Subunit interaction maps for the regulatory particle of the 26S proteasome and the COP9 signalosome.
The 26S proteasome plays a major role in eukaryotic protein breakdown, especially for ubiquitin-tagged proteins. Substrate specificity is conferred by the regulatory particle (RP), which can dissociate into stable lid and base subcomplexes. To help define the molecular organization of the RP, we tested all possible paired interactions among subunits from Saccharomyces cerevisiae by yeast two-hybrid analysis. Within the base, a Rpt4/5/3/6 interaction cluster was evident. Within the lid, a structural cluster formed around Rpn5/11/9/8. Interactions were detected among synonymous subunits (Csn4/5/7/6) from the evolutionarily related COP9 signalosome (CSN) from Arabidopsis, implying a similar quaternary arrangement. No paired interactions were detected between lid, base or core particle subcomplexes, suggesting that stable contacts between them require prior assembly. Mutational analysis defined the ATPase, coiled-coil, PCI and MPN domains as important for RP assembly. A single residue in the vWA domain of Rpn10 is essential for amino acid analog resistance, for degrading a ubiquitin fusion degradation substrate and for stabilizing lid-base association. Comprehensive subunit interaction maps for the 26S proteasome and CSN support the ancestral relationship of these two complexes.
Fu
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H
Institute of Botany, Academia Sinica, 128, Sec 2, Academy Road, Taipei, Taiwan 115, Republic of China. hongyong@gate.sinica.edu.tw
Reis
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N
Lee
Y
Y
Glickman
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MH
Vierstra
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RD
D029681:Arabidopsis Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D001224:Aspartic Acid / Q000378:metabolism; D000075686:COP9 Signalosome Complex; D003546:Cysteine Endopeptidases / Q000737:chemistry / Q000378:metabolism*; D006868:Hydrolysis; D009097:Multienzyme Complexes / Q000737:chemistry / Q000378:metabolism*; D046912:Multiprotein Complexes; D016296:Mutagenesis; D010447:Peptide Hydrolases; D046988:Proteasome Endopeptidase Complex; D011485:Protein Binding; D011506:Proteins / Q000737:chemistry / Q000378:metabolism*; D015398:Signal Transduction; D013379:Substrate Specificity
D029681:Arabidopsis Proteins; D009097:Multienzyme Complexes; D046912:Multiprotein Complexes; D011506:Proteins; D001224:Aspartic Acid; D010447:Peptide Hydrolases; D000075686:COP9 Signalosome Complex; D003546:Cysteine Endopeptidases; D046988:Proteasome Endopeptidase Complex