title: A selective USP1-UAF1 inhibitor links deubiquitination to DNA damage responses.
abstract: Protein ubiquitination and deubiquitination are central to the control of a large number of cellular pathways and signaling networks in eukaryotes. Although the essential roles of ubiquitination have been established in the eukaryotic DNA damage response, the deubiquitination process remains poorly defined. Chemical probes that perturb the activity of deubiquitinases (DUBs) are needed to characterize the cellular function of deubiquitination. Here we report ML323 (2), a highly potent inhibitor of the USP1-UAF1 deubiquitinase complex with excellent selectivity against human DUBs, deSUMOylase, deneddylase and unrelated proteases. Using ML323, we interrogated deubiquitination in the cellular response to UV- and cisplatin-induced DNA damage and revealed new insights into the requirement of deubiquitination in the DNA translesion synthesis and Fanconi anemia pathways. Moreover, ML323 potentiates cisplatin cytotoxicity in non-small cell lung cancer and osteosarcoma cells. Our findings point to USP1-UAF1 as a key regulator of the DNA damage response and a target for overcoming resistance to the platinum-based anticancer drugs.
authors: Department of Chemistry and Biochemistry, University of Delaware, Newark, Delaware, USA.
authors: Qin
authors: Q
authors: Liang
authors: National Center for Advancing Translational Sciences, National Institutes of Health, Bethesda, Maryland, USA.
authors: Thomas S
authors: TS
authors: Dexheimer
authors: Department of Chemistry and Biochemistry, University of Delaware, Newark, Delaware, USA.
authors: Ping
authors: P
authors: Zhang
authors: National Center for Advancing Translational Sciences, National Institutes of Health, Bethesda, Maryland, USA.
authors: Andrew S
authors: AS
authors: Rosenthal
authors: Department of Chemistry and Biochemistry, University of Delaware, Newark, Delaware, USA.
authors: Mark A
authors: MA
authors: Villamil
authors: Department of Chemistry, University of California-Riverside, Riverside, California, USA.
authors: Changjun
authors: C
authors: You
authors: State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang, Jiangxi, China.
authors: Qiuting
authors: Q
authors: Zhang
authors: Department of Chemistry and Biochemistry, University of Delaware, Newark, Delaware, USA.
authors: Junjun
authors: J
authors: Chen
authors: Department of Chemistry and Biochemistry, University of Delaware, Newark, Delaware, USA.
authors: Christine A
authors: CA
authors: Ott
authors: National Center for Advancing Translational Sciences, National Institutes of Health, Bethesda, Maryland, USA.
authors: Hongmao
authors: H
authors: Sun
authors: National Center for Advancing Translational Sciences, National Institutes of Health, Bethesda, Maryland, USA.
authors: Diane K
authors: DK
authors: Luci
authors: Department of Chemistry, University of California-Riverside, Riverside, California, USA.
authors: Bifeng
authors: B
authors: Yuan
authors: National Center for Advancing Translational Sciences, National Institutes of Health, Bethesda, Maryland, USA.
authors: Anton
authors: A
authors: Simeonov
authors: National Center for Advancing Translational Sciences, National Institutes of Health, Bethesda, Maryland, USA.
authors: Ajit
authors: A
authors: Jadhav
authors: Laboratory of Macromolecular Analysis and Proteomics, Albert Einstein College of Medicine, Bronx, New York, USA.
authors: Hui
authors: H
authors: Xiao
authors: Department of Chemistry, University of California-Riverside, Riverside, California, USA.
authors: Yinsheng
authors: Y
authors: Wang
authors: National Center for Advancing Translational Sciences, National Institutes of Health, Bethesda, Maryland, USA.
authors: David J
authors: DJ
authors: Maloney
authors: Department of Chemistry and Biochemistry, University of Delaware, Newark, Delaware, USA.
authors: Zhihao
authors: Z
authors: Zhuang
mesh_terms: D000465:Algorithms; D000970:Antineoplastic Agents / Q000138:chemical synthesis* / Q000494:pharmacology*; D029681:Arabidopsis Proteins / Q000037:antagonists & inhibitors*; D002087:Butyrates / Q000494:pharmacology; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D002945:Cisplatin / Q000494:pharmacology; D003114:Colony-Forming Units Assay; D004249:DNA Damage / Q000235:genetics / Q000502:physiology*; D004273:DNA, Neoplasm / Q000037:antagonists & inhibitors / Q000096:biosynthesis; D019008:Drug Resistance, Neoplasm; D004591:Electrophoresis, Polyacrylamide Gel; D005199:Fanconi Anemia / Q000235:genetics; D052236:Fanconi Anemia Complementation Group D2 Protein / Q000037:antagonists & inhibitors; D057166:High-Throughput Screening Assays; D006801:Humans; D007202:Indicators and Reagents; D009687:Nuclear Proteins / Q000037:antagonists & inhibitors*; D010671:Phenylurea Compounds / Q000494:pharmacology; D010868:Pimozide / Q000494:pharmacology; D018809:Proliferating Cell Nuclear Antigen / Q000187:drug effects / Q000378:metabolism; D034741:RNA, Small Interfering / Q000235:genetics; D011994:Recombinant Proteins / Q000737:chemistry; D011995:Recombination, Genetic / Q000187:drug effects; D012854:Sister Chromatid Exchange / Q000187:drug effects; D064570:Ubiquitin-Specific Proteases / Q000037:antagonists & inhibitors*; D054875:Ubiquitination / Q000187:drug effects*
chemical_list: D000970:Antineoplastic Agents; D029681:Arabidopsis Proteins; D002087:Butyrates; D002945:Cisplatin; D004273:DNA, Neoplasm; D052236:Fanconi Anemia Complementation Group D2 Protein; D007202:Indicators and Reagents; D009687:Nuclear Proteins; D010671:Phenylurea Compounds; D010868:Pimozide; D018809:Proliferating Cell Nuclear Antigen; D034741:RNA, Small Interfering; D011994:Recombinant Proteins; D064570:Ubiquitin-Specific Proteases; C499905:FANCD2 protein, human; C453899:GW 7647; C526165:WDR48 protein, human; C579228:USP1 protein, human