Figure 1.

The Replisome Rapidly and Efficiently Bypasses a Lagging-Strand CPD

(A) Schematic of AhdI- and BamHI-linearized undamaged templates and the predicted replication products. In this and all subsequent figures; red: leading-strands; blue: lagging-strands; the position of the ARS306 origin of replication is marked, Ori. The location of the SmaI restriction site is indicated.

(B) Standard replication reactions performed on the templates illustrated in (A). Unless stated otherwise, this and all subsequent standard replication assays contained 217 mM potassium glutamate. Templates were prepared by linearizing maxiprep DNA (Maxi) or undamaged plasmids prepared using the same method used to generate CPD containing plasmids (Ligated).

(C) Schematic of the 6.7 kb CPD^LAG template and the predicted replication products of lagging-strand lesion bypass.

(D) Replication reaction comparing undamaged and 6.7 kb CPD^LAG templates.

(E) Schematic of the 5.1 kb CPD^LAG template.

(F) Pulse-chase experiment on undamaged and 5.1 kb CPD^LAG templates. The chase was added at 2 min 50 s.

(G) Quantitation of pulse-chase experiments performed as in (F). Error bars represent the SEM from four experiments. Data were fit to a linear regression. Dashed line indicates the distance from Ori to CPD^LAG (5.1 kb).