The AAA ATPase p97/VCP interacts with its alternative co-factors, Ufd1-Npl4 and p47, through a common bipartite binding mechanism

J Biol Chem. 2004 Nov 26;279(48):49609-16. doi: 10.1074/jbc.M408695200. Epub 2004 Sep 15.

Abstract

The AAA ATPase p97/VCP forms complexes with different adapters to fulfill distinct cellular functions. We analyzed the structural organization of the Ufd1-Npl4 adapter complex and its interaction with p97 and compared it with another adapter, p47. We found that the binary Ufd1-Npl4 complex forms a heterodimer that cooperatively interacts with p97 via a bipartite binding mechanism. Binding site 1 (BS1) is a short hydrophobic stretch in the C-terminal domain of Ufd1. The second binding site is located at the N terminus of Npl4 and is activated upon binding of Ufd1 to Npl4. It consists of about 80 amino acids that are predicted to form a ubiquitin fold domain (UBD). Despite the lack of overall homology between Ufd1-Npl4 and p47, both adapters use identical binding mechanisms. Like the ubiquitin fold ubiquitin regulatory X (UBX) domain in p47, the Npl4-UBD interacts with p97 via the loop between its strands 3 and 4 and a conserved arginine in strand 1. Furthermore, we identified a region in p47 homologous to Ufd1-BS1. The UBD/UBX and the BS1 of both adapters interact with p97 independently, whereas homologous binding sites in both adapters compete for binding to p97. In contrast to p47, however, Ufd1-Npl4 does not regulate the ATPase activity of p97; nor does a variant of p47 that contains both binding sites but lacks the N-terminal domains. Therefore, the binding sites alone do not regulate p97 directly but rather serve as anchor points to position adapter-specific domains at critical locations to modulate p97-mediated reactions.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenosine Triphosphatases
  • Animals
  • Binding Sites
  • Carrier Proteins / metabolism*
  • Cell Cycle Proteins / metabolism*
  • Down-Regulation
  • Mutation
  • Nuclear Pore Complex Proteins / genetics
  • Nuclear Pore Complex Proteins / metabolism
  • Nucleocytoplasmic Transport Proteins
  • Protein Binding
  • Protein Structure, Tertiary
  • Proteins / metabolism*
  • Rats
  • Saccharomyces cerevisiae Proteins / genetics
  • Saccharomyces cerevisiae Proteins / metabolism
  • Soluble N-Ethylmaleimide-Sensitive Factor Attachment Proteins
  • Valosin Containing Protein
  • Vesicular Transport Proteins / metabolism*

Substances

  • Carrier Proteins
  • Cell Cycle Proteins
  • NPL4 protein, S cerevisiae
  • Nploc4 protein, rat
  • Nsfl1c protein, rat
  • Nuclear Pore Complex Proteins
  • Nucleocytoplasmic Transport Proteins
  • Proteins
  • Saccharomyces cerevisiae Proteins
  • Soluble N-Ethylmaleimide-Sensitive Factor Attachment Proteins
  • Ufd1 protein, rat
  • Vesicular Transport Proteins
  • Adenosine Triphosphatases
  • CDC48 protein, S cerevisiae
  • Valosin Containing Protein
  • Vcp protein, rat