Fig. 7: Interaction between Rad52 and RPA facilitates Rad51 nucleofilament formation by regulating RPA ssDNA binding mode and RPA spacing.

a Rad52-M domain affects the binding dynamics of RPA-WT. (i) schematic of Rad52-M domain, which was 205-345 of Rad52. Representative kymographs of 10-fold RPA/Rad52-M complex at 150 mM NaCl were shown in (ii), and its length analysis (blue) was in (iii), where length dynamic curve with 10-fold RPA-WT (green) from Fig. 3a and 10-fold RPA-ΔWH (red) from Fig. 6b at 150 mM NaCl were added for comparison. RPA/Rad52-M complex was prepared by 30-min pre-incubation. b Boxplot of length (i) or intensity (ii) change proportion within 30–40-min of ssDNA-RPA complexes from 10-fold RPA-WT, and RPA-ΔWH with (red) and without Rad52 M domain (green). Exact p-value (from left to right): 0.0128, 5. 3e-8, 0.2332, 0.7743. Boxplot style: middle line (mean), box range (0.25–0.75), whisker range (min-max), with outliers removed. Statistics: one-way ANOVA (analysis of variation). P-value style: GP: ≥0.05 (ns), <0.05 (*), <0.01 (**), <0.001 (***), <0.0001 (****). c RPA/Rad52-M complex facilitates Rad51 nucleofilament formation on ssDNA–RPA complex. Representative snapshots of ssDNA-RPA complex at 0, 2.5, 5, 7.5, 10 min after Rad51 injection were picked from double-tethered ssDNA Curtains results for 10-Fold RPA with Rad52 M domain (i). Quantification of GFP intensity decrease (ii) during 10-min Rad51 incubation for 10-Fold RPA with Rad52 M domain (red) in RPA-free working buffer and comparison with RPA-WT (green). Data are presented as mean ± SEM in a, c. N represents the total trace number of ssDNA-RPA complex end tracking examined over three times DNA Curtains experiments for each experimental condition in a–c. Source data are provided as a Source Data file.