Extended Data Fig. 8: X3CDX2 complex promotes RAD51 filament assembly.
From: Structural insight into how RAD51 paralog exchange regulates RAD51 filament formation
a, ssDNA binding of BC, X3C and DX2 complexes and co-complexes of DX2 with BC (BCDX2) and X3C (X3CDX2). Results from 6 independent experiments were plotted as mean values ± s.d. b, ssDNA binding of WT X3CDX2 or X3CDX2 carrying RAD51D_R266A mutants at the indicated protein concentrations in the presence of AMP-PNP. Results from three independent experiments were plotted as mean values ± s.d. c, ssDNA binding of RAD51-X3C (20 nM) in the presence of increasing concentrations of DX2 complex and AMP-PNP. Results from three independent experiments were plotted as mean values ± s.d. d, Representative micrographs depicting RAD51 nucleoprotein filaments seen by negative stain electron microscopy in absence or presence of indicated concentrations of X3CDX2 complex. Scale is in nanometers. Data represent 49 electron micrographs for each sample captured in two independent experiments. e, Strand exchange in the presence of 30 nM RPA with 100 nM RAD51-X3C alone or in the presence of increasing concentrations of DX2 complex. Reactions were performed in the presence of AMP-PNP. Results from three independent experiments were plotted as mean values ± s.d. f, ssDNA binding of X3CDX2 or X3C complexes carrying either WT or indicated XRCC3 mutants in the presence of AMP-PNP. Results from three independent experiments were plotted as mean values ± s.d. g, ssDNA binding of RAD51 (20 nM) alone or and indicated conc. of X3CDX2 complexes carrying either WT or XRCC3 mutants in the presence of AMP-PNP. Results from three independent experiments were plotted as mean values ±. s.d. h, Strand exchange in the presence of 30 nM RPA with 100 nM RAD51 alone or in the presence of equimolar X3CDX2 complexes carrying either WT or XRCC3 mutants. Reactions were performed in presence of AMP-PNP. Results from four independent experiments were plotted as mean values ± s.d.
