Structural and functional characterization of Rpn12 identifies residues required for Rpn10 proteasome incorporation

Biochem J. 2012 Nov 15;448(1):55-65. doi: 10.1042/BJ20120542.

Abstract

The ubiquitin-proteasome system targets selected proteins for degradation by the 26S proteasome. Rpn12 is an essential component of the 19S regulatory particle and plays a role in recruiting the extrinsic ubiquitin receptor Rpn10. In the present paper we report the crystal structure of Rpn12, a proteasomal PCI-domain-containing protein. The structure helps to define a core structural motif for the PCI domain and identifies potential sites through which Rpn12 might form protein-protein interactions. We demonstrate that mutating residues at one of these sites impairs Rpn12 binding to Rpn10 in vitro and reduces Rpn10 incorporation into proteasomes in vivo.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Arabidopsis Proteins / chemistry
  • COP9 Signalosome Complex
  • Carrier Proteins / chemistry
  • Carrier Proteins / metabolism*
  • Circular Dichroism
  • Crystallography, X-Ray
  • Drosophila Proteins / chemistry
  • Microtubule-Associated Proteins / chemistry
  • Models, Molecular
  • Mutagenesis, Site-Directed
  • Peptide Fragments / chemistry
  • Peptide Fragments / metabolism
  • Proteasome Endopeptidase Complex / metabolism*
  • Protein Binding
  • Protein Conformation
  • Protein Interaction Mapping
  • Protein Structure, Tertiary
  • RNA-Binding Proteins
  • Recombinant Proteins / metabolism
  • Schizosaccharomyces / genetics
  • Schizosaccharomyces / metabolism
  • Schizosaccharomyces pombe Proteins / chemistry*
  • Schizosaccharomyces pombe Proteins / genetics
  • Schizosaccharomyces pombe Proteins / metabolism*
  • Structure-Activity Relationship
  • Ubiquitin / metabolism
  • Winged-Helix Transcription Factors / chemistry

Substances

  • Arabidopsis Proteins
  • CSN7 protein, Arabidopsis
  • Carrier Proteins
  • Drosophila Proteins
  • EIF3K protein, human
  • Microtubule-Associated Proteins
  • Peptide Fragments
  • RNA-Binding Proteins
  • RPN12 protein, S pombe
  • Recombinant Proteins
  • Rpn6 protein, Drosophila
  • Schizosaccharomyces pombe Proteins
  • Ubiquitin
  • Winged-Helix Transcription Factors
  • pus1 protein, S pombe
  • COP9 Signalosome Complex
  • Proteasome Endopeptidase Complex