Role of BRCA2 DNA-binding and C-terminal domain in its mobility and conformation in DNA repair

eLife

Yongxin Liang¹  Alex N Zelensky¹  Hanny Odijk²  Arshdeep Sidhu²  Claire Wyman²  Maarten W Paul²  Roland Kanaar²  Sarah E van Rossum-Fikkert² 

[1] Department of Radiation Oncology, Erasmus University Medical Center, Rotterdam, Netherlands;Department of Molecular Genetics, Oncode Institute, Erasmus MC Cancer Institute, Erasmus University Medical Center, Rotterdam, Netherlands;

关键词: DNA repair;    homologous recombination;    BRCA2;    scanning force microscopy;    single-molecule microscopy;    nuclear foci;   

DOI  :  10.7554/eLife.67926

来源: DOAJ

【 摘 要 】

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.

【 授权许可】

Unknown