1887

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Volume 96, Issue 7

Physico-chemical requirements and kinetics of membrane fusion of flavivirus-like particles Open Access

Abstract

Flaviviruses deliver their RNA genome into the host-cell cytoplasm by fusing their lipid envelope with a cellular membrane. Expression of the flavivirus pre-membrane and envelope glycoprotein genes in the absence of other viral genes results in the spontaneous assembly and secretion of virus-like particles (VLPs) with membrane fusion activity. Here, we examined the physico-chemical requirements for membrane fusion of VLPs from West Nile and Japanese encephalitis viruses. In a bulk fusion assay, optimal hemifusion (or lipid mixing) efficiencies were observed at 37 °C. Fusion efficiency increased with decreasing pH; half-maximal hemifusion was attained at pH 5.6. The anionic lipids bis(monoacylglycero)phosphate and phosphatidylinositol-3-phosphate, when present in the target membrane, significantly enhanced fusion efficiency, consistent with the emerging model that flaviviruses fuse with intermediate-to-late endosomal compartments, where these lipids are most abundant. In a single-particle fusion assay, VLPs catalysed membrane hemifusion, tracked as lipid mixing with the cellular membrane, on a timescale of 7–20 s after acidification. Lipid mixing kinetics suggest that hemifusion is a kinetically complex, multistep process.

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© 2015 The Authors
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2015-07-01
2024-04-24
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Physico-chemical requirements and kinetics of membrane fusion of flavivirus-like particles
J. Gen. Virol. 96, 1702 (2015); https://doi.org/10.1099/vir.0.000113
/content/journal/jgv/10.1099/vir.0.000113
/content/journal/jgv/10.1099/vir.0.000113
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