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. 2019 Dec 4;48(2):694–708. doi: 10.1093/nar/gkz1052

Figure 4.

Figure 4.

RAD52 annealing and homology search are stimulated by DSS1. (A) Gel-based SSA assay using complementary fluorescent 3′-OH substrates in the presence of indicated concentration of RAD52 or RAD52–DSS1 complex. The position of substrate and product of annealing reaction are indicated. (B) Quantification of the annealing reactions from three independent experiments. The mean values ± SD were plotted. (C) FRET based ssDNA annealing experiments of complementary 28-mer oligonucleotides labelled with Cy3 and Cy5 dyes, respectively in the presence of RAD52 (8 nM, blue) and RAD52-DSS1 complex (8 nM-40 nM, orange). (D) FRET based ssDNA annealing experiment of complementary 28-mer oligonucleotides labelled with Cy3 and Cy5 dyes in the presence of RPA (2 nM), respectively in the presence of stoichiometric amounts of RAD52-RPA (8–2 nM, blue) and RAD52-RPA-DSS1 (8–2–40 nM, orange) complex. These data are fit to a double exponential (solid lines). (E) Scheme of in vitro D-loop reaction and representative agarose gel showing the formation of D-loop using indicated amounts of RAD52 or RAD52–DSS1. (F) Quantification of the D-loop reaction from three independent experiments. The mean values ± SD from were plotted.