Structural and biochemical basis of interdependent FANCI-FANCD2 ubiquitination

Lemonidis, K. , Rennie, M. L. , Arkinson, C., Chaugule, V. K. , Clarke, M. , Streetley, J. and Walden, H. (2023) Structural and biochemical basis of interdependent FANCI-FANCD2 ubiquitination. EMBO Journal, 42(3), e111898. (doi: 10.15252/embj.2022111898) (PMID:36385258) (PMCID:PMC9890228)

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Di-monoubiquitination of the FANCI-FANCD2 (ID2) complex is a central and crucial step for the repair of DNA interstrand crosslinks via the Fanconi anaemia pathway. While FANCD2 ubiquitination precedes FANCI ubiquitination, FANCD2 is also deubiquitinated at a faster rate than FANCI, which can result in a FANCI-ubiquitinated ID2 complex (IUbD2). Here, we present a 4.1 Å cryo-EM structure of IUbD2 complex bound to double-stranded DNA. We show that this complex, like ID2Ub and IUbD2Ub, is also in the closed ID2 conformation and clamps on DNA. The target lysine of FANCD2 (K561) becomes fully exposed in the IUbD2-DNA structure and is thus primed for ubiquitination. Similarly, FANCI's target lysine (K523) is also primed for ubiquitination in the ID2Ub-DNA complex. The IUbD2-DNA complex exhibits deubiquitination resistance, conferred by the presence of DNA and FANCD2. ID2Ub-DNA, on the other hand, can be efficiently deubiquitinated by USP1-UAF1, unless further ubiquitination on FANCI occurs. Therefore, FANCI ubiquitination effectively maintains FANCD2 ubiquitination in two ways: it prevents excessive FANCD2 deubiquitination within an IUbD2Ub-DNA complex, and it enables re-ubiquitination of FANCD2 within a transient, closed-on-DNA, IUbD2 complex.

Item Type:Articles
Additional Information:This work was supported by the European Research Council (ERC-2015-CoG-681582 ICLUb) consolidator grant to HW. The authors acknowledge the University of Glasgow (college of MVLS) Structural Biology and Biophysical Characterisation Facility for access to Monolith NanoTemper instrument, used for PIFE experiments.
Glasgow Author(s) Enlighten ID:Clarke, Dr Mairi and Chaugule, Dr Viduth and Lemonidis, Dr Kimon and Arkinson, Connor and Rennie, Dr Martin and Walden, Professor Helen and Streetley, Dr James
Authors: Lemonidis, K., Rennie, M. L., Arkinson, C., Chaugule, V. K., Clarke, M., Streetley, J., and Walden, H.
College/School:College of Medical Veterinary and Life Sciences > School of Infection & Immunity > Centre for Virus Research
College of Medical Veterinary and Life Sciences > School of Molecular Biosciences
Journal Name:EMBO Journal
Publisher:EMBO Press
ISSN (Online):1460-2075
Published Online:17 November 2022
Copyright Holders:Copyright © 2022 The Authors
First Published:First published in EMBO Journal 42(3): e111898
Publisher Policy:Reproduced under a Creative Commons License
Data DOI:10.5525/gla.researchdata.1358

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Project CodeAward NoProject NamePrincipal InvestigatorFunder's NameFunder RefLead Dept
301445The Scottish Macromolecular Imaging Centre (SMIC)David BhellaMedical Research Council (MRC)MC_PC_17135III - Centre for Virus Research
301474The purchase and maintenance of a cryo-electron microscopeDavid BhellaScottish Funding Council (SFC)H17007III - Centre for Virus Research
302391Regulation of DNA interstrand crosslink repair by ubiquitin.Helen WaldenEuropean Research Council (ERC)681506Institute of Molecular, Cell & Systems Biology