Alphavirus 6K proteins form ion channels

J Biol Chem. 2002 Dec 6;277(49):46923-31. doi: 10.1074/jbc.M207847200. Epub 2002 Sep 12.

Abstract

Ross River virus and Barmah Forest virus are Australian arboviruses of the Alphavirus genus. Features of alphavirus infection include an increased permeability of cells to monovalent cations followed by virion budding. Virally encoded ion channels are thought to have a role in these processes. In this paper, the 6K proteins of Ross River virus and Barmah Forest virus are shown to form cation-selective ion channels in planar lipid bilayers. Using a novel purification method, bacterially expressed 6K proteins were inserted into bilayers with a defined orientation (i.e. N-terminal cis, C-terminal trans). Channel activity was reversibly inhibited by antibodies to the N and C termini of 6K protein added to the cis and trans baths, respectively. Channel conductances varied from 40-800 picosiemens, suggesting that the protein is able to form channels with a range of possible oligomerization states.

MeSH terms

  • Alphavirus / metabolism*
  • Amino Acid Sequence
  • Blotting, Western
  • Cloning, Molecular
  • DNA, Complementary / metabolism
  • Electrophoresis, Polyacrylamide Gel
  • Electrophysiology
  • Escherichia coli / metabolism
  • Glutathione Transferase / metabolism
  • Ion Channels / chemistry*
  • Ions / metabolism
  • Lipid Bilayers / metabolism
  • Molecular Sequence Data
  • Mutagenesis, Site-Directed
  • Mutation
  • Peptides / chemistry
  • Point Mutation
  • Protein Structure, Tertiary
  • Recombinant Fusion Proteins / metabolism
  • Sequence Homology, Amino Acid
  • Time Factors
  • Viral Proteins / chemistry

Substances

  • DNA, Complementary
  • Ion Channels
  • Ions
  • Lipid Bilayers
  • Peptides
  • Recombinant Fusion Proteins
  • Viral Proteins
  • Glutathione Transferase