Abstract
Deficiency in several of the classical human RAD51 paralogs [RAD51B, RAD51C, RAD51D, XRCC2 and XRCC3] is associated with cancer predisposition and Fanconi anemia. To investigate their functions, isogenic disruption mutants for each were generated in non-transformed MCF10A mammary epithelial cells and in transformed U2OS and HEK293 cells. In U2OS and HEK293 cells, viable ablated clones were readily isolated for each RAD51 paralog; in contrast, with the exception of RAD51B, RAD51 paralogs are cell-essential in MCF10A cells. Underlining their importance for genomic stability, mutant cell lines display variable growth defects, impaired sister chromatid recombination, reduced levels of stable RAD51 nuclear foci, and hyper-sensitivity to mitomycin C and olaparib, with the weakest phenotypes observed in RAD51B-deficient cells. Altogether these observations underscore the contributions of RAD51 paralogs in diverse DNA repair processes, and demonstrate essential differences in different cell types. Finally, this study will provide useful reagents to analyze patient-derived mutations and to investigate mechanisms of chemotherapeutic resistance deployed by cancers.
Highlights
RecA family recombinases are universally found in the three domains of life: RadA in Archaea, RecA in Bacteria and RAD51/DMC1 in Eukarya [1]
Two RAD51 paralogs have been clearly identified as tumor suppressors and the other three paralogs have been found to be mutated in some tumors
RAD51C, RAD51D, XRCC2 and XRCC3 are essential for survival in MCF10A but RAD51B is dispensable
Summary
RecA family recombinases are universally found in the three domains of life: RadA in Archaea, RecA in Bacteria and RAD51/DMC1 in Eukarya [1]. Additional RecA-like genes have evolved in the three domains of life presumably after duplication and divergent evolution of a common ancestor [1,4] While structurally related, these paralogs do not promote homology recognition and strand exchange but rather act in part as accessory factors to the core recombinases [5,6,7,8,9,10,11,12,13]. SWSAP1, a non-classical RAD51 paralog, was recently identified as a component of the Shu complex [26,27,28]
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
