Fig. 2: DSS1 modulates BRCA2-HDOB1 DNA binding. | Nature Communications

Fig. 2: DSS1 modulates BRCA2-HDOB1 DNA binding.

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

Fig. 2

(See also Supplementary Figs. 2–7). a Schematic representation of DBD-DSS1 with three interfaces and various functional domains used in the study. b Representative gel of dsDNA (5 nM) and ssDNA (5 nM) binding by GST-HDOB1, GST-HD, GST-OB1, and OB23 and GST tagged DSS11–36-HDOB1 at 90 mM KCl condition from three independent experiments. c Schematic depicting HDOB1 with wild-type or various mutants of DSS1 (top), and the complex stability was tested under increasing amounts of salt (KCl) in the GST bead pull-down experiments (bottom). Representative Western blot analyses to monitor His (6)-HDOB1 and GST-DSS1 levels with anti-His or GST antibodies, respectively, from three independent experiments. d Quantification (mean ± SD) of ssDNA (5 nM) binding at 45 mM KCl condition by GST-HDOB1 and GST tagged complexes (HDOB1-DSS1, HDOB1-DSS11–36, HDOB1-DSS11–45, HDOB1-DSS11–54, and HDOB1-DSS1R57Q) from three independent experiments shown in Supplementary Fig. 4a, ****p ≤ 0.0001 (two-way ANOVA). e Quantification (mean ± SD) of dsDNA (5 nM) binding at 45 mM KCl condition by GST-HDOB1 and GST tagged complexes (HDOB1-DSS1, HDOB1-DSS11–36, HDOB1-DSS11–45, HDOB1-DSS11–54, and HDOB1-DSS1R57Q) from three independent experiments shown in Supplementary Fig. 4b, *p ≤ 0.05; ****p ≤ 0.0001 (two-way ANOVA). Source data are provided as a Source Data file.

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