1. Structural basis of FANCD2 deubiquitination by USP1-UAF1.
- Author
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Rennie ML, Arkinson C, Chaugule VK, Toth R, and Walden H
- Subjects
- DNA Damage genetics, DNA Repair genetics, Deubiquitinating Enzymes chemistry, Deubiquitinating Enzymes genetics, Deubiquitinating Enzymes ultrastructure, Fanconi Anemia genetics, Fanconi Anemia pathology, Fanconi Anemia Complementation Group D2 Protein genetics, HeLa Cells, Humans, Nuclear Proteins genetics, Protein Binding genetics, Protein Conformation, Ubiquitin-Specific Proteases genetics, Ubiquitination genetics, Fanconi Anemia Complementation Group D2 Protein ultrastructure, Nuclear Proteins ultrastructure, Ubiquitin-Specific Proteases ultrastructure
- Abstract
Ubiquitin-specific protease 1 (USP1) acts together with the cofactor UAF1 during DNA repair processes to specifically remove monoubiquitin signals. One substrate of the USP1-UAF1 complex is the monoubiquitinated FANCI-FANCD2 heterodimer, which is involved in the repair of DNA interstrand crosslinks via the Fanconi anemia pathway. Here we determine structures of human USP1-UAF1 with and without ubiquitin and bound to monoubiquitinated FANCI-FANCD2. The crystal structures of USP1-UAF1 reveal plasticity in USP1 and key differences to USP12-UAF1 and USP46-UAF1, two related proteases. A cryo-EM reconstruction of USP1-UAF1 in complex with monoubiquitinated FANCI-FANCD2 highlights a highly orchestrated deubiquitination process, with USP1-UAF1 driving conformational changes in the substrate. An extensive interface between UAF1 and FANCI, confirmed by mutagenesis and biochemical assays, provides a molecular explanation for the requirement of both proteins, despite neither being directly involved in catalysis. Overall, our data provide molecular details of USP1-UAF1 regulation and substrate recognition.
- Published
- 2021
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