TY - JOUR TI - Role of BRCA2 DNA-binding and C-terminal domain in its mobility and conformation in DNA repair AU - Paul, Maarten W AU - Sidhu, Arshdeep AU - Liang, Yongxin AU - van Rossum-Fikkert, Sarah E AU - Odijk, Hanny AU - Zelensky, Alex N AU - Kanaar, Roland AU - Wyman, Claire A2 - Spies, Maria A2 - Tyler, Jessica K A2 - Esashi, Fumiko A2 - Hengel, Sarah R VL - 10 PY - 2021 DA - 2021/07/13 SP - e67926 C1 - eLife 2021;10:e67926 DO - 10.7554/eLife.67926 UR - https://doi.org/10.7554/eLife.67926 AB - Breast cancer type two susceptibility protein (BRCA2) is an essential protein in genome maintenance, homologous recombination (HR), and replication fork protection. Its function includes multiple interaction partners and requires timely localization to relevant sites in the nucleus. We investigated the importance of the highly conserved DNA-binding domain (DBD) and C-terminal domain (CTD) of BRCA2. We generated BRCA2 variants missing one or both domains in mouse embryonic stem (ES) cells and defined their contribution in HR function and dynamic localization in the nucleus, by single-particle tracking of BRCA2 mobility. Changes in molecular architecture of BRCA2 induced by binding partners of purified BRCA2 were determined by scanning force microscopy. BRCA2 mobility and DNA-damage-induced increase in the immobile fraction were largely unaffected by C-terminal deletions. The purified proteins missing CTD and/or DBD were defective in architectural changes correlating with reduced HR function in cells. These results emphasize BRCA2 activity at sites of damage beyond promoting RAD51 delivery. KW - DNA repair KW - homologous recombination KW - BRCA2 KW - scanning force microscopy KW - single-molecule microscopy KW - nuclear foci JF - eLife SN - 2050-084X PB - eLife Sciences Publications, Ltd ER -