Extended Data Fig. 4: Restriction of RAD51-dsDNA association by X3C.
From: Structural insight into how RAD51 paralog exchange regulates RAD51 filament formation
a, 80 nt ssDNA and 80 bp dsDNA binding of X3C complex at indicated concentrations in the presence of AMP-PNP was assessed by electrophoretic mobility shift assays. Results from three independent experiments were plotted as mean values ± s.d. b, 80 nt ssDNA binding and c, 80 bp dsDNA binding of RAD51 and RAD51-X3C complex at indicated RAD51 concentrations in presence of ATP assessed by electrophoretic mobility shift assays. Results from three independent experiments were plotted as mean values ± s.d. d, Averaged and fitted (v=Vmax*[S]/{Kd + [S]}) line plots for (i) 80 nt ssDNA binding, (ii) 80 bp dsDNA and (iii) 60 bp RNA:DNA hybrids binding of RAD51 and RAD51-X3C complex at indicated RAD51 concentrations in the presence of AMP-PNP assessed by electrophoretic mobility shift assays. Results from three independent experiments were plotted as mean values ± s.d. e, ssDNA binding and f, dsDNA binding of RAD51 in the presence of increasing concentrations of X3C and 1 mM ATP. Results from three independent experiments were plotted as mean values ± s.d. g, ssDNA binding of RAD51 in the presence of increasing concentrations of X3C and 1 mM AMP-PNP following preincubation with excess circular dsDNA. Results from three independent experiments were plotted as mean values ± s.d. h, Reaction schematic and strand exchange with increasing concentrations of either RAD51 alone or X3C-RAD51 in the presence of AMP-PNP, results from four independent experiments were plotted as mean values ± s.e., i, Strand exchange with RAD51 alone or in the presence of X3C complexes carrying either WT or XRCC3 mutants. Reactions were performed in presence of AMP-PNP and results from four independent experiments were plotted as mean values ± s.e.
