Promotion of DNA end resection by BRCA1-BARD1 in homologous recombination

• 成果类型: Article
• 署名作者: Salunkhe, Sameer; Daley, James M.; Kaur, Hardeep; Tomimatsu, Nozomi; Xue, Chaoyou; Raina, Vivek B.; Jasper, Angela M.; Rogers, Cody M.; Li, Wenjing; Zhou, Shuo; Mojidra, Rahul; Kwon, Youngho; Fang, Qingming; Ji, Jae-Hoon; Shabestari, Aida Badamchi; Fitzgerald, O'Taveon; Dinh, Hoang; Mukherjee, Bipasha; Habib, Amyn A.; Hromas, Robert; Mazin, Alexander V.; Wasmuth, Elizabeth V.; Olsen, Shaun K.; Libich, David S.; Zhou, Daohong; Zhao, Weixing; Greene, Eric C.; Burma, Sandeep; Sung, Patrick
• 署名单位: University of Texas System; University of Texas at San Antonio; University of Texas System; University of Texas at San Antonio; University of Texas System; University of Texas at San Antonio; NewYork-Presbyterian Hospital; Columbia University; Chinese Academy of Sciences; Tianjin Institute of Industrial Biotechnology, CAS; University of Texas System; University of Texas Southwestern Medical Center; University of Texas System; University of Texas at San Antonio; University of Michigan System; University of Michigan
• 刊物名称: Nature
• ISSN/ISSBN: 0028-6577
• DOI: 10.1038/s41586-024-07910-2
• 发表日期: 2024-10-10
• 页码: 482-+
• 关键词: mechanism; repair; helicases; complex; protein; responses; binding; roles; brca1; cells

摘要

The licensing step of DNA double-strand break repair by homologous recombination entails resection of DNA ends to generate a single-stranded DNA template for assembly of the repair machinery consisting of the RAD51 recombinase and ancillary factors(1). DNA end resection is mechanistically intricate and reliant on the tumour suppressor complex BRCA1-BARD1 (ref. (2)). Specifically, three distinct nuclease entities-the 5-3 ' exonuclease EXO1 and heterodimeric complexes of the DNA endonuclease DNA2, with either the BLM or WRN helicase-act in synergy to execute the end resection process(3). A major question concerns whether BRCA1-BARD1 directly regulates end resection. Here, using highly purified protein factors, we provide evidence that BRCA1-BARD1 physically interacts with EXO1, BLM and WRN. Importantly, with reconstituted biochemical systems and a single-molecule analytical tool, we show that BRCA1-BARD1 upregulates the activity of all three resection pathways. We also demonstrate that BRCA1 and BARD1 harbour stand-alone modules that contribute to the overall functionality of BRCA1-BARD1. Moreover, analysis of a BARD1 mutant impaired in DNA binding shows the importance of this BARD1 attribute in end resection, both in vitro and in cells. Thus, BRCA1-BARD1 enhances the efficiency of all three long-range DNA end resection pathways during homologous recombination in human cells.