Fig. 3: A pairing-initiated abortive DNA strand exchange assay indicates that C1 intermediates accumulate in the L2 loop mutant.
From: Real-time tracking reveals catalytic roles for the two DNA binding sites of Rad51
a Schematic diagram of a pairing-initiated abortive DNA strand exchange assay. EDTA was added to the ongoing pairing reactions to chelate Mg2+, leading to dissociation of Rad51 from the intermediates. Fluorescence emission increases if the C1 intermediate is converted to substrates, but does not change substantially if the C2 intermediates are converted to products. b–e Time courses of the pairing-initiated abortive assay: b wild-type Rad51; c wild-type Rad51 with Swi5-Sfr1; d Rad51-L2; and e Rad51-L2 with Swi5-Sfr1. EDTA was added in the reaction mixture 5 (blue), 10 (red), or 20 (green) min, as indicated by arrows, after the strand pairing reaction started. f Time course of accumulation (% of the input) of the C1 intermediate. wild-type Rad51; blue, wild-type Rad51 with Swi5-Sfr1; red, Rad51-L2; green and Rad51-L2 with Swi5-Sfr1; purple. Data are expressed as the mean ± s.d. (n = 3 independent experiments). Source data are provided as a Source data file.
