Figure 7.
RPA Depletion Stimulates Pol α-Dependent Leading-Strand Restart
(A) Reaction scheme for bead-bound replication assays.
(B–D) Reactions performed as illustrated in (A) on undamaged and 3 kb CPDLEAD templates (20 nM Pol α throughout). Products were separated through denaturing gels (B), and lane profiles for uncut (C) and immobilized AvrII-treated samples (D) are shown.
(E and F) Reaction performed as illustrated in (A) but with 5 nM Pol α in step 1 and varying concentrations of Pol α in step 2 as indicated. AvrII digested products were separated though denaturing gels (E), and lane profiles for the immobilized samples (F) are shown.
(G) Model of the initial response of the replisome to (i) lagging- and (ii) leading-strand CPDs.