protein and cholera toxin B, which is internalized in a lipid raft-dependent pathway, in microdomain clusters at the plasma membrane. These data suggest that WNV utilizes lipid rafts during initial stages of internalization and that the lipid rafts may contain a factor(s) that may enhance WNV endocytosis.

Original languageEnglish (US)
Pages (from-to)5212-5219
Number of pages8
JournalJournal of Virology
Volume82
Issue number11
DOIs
StatePublished - Jun 2008

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Membrane Microdomains
West Nile virus
Virus Internalization
integrins
Integrins
Cholesterol
cholesterol
lipids
Lipids
non-specific protein-tyrosine kinase
viral encephalitis
cells
Viral Encephalitis
Viral Envelope Proteins
Focal Adhesion Protein-Tyrosine Kinases
cholera toxin
cyclodextrins
Cholera Toxin
Cyclodextrins
endocytosis

ASJC Scopus subject areas

  • Immunology

Cite this

West Nile virus entry requires cholesterol-rich membrane microdomains and is independent of αvβ3 integrin. / Medigeshi, Guruprasad R.; Hirsch, Alec; Streblow, Daniel; Nikolich-Zugich, Janko; Nelson, Jay.

In: Journal of Virology, Vol. 82, No. 11, 06.2008, p. 5212-5219.

Research output: Contribution to journalArticle

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