Structural and functional characterization of Rpn12 identifies residues required for Rpn10 proteasome incorporation.
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.
Department of Biochemistry, University of Oxford, Oxford OX1 3QU, UK.
Jonas
J
Boehringer
Christiane
C
Riedinger
Konstantinos
K
Paraskevopoulos
Eachan O D
EO
Johnson
Edward D
ED
Lowe
Christina
C
Khoudian
Dominique
D
Smith
Martin E M
ME
Noble
Colin
C
Gordon
Jane A
JA
Endicott
D000818:Animals; D029681:Arabidopsis Proteins / Q000737:chemistry; D000075686:COP9 Signalosome Complex; D002352:Carrier Proteins / Q000737:chemistry / Q000378:metabolism*; D002942:Circular Dichroism; D018360:Crystallography, X-Ray; D029721:Drosophila Proteins / Q000737:chemistry; D008869:Microtubule-Associated Proteins / Q000737:chemistry; D008958:Models, Molecular; D016297:Mutagenesis, Site-Directed; D010446:Peptide Fragments / Q000737:chemistry / Q000378:metabolism; D046988:Proteasome Endopeptidase Complex / Q000378:metabolism*; D011485:Protein Binding; D011487:Protein Conformation; D025941:Protein Interaction Mapping; D017434:Protein Structure, Tertiary; D016601:RNA-Binding Proteins; D011994:Recombinant Proteins / Q000378:metabolism; D012568:Schizosaccharomyces / Q000235:genetics / Q000378:metabolism; D029702:Schizosaccharomyces pombe Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D013329:Structure-Activity Relationship; D025801:Ubiquitin / Q000378:metabolism; D051540:Winged-Helix Transcription Factors / Q000737:chemistry
D029681:Arabidopsis Proteins; C535125:CSN7 protein, Arabidopsis; D002352:Carrier Proteins; D029721:Drosophila Proteins; C468124:EIF3K protein, human; D008869:Microtubule-Associated Proteins; D010446:Peptide Fragments; D016601:RNA-Binding Proteins; C098743:RPN12 protein, S pombe; D011994:Recombinant Proteins; C470518:Rpn6 protein, Drosophila; D029702:Schizosaccharomyces pombe Proteins; D025801:Ubiquitin; D051540:Winged-Helix Transcription Factors; C498507:pus1 protein, S pombe; D000075686:COP9 Signalosome Complex; D046988:Proteasome Endopeptidase Complex