Figure 4.
Polκ-PCNA interaction and its CDs are important for insertion during FA-mediated aldehyde ICL repair. (A) Schematic representation of xlPolκ indicating its functional domains. Mutations are indicated and described in the table (bottom). (B) Western blot analysis of mock, Polκ-depleted (ΔPolκ), and Polκ-depleted egg extract supplemented with WT Polκ or mutants, alongside a titration of undepleted extract. (C) Repair intermediates of pICL-ACRRED replicated in mock, ΔPolκ, and ΔPolκ egg extract supplemented with WT or indicated mutant Polκ were extracted, digested with AflIII, and separated on a 7% urea–PAGE gel. Grey arrows: −1 stalling products. Quantifications of lesion bypass assays on pICL-AARED (D), pICL-ACRRED (E), and pICL-Pt (F) repair reactions in mock, ΔPolκ, and ΔPolκ + WT or mutant Polκ extract. Based on gels in Supplementary Figs S7A and B, and S8A.