title: The role of USP1 autocleavage in DNA interstrand crosslink repair.
abstract: The Fanconi anemia (FA) pathway regulates DNA interstrand crosslink (ICL) repair. A critical step in this pathway is mono-ubiquitination of FANCD2 (FANCD2-Ub). Deubiquitinase USP1 removes ubiquitin from FANCD2 resulting in inactivation of the FA pathway. USP1 is autocleaved and subsequently degraded for its down-regulation. Here, we investigated the functional consequences of Usp1-autocleavage defect. Usp1-autocleavage mutant (Usp1(GG/AA) ) corrected the level of Fancd2-Ub similar to Usp1(WT) in Usp1(-/-) MEFs. However, Usp1(GG/AA) only partially rescued MMC sensitivity with defective Fancd2 foci formation and homologous recombination defect. Contrary to this, Usp1(GG/AA) was capable of recovering UV resistance of Usp1(-/-) MEFs to a similar extent with Usp1(WT) . Taken together, our findings suggest that Usp1 regulation by autocleavage is critical for Usp1 to exert its function in ICL repair.
authors.affiliation: Department of Pharmacology, Medical Research Center for Gene Regulation, Chonnam National University Medical School, Gwangju, Korea.
authors.forename: Mira
authors.initials: M
authors.lastname: Kim
authors.affiliation: Department of Pharmacology, Medical Research Center for Gene Regulation, Chonnam National University Medical School, Gwangju, Korea.
authors.forename: Jung Min
authors.initials: JM
authors.lastname: Kim
mesh_terms: D019943:Amino Acid Substitution; D000818:Animals; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects / Q000528:radiation effects; D003432:Cross-Linking Reagents / Q000633:toxicity; D004247:DNA / Q000187:drug effects / Q000528:radiation effects; D004260:DNA Repair; D052236:Fanconi Anemia Complementation Group D2 Protein / Q000378:metabolism*; D055879:Gene Knock-In Techniques; D055786:Gene Knockout Techniques; D051379:Mice; D016297:Mutagenesis, Site-Directed; D009154:Mutation; D041681:NIH 3T3 Cells; D018809:Proliferating Cell Nuclear Antigen / Q000378:metabolism*; D055550:Protein Stability; D059748:Proteolysis; D011838:Radiation-Sensitizing Agents; D011994:Recombinant Proteins / Q000378:metabolism; D064570:Ubiquitin-Specific Proteases / Q000235:genetics / Q000378:metabolism*; D054875:Ubiquitination; D014466:Ultraviolet Rays / Q000009:adverse effects
chemical_list: D003432:Cross-Linking Reagents; C499904:Fancd2 protein, mouse; D052236:Fanconi Anemia Complementation Group D2 Protein; D018809:Proliferating Cell Nuclear Antigen; D011838:Radiation-Sensitizing Agents; D011994:Recombinant Proteins; D004247:DNA; D064570:Ubiquitin-Specific Proteases; C579227:Usp1 protein, mouse
keywords: DNA interstand crosslink repair; Fancd2; PCNA; Usp1; autocleavage; deubiquitinating enzyme