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Characterization of the Functional Requirements of West Nile Virus Membrane Fusion

University Medical Center Groningen

¹ E-mail: jolanda.smit{at}med.umcg.nl

Flaviviruses infect their host cells by a membrane fusion reaction. In this study, we performed a functional analysis of the membrane fusion properties of West Nile virus (WNV) with liposomal target membranes. Membrane fusion was monitored continuously using a lipid mixing assay involving the fluorophore pyrene. Fusion of WNV with liposomes occurred on the time scale of seconds and was strictly dependent on mildly-acidic pH. Optimal fusion kinetics were observed at pH 6.3, the threshold for fusion being pH 6.9. Preincubation of the virus alone at pH 6.3 resulted in a rapid loss of fusion capacity. WNV fusion activity is strongly promoted by the presence of cholesterol in the target membrane. Furthermore, we provide direct evidence that cleavage of prM to M is a requirement for fusion activity of WNV.

Received 21 July 2009; accepted 11 October 2009.