Figure 3.
Response of the Replisome to a Leading-Strand CPD
(A) Schematic showing the 3 kb CPDLEAD template and the predicted replication products of leading-strand lesion bypass by re-priming. In this and all subsequent figures the putative restart product is shown as a dashed red line.
(B and C) Comparison of replication products from undamaged and 3 kb CPDLEAD templates in the presence (B) and absence of Pol δ (C). (C) In this and all subsequent experiments entirely lacking Pol δ, the reaction buffer contained 117 mM potassium glutamate.
(D) Two-dimensional gel of a replication reaction performed on the 3 kb CPDLEAD template (top), together with a schematic of the products generated (bottom).
(E and F) Pulse-chase experiments on undamaged (E) and 4.5 kb CPDLEAD templates (F). The chase was added 14 min 50 s after replication was initiated.
(G and H) Quantitation of pulse-chase experiments on undamaged (G) and 4.5 kb CPDLEAD templates (H) performed as in (E) and (F). “Resolved” is the sum of full-length and uncoupled products. Error bars represent the SEM from four experiments.