Negative potentials across biological membranes promote fusion by class II and class III viral proteins

Mol Biol Cell. 2010 Jun 15;21(12):2001-12. doi: 10.1091/mbc.e09-10-0904. Epub 2010 Apr 28.

Abstract

Voltage was investigated as a factor in the fusion of virions. Virions, pseudotyped with a class II, SFV E1 or VEEV E, or a class III protein, VSV G, were prepared with GFP within the core and a fluorescent lipid. This allowed both hemifusion and fusion to be monitored. Voltage clamping the target cell showed that fusion is promoted by a negative potential and hindered by a positive potential. Hemifusion occurred independent of polarity. Lipid dye movement, in the absence of content mixing, ceased before complete transfer for positive potentials, indicating that reversion of hemifused membranes into two distinct membranes is responsible for voltage dependence and inhibition of fusion. Content mixing quickly followed lipid dye transfer for a negative potential, providing a direct demonstration that hemifusion induced by class II and class III viral proteins is a functional intermediate of fusion. In the hemifused state, virions that fused exhibited slower lipid transfer than did nonfusing virions. All viruses with class II or III fusion proteins may utilize voltage to achieve infection.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Cell Fusion
  • Cell Line
  • Cell Membrane / physiology*
  • Electricity
  • Fluorescence
  • Fluorescent Dyes / metabolism
  • Humans
  • Membrane Fusion / physiology*
  • Membrane Potentials / physiology*
  • Time Factors
  • Viral Fusion Proteins / metabolism*
  • Virion / metabolism

Substances

  • Fluorescent Dyes
  • Viral Fusion Proteins