Figure 6.

Holliday junction resolution can occur during branch migration. Junction X1 (5′-³²P-end labeled in strand 2) was incubated with RuvC (10 nm), or with RuvA (20 nm), RuvB (600 nm) and RuvC (10 nm) for 30 min at 37°C, as indicated. In this experiment, resolution was permitted by inclusion of Mg²⁺, whereas branch migration was either blocked using AMP–PNP (lanes b and c) or slowed by the presence of a mixture of ATPγS and ATP (lane d). (Lane e) Reaction carried out in the absence of nucleoside triphosphates. ³²P-Labeled products were analyzed by denaturing PAGE, alongside A + G and C > T sequence ladders, obtained by chemical cleavage of X1 strand 2. The thymine at the base of the junction is indicated by 0, and +1 and +6 indicate cleavage sites in the short arm away from the crossover.