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bioRxiv · 10.1101/2020.12.05.412924

Structural basis of FANCD2 deubiquitination by USP1-UAF1

Rennie, M. L.·Arkinson, C.·Chaugule, V. K.·Toth, R.·Walden, H.
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Abstract

Ubiquitin-Specific Protease 1 (USP1), together with the cofactor UAF1, acts during DNA repair processes to specifically to remove mono-ubiquitin signals. The mono-ubiquitinated FANCI-FANCD2 heterodimer is one such substrate and 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 mono-ubiquitinated FANCI-FANCD2 substrate. Crystal structures of USP1-UAF1 reveal plasticity in USP1 and key differences to USP12-UAF1 and USP46-UAF1. A cryoEM reconstruction of USP1-UAF1 in complex mono-ubiquitinated 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 their requirement despite neither being directly involved in catalysis. Overall, our data provide molecular details of USP1-UAF1 regulation and substrate recognition.

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Rennie, M. L., Arkinson, C., Chaugule, V. K., Toth, R., Walden, H.. 2020-12-06. Structural basis of FANCD2 deubiquitination by USP1-UAF1. https://doi.org/10.1101/2020.12.05.412924

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