The cytoplasmic domain of influenza M2 protein interacts with caveolin-1

Arch Biochem Biophys. 2009 Jun 15;486(2):150-4. doi: 10.1016/j.abb.2009.02.001.

Abstract

The cytoplasmic domain of influenza M2 protein (M2c) consists of 54 amino acid (aa) residues from aa44 to aa97. In this paper, M2c and its deletion mutant M2c(delta47-55) were expressed using prokaryotic expression system. First, glutaraldehyde crosslinking assay showed that M2c had multimerization potential mediated by aa47-55. Then, M2c, instead of M2c(delta47-55), directed eGFP from the whole cell localization to a predominately perinuclear region in CHO cells, which indicated that aa47-55 of M2c mediated the localization. Moreover, M2c colocalized with caveolin-1 (Cav) when CHO cells were cotransfected with Cav. A caveolin-1 binding motif phixxxxphixxphi (phi represents aromatic amino acid residues) in aa47-55 of M2c was found by sequence alignment and analysis. Further overlay ELISA result showed that M2c, but not M2c(delta47-55), bound to prokaryotically expressed cholesterol-free Cav(2-101), which illustrated the interaction could be cholesterol-independent. That was the first report of cellular protein bound to M2c.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Binding Sites
  • Caveolin 1 / chemistry*
  • Caveolin 1 / genetics
  • Caveolin 1 / metabolism*
  • DNA Primers
  • Gene Amplification
  • Influenza A virus / genetics
  • Influenza A virus / metabolism
  • Microscopy, Confocal
  • Models, Molecular
  • Polymerase Chain Reaction
  • Protein Conformation
  • Transfection
  • Viral Matrix Proteins / chemistry*
  • Viral Matrix Proteins / genetics
  • Viral Matrix Proteins / metabolism*
  • Viral Proteins / chemistry
  • Viral Proteins / metabolism

Substances

  • Caveolin 1
  • DNA Primers
  • M2 protein, Influenza A virus
  • Viral Matrix Proteins
  • Viral Proteins