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Fanconi Anaemia E3 Ligase complex activity is regulated by a druggable metabolite binding site in FANCX

Sharp, M. F.; Gee, Y. S.; Luu, J.; Cowley, K.; Beetham, H.; Langendorf, C. G.; Oakhill, J. S.; Scott, J. W.; Cavero, D.; Minguillon, J.; Che, D.; Baell, J. B.; Deans, A. J.; Surralles, J.; Simpson, K. J.; Crismani, W.

2026-08-07 cancer biology
10.64898/2026.08.07.743451 bioRxiv
Show abstract

The Fanconi anaemia (FA) DNA repair pathway is an emerging target for precision cancer therapy. Using a high-throughput FANCD2-monoubiquitination assay, we identified a class of small molecules, including MSG010, that inhibit the FA E3 ligase complex in vitro. Because these molecules, and the metabolite, palmitoyl-CoA, are known to engage allosteric drug and metabolite (ADaM) binding site in AMP-activated kinase (AMPK), we hypothesised that a similar pocket exists within the FA complex. Supporting this, long-chain, but not short-chain, fatty acyl-CoA molecules inhibited the FA E3 ligase complex activity, and sequence analysis revealed similarity between the AMPK ADaM site and a WD40 repeat in the FA subunit FANCX. Targeted mutagenesis of this FANCX region disrupted E3 ligase activity or abolished inhibition by MSG010, suggesting the presence of an ADaM-like site in FANCX. Moreover, MSG010 preferentially killed BRCA1-deficient cells in vitro. These findings identify a putative small-molecule binding site in the FA pathway that may be developed further to test for exploitation as anticancer therapeutics.

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