Fig. 4.

Hop2-Mnd1 allows Dmc1 to exchange strands with homology embedded within nonhomologous DNA. (A) Schematic of the D-loop assay employing a linear ssDNA and either lds (Left) or nc (Right). (B) Gel images of the D-loop assay with naked lss. (C) Gel images of the D-loop assay with RPA-precoated lss. In B and C, lds (Left) or nc (Right) was used. In B, lss was preincubated with Dmc1, then mixed with auxiliary factor(s) and dsDNA. In C, lss was precoated with RPA, then mixed with Dmc1, auxiliary factor(s), and dsDNA. After the addition of dsDNA, the reaction was incubated at 30 °C for 1 h. Yield is expressed as the percentage of the signal for the products to the sum of the products and either lds or nc. Dmc1, 5 µM; Hop2-Mnd1, 0.25 µM; lds, 5 µM; lss, 5 µM; nc, 5 µM; RPA, 1 µM; Swi5-Sfr1, 0.5 µM in B and 4 µM in C. For the graphs in B and C, mean values ± SD from three independent experiments are shown.