2019
DOI: 10.1080/15384101.2019.1691802
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Determination of the number of RAD51 molecules in different human cell lines

Abstract: Knowledge about precise numbers of specific molecules is necessary for understanding and verification of biological pathways. The RAD51 protein is central in the repair of DNA doublestrand breaks (DSBs) by homologous recombination repair and understanding its role in cellular pathways is crucial to design mechanistic DNA repair models. Here, we determined the number of RAD51 molecules in several human cell lines including primary fibroblasts. We showed that between 20000 to 100000 of RAD51 molecules are availa… Show more

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Cited by 7 publications
(6 citation statements)
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“…In CMM patients' cells, the WT-RAD51 protein is reduced (42-67% of WT-RAD51 levels in the intrafamilial controls) without any phenotypic evidence for alterations in its homologous recombination activity: in our CMM cohort, none of the patients with RAD51 pathogenic variants presented fertility problems, Fanconi anemia, or cancer. In line with this observation, the cellular levels of RAD51 are thought to exceed the cell's needs for homologous recombination (35). Our findings strongly support that both truncating and non-truncating RAD51-CMM variants result in a loss of function selectively associated with the CMM phenotype.…”
Section: Discussionsupporting
confidence: 80%
“…In CMM patients' cells, the WT-RAD51 protein is reduced (42-67% of WT-RAD51 levels in the intrafamilial controls) without any phenotypic evidence for alterations in its homologous recombination activity: in our CMM cohort, none of the patients with RAD51 pathogenic variants presented fertility problems, Fanconi anemia, or cancer. In line with this observation, the cellular levels of RAD51 are thought to exceed the cell's needs for homologous recombination (35). Our findings strongly support that both truncating and non-truncating RAD51-CMM variants result in a loss of function selectively associated with the CMM phenotype.…”
Section: Discussionsupporting
confidence: 80%
“…HR of DNA repair in eukaryotic cells is mainly conducted by two recombinases, Rad51 and Dmc1. Rad51 is not only responsible for DNA double-strand break repair in mitosis, for example, repairing interrupted replication forks occurring in G2/S phase ( 63 , 64 ) but has also been identified during meiosis to facilitate HR mediated by Dmc1 ( 18 , 65 ). However, the only function of Dmc1 is to mediate HR between parental chromosome for genetic diversity in meiosis I ( 66 ).…”
Section: Discussionmentioning
confidence: 99%
“…We also attempted to follow by live imaging the recruitment of another repair factor, RAD51, involved in DSB repair by homologous recombination (HR). However, even when transfecting cells with a plasmid encoding GFP-RAD51 under the control of a weak promoter ( Foertsch et al, 2019 ), the levels of expression were still too high and led to the formation of RAD51 filaments even in undamaged cells, which hampered the study of RAD51 relocalization after laser micro-irradiation. Instead, we assessed the recruitment of two endogenous HR repair factors, phosphorylated RPA (pRPA32 Ser4/8) and RAD51, by immunofluorescence in RPE-1 cells stably expressing macroH2A1.2-GFP ( Figure 2H ).…”
Section: Anticipated Resultsmentioning
confidence: 99%