Effects of membrane potential and sphingolipid structures on fusion of Semliki Forest virus

Andrey V. Samsonov, Prodyot K. Chatterjee, Vladimir I. Razinkov, Christina H. Eng, Margaret Kielian, Fredric S. Cohen

Research output: Contribution to journalArticle

24 Citations (Scopus)

Abstract

Cells expressing the E1 and E2 envelope proteins of Semliki Forest virus (SFV) were fused to voltageclamped planar lipid bilayer membranes at low pH. Formation and evolution of fusion pores were electrically monitored by capacitance measurements, and membrane continuity was tracked by video fluorescence microscopy by including rhodamine-phosphatidylethanolamine in the bilayer. Fusion occurred without leakage for a negative potential applied to the trans side of the planar membrane. When a positive potential was applied, leakage was severe, obscuring the observation of any fusion. E1-mediated cell-cell fusion occurred without leakage for negative intracellular potentials but with substantial leakage for zero membrane potential. Thus, negative membrane potentials are generally required for nonleaky fusion. With planar bilayers as the target, the first fusion pore that formed almost always enlarged; pore flickering was a rare event. Similar to other target membranes, fusion required cholesterol and sphingolipids in the planar membrane. Sphingosine did not support fusion, but both ceramide, with even a minimal acyl chain (C2-ceramide), and lysosphingomyelin (lyso-SM) promoted fusion with the same kinetics. Thus, unrelated modifications to different parts of sphingosine yielded sphingolipids that supported fusion to the same degree. Fusion studies of pyrene-labeled SFV with cholesterol-containing liposomes showed that C2-ceramide supported fusion while lyso-SM did not, apparently due to its positive curvature effects. A model is proposed in which the hydroxyls of C-1 and C-3 as well as N of C-2 of the sphingosine backbone must orient so as to form multiple hydrogen bonds to amino acids of SFV E1 for fusion to proceed.

Original languageEnglish (US)
Pages (from-to)12691-12702
Number of pages12
JournalJournal of Virology
Volume76
Issue number24
DOIs
StatePublished - Dec 2002

Fingerprint

Semliki Forest virus
Semliki forest virus
sphingosine
sphingolipids
Sphingolipids
ceramides
membrane potential
Membrane Potentials
Sphingosine
Membranes
cholesterol
cell fusion
lipid bilayers
capacitance
Cholesterol
phosphatidylethanolamines
fluorescence microscopy
Video Microscopy
Membrane Fusion
hydrogen

ASJC Scopus subject areas

  • Immunology

Cite this

Effects of membrane potential and sphingolipid structures on fusion of Semliki Forest virus. / Samsonov, Andrey V.; Chatterjee, Prodyot K.; Razinkov, Vladimir I.; Eng, Christina H.; Kielian, Margaret; Cohen, Fredric S.

In: Journal of Virology, Vol. 76, No. 24, 12.2002, p. 12691-12702.

Research output: Contribution to journalArticle

Samsonov, Andrey V. ; Chatterjee, Prodyot K. ; Razinkov, Vladimir I. ; Eng, Christina H. ; Kielian, Margaret ; Cohen, Fredric S. / Effects of membrane potential and sphingolipid structures on fusion of Semliki Forest virus. In: Journal of Virology. 2002 ; Vol. 76, No. 24. pp. 12691-12702.
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