Adenovirus protein involved in virus internalization recruits ubiquitin-protein ligases

Biochemistry. 2002 Dec 3;41(48):14299-305. doi: 10.1021/bi020125b.

Abstract

Adenovirus penton base protein is involved in virus internalization. Searching for the cellular partners of this protein, we used dodecahedra, adenovirus subviral particles composed of 12 bases, for screening a human lung expression library. This screen yielded three ubiquitin-protein ligases, WWP1, WWP2, and AIP4, all of which belong to the HECT family and contain multiple WW domains. The xPPxY motif, known to interact with WW domains in partner proteins occurs twice in the N-terminal part of the base polypeptide chain. The recruitment of three ubiquitin-protein ligases was shown for two distinct virus serotypes, Ad2 and Ad3. The first N-terminal xPPxY motif in the base protein sequence is indispensable for the interaction. The association in vitro was shown by the protein overlay technique and in vivo by cotransfection followed by immunoprecipitation. The binding parameters studied by surface plasmon resonance confirmed the interaction of base protein with three ubiquitin-protein ligases. In case of WWP1 when the saturation of binding was achieved, the apparent dissociation constant of 65nM was calculated. This is the first demonstration of the interaction of nonenveloped viruses with ubiquitin-protein ligases of host cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenoviruses, Human / physiology*
  • Amino Acid Motifs / genetics
  • Amino Acid Sequence
  • Baculoviridae / genetics
  • Calcium-Binding Proteins / biosynthesis
  • Calcium-Binding Proteins / genetics
  • Calcium-Binding Proteins / metabolism*
  • Capsid / metabolism
  • Capsid / physiology*
  • Capsid Proteins*
  • Endocytosis* / genetics
  • Gene Expression Regulation, Viral
  • Gene Library
  • HeLa Cells
  • Humans
  • Ligases / biosynthesis
  • Ligases / genetics
  • Ligases / metabolism*
  • Molecular Sequence Data
  • Protein Structure, Tertiary / genetics
  • Repressor Proteins / biosynthesis
  • Repressor Proteins / genetics
  • Repressor Proteins / metabolism*
  • Ubiquitin-Protein Ligases

Substances

  • Calcium-Binding Proteins
  • Capsid Proteins
  • Repressor Proteins
  • penton protein, adenovirus
  • ITCH protein, human
  • WWP1 protein, human
  • WWP2 protein, human
  • Ubiquitin-Protein Ligases
  • Ligases