Differential Requirements for the RAD51 Paralogs in Genome Repair and Maintenance in Human Cells

English version

Autoři: Edwige B. Garcin ^aff001; Stéphanie Gon ^aff001; Meghan R. Sullivan ^aff002; Gregory J. Brunette ^aff002; Anne De Cian ^aff003; Jean-Paul Concordet ^aff003; Carine Giovannangeli ^aff003; Wilhelm G. Dirks ^aff004; Sonja Eberth ^aff004; Kara A. Bernstein ^aff002; Rohit Prakash ^aff005; Maria Jasin ^aff005; Mauro Modesti ^aff001
Působiště autorů: Cancer Research Center of Marseille; CNRS; Inserm; Institut Paoli-Calmettes; Aix-Marseille Université, Marseille, France ^aff001; Department of Microbiology and Molecular Genetics, University of Pittsburgh School of Medicine and UPMC Hillman Cancer Center, Pittsburgh, Pennsylvania, United States of America ^aff002; Museum National d'Histoire Naturelle, Inserm U1154, CNRS UMR 7196, Sorbonne Universités, Paris, France ^aff003; Department of Human and Animal Cell Lines, Leibniz-Institute DSMZ-German, Collection of Microorganisms and Cell Cultures, Braunschweig, Germany ^aff004; Developmental Biology Program, Memorial Sloan Kettering Cancer Center, New York, New York, United States of America ^aff005
Vyšlo v časopise: Differential Requirements for the RAD51 Paralogs in Genome Repair and Maintenance in Human Cells. PLoS Genet 15(10): e32767. doi:10.1371/journal.pgen.1008355
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.pgen.1008355

Souhrn

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. 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.

Klíčová slova:

Genetic loci – Polymerase chain reaction – Analysis of variance – Plasmid construction – Complement system – Guide RNA – Cell disruption

Zdroje

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