Fig. 1: Facilitation of BRCA2-mediated RAD51 Targeting onto ssDNA over dsDNA by DSS1. | Nature Communications

Fig. 1: Facilitation of BRCA2-mediated RAD51 Targeting onto ssDNA over dsDNA by DSS1.

From: DSS1 restrains BRCA2’s engagement with dsDNA for homologous recombination, replication fork protection, and R-loop homeostasis

Fig. 1

(See also Supplementary Fig. 1). a Schematic representation of BRCA2, miBRCA2, and DBD with various functional domains and interaction partners (top). Overall view of the mouse BRCA2-DBD (light green) in complex with DSS1 (orange; PDB: 1MIU) with three potential interfaces (I, II, and III, bottom). b Schematic representation of the magnetic bead-based pulldown assay to investigate RAD51 loading onto biotin-labeled ssDNA (dT83) in the presence of excessive dsDNA. c Western blot analyses to monitor RAD51 loading by miBRCA2 (lanes 4–6), miBRCA2-DSS1 (lanes 7–9), and miBRCA2-SS18A (lanes 10–12) onto the ssDNA substrate at 45 mM KCl condition. Bead without dT83 conjugation (lane 1) and pulldown without dsDNA (lane 2) served as controls. The mean values (±SD) from three independent experiments were correspondingly plotted at the bottom panel. ns not significant; ****p ≤ 0.0001 (two-way ANOVA). d Quantification (mean ± SD) of dsDNA and ssDNA binding by miBRCA2, miBRCA2-DSS1, and miBRCA2-DSS18A from three independent experiments shown in Supplementary Fig. 1k at 90 mM KCl condition. miBRCA2 has significantly higher affinity to ssDNA and dsDNA than either miBRCA2-DSS1 or miBRCA2-DSS18A with ****P ≤ 0.0001 (two-way ANOVA). Source data are provided as a Source Data file.

Back to article page