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Deubiquitination of FANCD2, FANCI, and PCNA by USP1 is essential for DNA repair signalling. Reconstitution of the system reveals that USP1-mediated specificity towards K561 of FANCD2 is directed by a unique sequence at USP1's N-terminus.

Specificity for deubiquitination of monoubiquitinated FANCD2 is driven by the N-terminus of USP1