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
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Comparative Study
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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Arabidopsis Proteins / chemistry
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COP9 Signalosome Complex
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Carrier Proteins / chemistry
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Carrier Proteins / metabolism*
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Circular Dichroism
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Crystallography, X-Ray
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Drosophila Proteins / chemistry
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Microtubule-Associated Proteins / chemistry
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Models, Molecular
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Mutagenesis, Site-Directed
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Peptide Fragments / chemistry
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Peptide Fragments / metabolism
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Proteasome Endopeptidase Complex / metabolism*
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Protein Binding
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Protein Conformation
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Protein Interaction Mapping
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Protein Structure, Tertiary
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RNA-Binding Proteins
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Recombinant Proteins / metabolism
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Schizosaccharomyces / genetics
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Schizosaccharomyces / metabolism
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Schizosaccharomyces pombe Proteins / chemistry*
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Schizosaccharomyces pombe Proteins / genetics
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Schizosaccharomyces pombe Proteins / metabolism*
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Structure-Activity Relationship
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Ubiquitin / metabolism
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Winged-Helix Transcription Factors / chemistry
Substances
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Arabidopsis Proteins
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CSN7 protein, Arabidopsis
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Carrier Proteins
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Drosophila Proteins
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EIF3K protein, human
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Microtubule-Associated Proteins
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Peptide Fragments
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RNA-Binding Proteins
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RPN12 protein, S pombe
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Recombinant Proteins
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Rpn6 protein, Drosophila
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Schizosaccharomyces pombe Proteins
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Ubiquitin
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Winged-Helix Transcription Factors
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pus1 protein, S pombe
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COP9 Signalosome Complex
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Proteasome Endopeptidase Complex