Fig. 1: BRCA1–BARD1 directly promotes resection by WRN-DNA2-RPA.
From: Mechanism of BRCA1–BARD1 function in DNA end resection and DNA protection
a, Recombinant BRCA1 (top) and the BRCA1–BARD1 (bottom) complex used in this study. The polyacrylamide gels were stained with Coomassie Brilliant Blue. b, Resection assays with WRN-DNA2-RPA, and its stimulation by BRCA1–BARD1. Top right, a schematic of the assay. Red asterisks (*) represent the position of the radioactive labels. Top, quantitation of DNA degradation. Averages shown; error bars, s.e.m. (lanes 4–7); n = 2 for the reaction containing 60 nM of BRCA1–BARD1 and n = 3 for all the other samples. Bottom, representative assays. c, Resection assays with WRN-DNA2-RPA, in the absence or presence of either BRCA1–BARD1 or BRCA1. Top right, a schematic of the assay. Red asterisks (*) represent the position of the radioactive labels. Top, quantitation of DNA degradation. Averages shown; error bars, s.e.m.; n = 3. **P = 0.0055, two-tailed t-test. Bottom, representative assays. d, Resection assays with DNA2-RPA, and either wild-type (WT) WRN or exonuclease-dead WRN E84A, in the absence or presence of BRCA1–BARD1. Top, quantitation of the substrate utilization. Averages shown; error bars, s.e.m.; n = 3. Bottom, representative assays. Source data are provided in Supplementary Fig. 1. NS, not significant.
