Figure 5. Unwinding of flap DNA and three junction substrates by CeWRN-1.

The 5′ flap substrate (A), the 3′ flap (B), and the three-way junction (C) were incubated with varying concentrations of CeWRN-1 at 37 °C for 15 min. Reaction products were analyzed as described in Materials and Methods and were subjected to phosphorimaging analysis. Lane 1 contained no enzyme. Lane 4 contained heat-denatured DNA substrate. Lanes 2 and 3 show a protein titration (5 and 10 nM, respectively). The positions of the initial substrates (IS) and products (P1 or P2) are shown at the left or right. (D–F) Quantification of the data from panels A (D), B (E), and C (F). The percentages of IS and products were calculated using ImageQuant: (white bars) initial substrate, (black bars) single-stranded product (P1), and (gray bars) partial duplex product (P2).