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. 2001 Dec 15;15(24):3308–3318. doi: 10.1101/gad.935501

Figure 4.

Figure 4

Mediator function of Rad51B–Rad51C. (A) Schematic of the homologous DNA pairing and strand exchange reaction using φX174 DNA substrates. Linear duplex is paired with the homologous ssDNA circle to yield a joint molecule. DNA strand exchange, if successful over the length (5.4 kb) of the DNA molecules, results in the formation of the nicked circular duplex. (B) Rad51-mediated DNA pairing and strand exchange was carried out with RPA (panel I) or without it (panel II). In panel I, the ssDNA was preincubated with Rad51 (R51) before RPA was added. The concentrations of the reaction components were: Rad51, 7.5 μM; RPA, 1.5 μM; ssDNA, 30 μM nucleotides; linear duplex, 15 μM base pairs. (C) In the DNA strand exchange reaction in panel I, the ssDNA was incubated with both Rad51 (R51) and RPA simultaneously, and in the reaction in panel II, the ssDNA was incubated with Rad51, RPA and Rad51B–Rad51C (B–C) simultaneously. The concentration of Rad51B–Rad51C was 0.8 μM, while the concentrations of the other components were exactly as those in B. In panel III, the amounts of nicked circular duplex in the reactions represented in B panel I (filled squares) and panel II (open circles) and in C panel I (filled circles) and panel II (open squares) are plotted. In panel IV, the amounts of total reaction products (sum of joint molecules and nicked circular duplex) in the reactions represented in B panel I (filled squares) and panel II (open circles) and in C panel I (filled circles) and panel II (open squares) are plotted.