USP11 mediates repair of DNA–protein cross-links by deubiquitinating SPRTN metalloprotease

• Published in: The Journal of biological chemistry Vol. 296; p. 100396
• Main Authors: Perry, Megan, Biegert, Meghan, Kollala, Sai Sundeep, Mallard, Halle, Su, Grace, Kodavati, Manohar, Kreiling, Natasha, Holbrook, Alexander, Ghosal, Gargi
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01.01.2021; American Society for Biochemistry and Molecular Biology
• Subjects: Cell Line; Cell Line, Tumor; Chromatin - metabolism; Cross-Linking Reagents - chemistry; deubiquitination; DNA Repair; DNA-Binding Proteins - genetics; DNA-Binding Proteins - metabolism; DPC; DPC repair; Genomic Instability; Humans; monoubiquitinated SPRTN; Proteolysis; SPRTN; Thiolester Hydrolases - genetics; Thiolester Hydrolases - metabolism; TOP1-ccs; Ubiquitination; USP11; DNMT1; RADAR; USP11; CPT; Ub-SPRTN; HR; HU; SFB; DUB; DSB; TOP1-ccs; TLS; monoubiquitinated SPRTN; ssDNA; TAP-MS; MMC; SSB; IR; TOP2-ccs; Co-IP; DPC repair; USP; DPC; RJALS; SPRTN; deubiquitination; NER; TOP2; TOP1; dsDNA
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1016/j.jbc.2021.100396

DNA–protein cross-links (DPCs) are toxic DNA lesions that interfere with DNA metabolic processes such as replication, transcription, and recombination. USP11 deubiquitinase participates in DNA repair, but the role of USP11 in DPC repair is not known. SPRTN is a replication-coupled DNA-dependent metalloprotease that cleaves proteins cross-linked to DNA to promote DPC repair. SPRTN function is tightly regulated by a monoubiquitin switch that controls SPRTN auto-proteolysis and chromatin accessibility during DPC repair. Previously, VCPIP1 and USP7 deubiquitinases have been shown to regulate SPRTN. Here, we identify USP11 as an SPRTN deubiquitinase. USP11 interacts with SPRTN and cleaves monoubiquitinated SPRTN in cells and in vitro. USP11 depletion impairs SPRTN deubiquitination and promotes SPRTN auto-proteolysis in response to formaldehyde-induced DPCs. Loss of USP11 causes an accumulation of unrepaired DPCs and cellular hypersensitivity to treatment with DPC-inducing agents. Our findings show that USP11 regulates SPRTN auto-proteolysis and SPRTN-mediated DPC repair to maintain genome stability.