Identification of ubiquitin-like protein-binding subunits of the 26S proteasome

Ubiquitin-like proteins Rad23 and Dsk2 have recently been shown to be capable of binding both polyubiquitin chains and the 26S proteasome. The ubiquitin-like domains (Ubls) of Rad23 and Dsk2 are indispensable for their interaction with the 26S proteasome, but the proteasome subunits capable of bindi...

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Published inBiochemical and biophysical research communications Vol. 296; no. 4; pp. 813 - 819
Main Authors Saeki, Yasushi, Sone, Takayuki, Toh-e, Akio, Yokosawa, Hideyoshi
Format Journal Article
LanguageEnglish
Published United States Elsevier Inc 30.08.2002
Subjects
Binding, Competitive
Blotting, Western
Cell Cycle Proteins
Cross-Linking Reagents - pharmacology
DNA-Binding Proteins - chemistry
Dsk2
Fungal Proteins - chemistry
Fungal Proteins - metabolism
Glutathione Transferase - metabolism
Open Reading Frames
Peptide Hydrolases - chemistry
Plasmids - metabolism
Precipitin Tests
Proteasome
Proteasome Endopeptidase Complex
Protein Binding
Proteins - chemistry
Rad23
Recombinant Fusion Proteins - metabolism
Recombinant Proteins - metabolism
Saccharomyces cerevisiae - metabolism
Saccharomyces cerevisiae Proteins
Ubiquitin
Ubiquitin - metabolism
Ubiquitin-like domain
Ubiquitins - chemistry
Ubiquitins - metabolism
Ubiquitin
Ubiquitin-like domain
Dsk2
Rad23
Proteasome
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Summary:Ubiquitin-like proteins Rad23 and Dsk2 have recently been shown to be capable of binding both polyubiquitin chains and the 26S proteasome. The ubiquitin-like domains (Ubls) of Rad23 and Dsk2 are indispensable for their interaction with the 26S proteasome, but the proteasome subunits capable of binding the Ubl have not been identified. Here, we report that the Ubls of both Rad23 and Dsk2 can bind with the 19S regulatory particle (RP) of the 26S proteasome in vivo and in vitro. A competition assay using the respective Ubls of Rad23 and Dsk2 revealed that they bind to the RP in a competitive manner. The base subcomplex of the RP was found to have the ability to bind the Ubl. By cross-linking experiments, Rpn1 and Rpn2 were identified as Ubl-binding subunits. Taken together, the results suggest that the Rpn1 and Rpn2 in the base subcomplex form the receptor for the ubiquitin-like protein.
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ISSN:0006-291X
1090-2104
DOI:10.1016/S0006-291X(02)02002-8