Fig. 10: Rad51-ED is severely affected in binding to damaged DNA and Sister Chromatid Junction formation.

A rad51-KN and rad51-ED are defective in the accumulation of Sister Chromatid Junctions (SCJ), as determined by 2D gel analysis of X-shaped molecules in sgs1∆ (W303sgs1), sgs1∆ rad51-KN (W303sr305) and sgs1∆ rad51-ED (W303sr135) cells synchronized in G1 and released in the presence of 0.033% MMS. A schematic representation of the migration pattern of X-shaped (SCJs) and Y-shaped (replication intermediates) molecules, as well as the DNA content analysis of the different cultures along the time course, is shown on the left. The panel on the right shows the amounts of X-shaped molecules relative to the total amount of molecules at the indicated times. The highest value is set at 100, and the mean and SEM from four independent time courses are given. The area under the curve (AUC) is also plotted. Two and three asterisks indicate statistically significant differences according to a One-Anova (Bonferroni) test (P < 0.001 and 0.0001, respectively). B Rad51 binding to replicative DNA lesions as determined by ChEC analysis of exponentially growing RAD51-MN (wt) (wR51MN-2), rad51-ED-MN (wR51-135MN) and rad51-KN-MN (wR51-305MN) cells incubated with and without 0.05% MMS for 2 h. Total DNA from cells permeabilized and treated or not with Ca²⁺ for 15 min is shown, as well as the DNA digestion profiles. ChEC experiments were repeated three times with similar results. Source data are provided as a Source Data file.