Subquinocin, a small molecule inhibitor of CYLD and USP-family deubiquitinating enzymes, promotes NF-?B signaling.
The tumor suppressor CYLD negatively regulates polyubiquitination-dependent cellular signaling such as nuclear factor (NF)-?B signaling. In addition to CYLD, multiple deubiquitinating enzymes (DUBs) are also involved in the regulation of this signaling pathway, and distinct role of CYLD is yet to be clarified. Here, we identified a small chemical named ... Subquinocin that inhibited the DUB activity of recombinant CYLD using a wheat cell-free protein synthesis and an AlphaScreen technology. In cells, Subquinocin increased the polyubiquitination of NEMO and RIP1 and enhanced NF-?B activation. Modeling and mutation analyses indicated that Subquinocin interacted with Y940 in CYLD, which locates close to catalytic center of CYLD, and is conserved among the USP-family DUBs. Further biochemical evaluation revealed that Subquinocin inhibited USP-family DUBs, but not other family DUBs including OTU. Although Subquinocin showed a broad specificity toward USP-family DUBs, the inhibitory effect of Subquinocin on NF-?B signaling was negligible in CYLD-KO cells, indicating that CYLD is a major target of Subquinocin on the suppression of NF-?B signaling. In conclusion, Subquinocin identified here is a useful tool to analyze the signal transduction mediated by USP-family DUBs.
Mesh Terms:
Amino Acid Sequence, Antineoplastic Agents, Cell Line, Tumor, Deubiquitinating Enzyme CYLD, Drug Screening Assays, Antitumor, Enzyme Inhibitors, Gene Expression Regulation, Genes, Tumor Suppressor, Glutathione Transferase, Humans, Molecular Docking Simulation, Mutation, NF-kappa B, Nuclear Pore Complex Proteins, Protein Binding, Protein Conformation, RNA-Binding Proteins, Recombinant Proteins, Signal Transduction, Tumor Suppressor Proteins, Ubiquitin-Specific Proteases, Ubiquitination
Amino Acid Sequence, Antineoplastic Agents, Cell Line, Tumor, Deubiquitinating Enzyme CYLD, Drug Screening Assays, Antitumor, Enzyme Inhibitors, Gene Expression Regulation, Genes, Tumor Suppressor, Glutathione Transferase, Humans, Molecular Docking Simulation, Mutation, NF-kappa B, Nuclear Pore Complex Proteins, Protein Binding, Protein Conformation, RNA-Binding Proteins, Recombinant Proteins, Signal Transduction, Tumor Suppressor Proteins, Ubiquitin-Specific Proteases, Ubiquitination
Biochem Biophys Res Commun
Date: Dec. 26, 2019
PubMed ID: 31898971
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