Fig. 4.

Degradation and idling by WT and mutant Pols on primer templates containing internally incorporated GCV. (A and B) Radiolabeled primer template T3, which contains GCV plus dC (A), or T4, whose major species contains GCV plus dC plus dA (B), was incubated with WT or the indicated mutant Pol in the absence of TPs and the presence of UL44 for various times, and the products were analyzed by polyacrylamide gel electrophoresis and autoradiography. The enzymes used are indicated at the top of each panel, and the times incubated in minutes at the top of each lane. Lanes in B marked T3 contain untreated T3 as a control and size marker. (C) Unlabeled primer template T3 was incubated with dATP plus [α-³²P]-dATP using each of the Pols indicated at the top of the panel in the presence of UL44 at 37 °C for 10 min, and the products were analyzed by TLC and autoradiography. Co., T3 incubated with dATP plus [α-³²P]-dATP without any protein. The positions of dATP and dAMP were determined by visualizing unlabeled standards under UV light and are indicated by arrows to the right of the panel.