title: Ubiquitin-interacting motifs confer full catalytic activity, but not ubiquitin chain substrate specificity, to deubiquitinating enzyme USP37.
abstract: Ubiquitin-specific proteases (USPs) consist of a family of deubiquitinating enzymes with more than 50 members in humans. Three of them, including USP37, contain ubiquitin-interacting motifs (UIMs), an ∼20-amino acid α-helical stretch that binds to ubiquitin. However, the roles of the UIMs in these USP enzymes remain unknown. USP37 has three UIMs, designated here as UIMs 1, 2, and 3 from the N-terminal side, between the Cys and His boxes comprising the catalytic core. Here, we examined the role of the UIMs in USP37 using its mutants that harbor mutations in the UIMs. The nuclear localization of USP37 was not affected by the UIM mutations. However, mutations in UIM2 or UIM3, but not UIM1, resulted in a significant decrease in USP37 binding to ubiquitinated proteins in the cell. In vitro, a region of USP37 harboring the three UIMs also bound to both Lys(48)-linked and Lys(63)-linked ubiquitin chains in a UIM2- and UIM3-dependent manner. The level of USP37 ubiquitination was also reduced by mutations in UIM2 or UIM3, suggesting their role in ubiquitination of USP37 itself. Finally, mutants lacking functional UIM2 or UIM3 exhibited a reduced isopeptidase activity toward ubiquitinated proteins in the cell and both Lys(48)-linked and Lys(63)-linked ubiquitin chains. These results suggested that the UIMs in USP37 contribute to the full enzymatic activity, but not ubiquitin chain substrate specificity, of USP37 possibly by holding the ubiquitin chain substrate in the proximity of the catalytic core.
authors: From the Department of Biological Sciences, Tokyo Institute of Technology, Yokohama 226-8501, Japan.
authors: Hidetaka
authors: H
authors: Tanno
authors: Takeshi
authors: T
authors: Shigematsu
authors: Shuhei
authors: S
authors: Nishikawa
authors: Akira
authors: A
authors: Hayakawa
authors: Kimitoshi
authors: K
authors: Denda
authors: Toshiaki
authors: T
authors: Tanaka
authors: Masayuki
authors: M
authors: Komada
mesh_terms: D020816:Amino Acid Motifs; D000818:Animals; D019556:COS Cells; D002384:Catalysis; D002522:Chlorocebus aethiops; D010450:Endopeptidases / Q000235:genetics / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D009154:Mutation; D025821:Polyubiquitin / Q000235:genetics / Q000378:metabolism*; D013379:Substrate Specificity / Q000502:physiology; D057149:Ubiquitinated Proteins / Q000235:genetics / Q000378:metabolism*; D054875:Ubiquitination / Q000502:physiology*
chemical_list: D057149:Ubiquitinated Proteins; D025821:Polyubiquitin; D010450:Endopeptidases; C559980:USP37 protein, human
keywords: Deubiquitination; Protease; Protein Degradation; Ubiquitin; Ubiquitination