The Fanconi anemia core complex promotes CtIP-dependent end-resection to drive homologous recombination at DNA double-strand breaks

Bert van de Kooij*, Fenna J van der Wal, Magdalena B Rother, Pau Creixell, Merula Stout, Wouter Wiegant, Brian A Joughin, Julia Vornberger, Marcel A T M van Vugt, Matthias Altmeyer, Michael B Yaffe, Haico van Attikum

*Corresponding author voor dit werk

    Onderzoeksoutput: VoordrukAcademic

    Samenvatting

    Homologous Recombination (HR) is a high-fidelity repair mechanism of DNA Double-Strand Breaks (DSBs), which are induced by irradiation, genotoxic chemicals or physiological DNA damaging processes. DSBs are also generated as intermediates during the repair of interstrand crosslinks (ICLs). In this context, the Fanconi anemia (FA) core complex, which is effectively recruited to ICLs, promotes HR-mediated DSB-repair. However, whether the FA core complex also promotes HR at ICL-independent DSBs remains controversial. Here, we identified the FA core complex members FANCL and Ube2T as HR-promoting factors in a CRISPR/Cas9-based screen with cells carrying the DSB-repair reporter DSB-Spectrum. Using isogenic cell-line models, we validated the HR-function of FANCL and Ube2T, and demonstrated a similar function for their ubiquitination-substrate FANCD2. We further show that FANCL and Ube2T are directly recruited to DSBs and are required for the accumulation of FANCD2 at these break sites. Mechanistically, we demonstrate that FANCL ubiquitin ligase activity is required for the accumulation of the nuclease CtIP at DSBs, and consequently for optimal end-resection and Rad51 loading. CtIP overexpression rescues HR in FANCL-deficient cells, validating that FANCL primarily regulates HR by promoting CtIP recruitment. Together, these data demonstrate that the FA core complex and FANCD2 have a dual genome maintenance function by promoting repair of DSBs as well as the repair of ICLs.

    Originele taal-2English
    UitgeverBioRxiv
    Aantal pagina's40
    DOI's
    StatusPublished - 6-sep.-2023

    Publicatie series

    NaambioRxiv
    UitgeverijCold Spring Harbor Labs Journals
    ISSN van geprinte versie2692-8205

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