Components of the ubiquitin-proteasome pathway compete for surfaces on Rad23 family proteins.
BACKGROUND
The delivery of ubiquitinated proteins to the proteasome for degradation is a key step in the regulation of the ubiquitin-proteasome pathway, yet the mechanisms underlying this step are not understood in detail. The Rad23 family of proteins is known to bind ubiquitinated proteins through its two ubiquitin-associated (UBA) domains, and may participate in the delivery of ubiquitinated proteins to the proteasome through docking via the Rad23 ubiquitin-like (UBL) domain.


RESULTS
In this study, we investigate how the interaction between the UBL and UBA domains may modulate ubiquitin recognition and the delivery of ubiquitinated proteins to the proteasome by autoinhibition. We have explored a competitive binding model using specific mutations in the UBL domain. Disrupting the intramolecular UBL-UBA domain interactions in HHR23A indeed potentiates ubiquitin-binding. Additionally, the analogous surface on the Rad23 UBL domain overlaps with that required for interaction with both proteasomes and the ubiquitin ligase Ufd2. We have found that mutation of residues on this surface affects the ability of Rad23 to deliver ubiquitinated proteins to the proteasome.


CONCLUSIONS
We conclude that the competition of ubiquitin-proteasome pathway components for surfaces on Rad23 is important for the role of the Rad23 family proteins in proteasomal targeting.
Goh
Amanda M
AM
Department of Pathology, Harvard Medical School, Boston, Massachusetts, USA. amgoh@imcb.a-star.edu.sg
Walters
Kylie J
KJ
Elsasser
Suzanne
S
Verma
Rati
R
Deshaies
Raymond J
RJ
Finley
Daniel
D
Howley
Peter M
PM
D001667:Binding, Competitive / Q000235:genetics; D045643:DNA Repair Enzymes / Q000737:chemistry* / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000737:chemistry* / Q000378:metabolism*; D006801:Humans; D008954:Models, Biological; D009154:Mutation; D025821:Polyubiquitin / Q000378:metabolism; D046988:Proteasome Endopeptidase Complex / Q000378:metabolism*; D017434:Protein Structure, Tertiary / Q000235:genetics; D025801:Ubiquitin / Q000235:genetics / Q000378:metabolism*; D044767:Ubiquitin-Protein Ligases / Q000378:metabolism; D054875:Ubiquitination / Q000235:genetics
D004268:DNA-Binding Proteins; D025801:Ubiquitin; D025821:Polyubiquitin; C086985:RAD23A protein, human; D044767:Ubiquitin-Protein Ligases; D046988:Proteasome Endopeptidase Complex; D045643:DNA Repair Enzymes