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. Author manuscript; available in PMC: 2020 Nov 3.
Published in final edited form as: Nature. 2020 Mar 4;579(7800):603–608. doi: 10.1038/s41586-020-2059-5

Fig. 1. AA-ICL repair by Fanconi-dependent and independent mechanisms.

Fig. 1

(a) Reaction scheme of AAnat-ICL formation. Two acetaldehydes react with deoxyguanine (dG) generating N2-deoxypropanoguanine (PdG), this reacts with a 5’-CpG guanine on the opposite strand. The AAnat-ICL exists in a three-state equilibrium. (b) Replication intermediates generated during ICL repair. (c) Plasmids were replicated in Xenopus egg extract, reaction products were resolved by native agarose gel and visualized by autoradiography. Figure 8 structures (F8), later replication/repair intermediates (RRI), open circle/supercoiled products (OC/SC, grey arrow) are indicated. Six independent experiments. (d) Scheme for the NotI ICL repair assay. Wavy lines: synthesized during repair. (e) Plasmids were replicated in extract, repair intermediates were isolated, NotI-digested, and resolved by denaturing PAGE. Accumulation of the 44 nt product (open arrow) indicates ongoing replication and repair. (*) Product probably generated from end-joining activity in some extracts. Ten independent experiments. (f) Quantification of repair based on the gels in (e), as described in methods (Supplementary Information Methods). Ten independent experiments. (g) Quantification of repair in mock or FANCD2-depleted extract. Based on gel in Extended Data Fig. 2e. Three independent experiments. (h) Scheme of the reduction of ICL-AAnat to ICL-AARED. (i) Plasmids were replicated in extract and products were resolved by native agarose gel. Three independent experiments. (j) Quantification of repair with or without p97i. Based on gel in Extended Data Fig. 3b. Three independent experiments.