Structural basis of FANCD2 deubiquitination by USP1−UAF1

Martin L. Rennie (Lead / Corresponding author), Connor Arkinson, Viduth K. Chaugule, Rachel Toth, Helen Walden (Lead / Corresponding author)

Research output: Contribution to journalArticlepeer-review

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.
Original languageEnglish
Number of pages24
JournalNature Structural & Molecular Biology
Early online date1 Apr 2021
DOIs
Publication statusE-pub ahead of print - 1 Apr 2021

Keywords

  • ubiquitin
  • specificity
  • DNA repair
  • Fanconi Anemia
  • cryo-EM

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