A tumour-derived mutant allele of XRCC2 preferentially suppresses homologous recombination at DNA replication forks.
Homologous recombination repair (HRR) is required for both the repair of DNA double strand breaks (DSBs) and the maintenance of the integrity of DNA replication forks. To determine the effect of a mutant allele of the RAD51 paralog XRCC2 (342delT) found in an HRR-defective tumour cell line, 342delT was introduced into HRR proficient cells containing a recombination reporter substrate. In one set of transfectants, expression of 342delT conferred sensitivity to thymidine and mitomycin C and suppressed HRR induced at the recombination reporter by thymidine but not by DSBs. In a second set of transfectants, the expression of 342delT was accompanied by a decreased level of the full-length XRCC2. These cells were defective in the induction of HRR by either thymidine or DSBs. Thus 342delT suppresses recombination induced by thymidine in a dominant negative manner while recombination induced by DSBs appears to depend upon the level of XRCC2 as well as the expression of the mutant XRCC2 allele. These results suggest that HRR pathways responding to stalled replication forks or DSBs are genetically distinguishable. They further suggest a critical role for XRCC2 in HRR at replication forks, possibly in the loading of RAD51 onto gapped DNA.
Mohindra
Atul
A
Institute for Cancer Studies, University of Sheffield, School of Medicine, Sheffield, UK.
Bolderson
Emma
E
Stone
Jason
J
Wells
Michael
M
Helleday
Thomas
T
Meuth
Mark
M
D020137:Base Pair Mismatch; D002166:Camptothecin / Q000494:pharmacology; D004247:DNA / Q000235:genetics / Q000378:metabolism; D004260:DNA Repair / Q000235:genetics; D004261:DNA Replication / Q000187:drug effects / Q000235:genetics*; D004268:DNA-Binding Proteins / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism*; D005260:Female; D017930:Genes, Reporter; D006801:Humans; D007890:Leiomyosarcoma / Q000235:genetics; D016685:Mitomycin / Q000494:pharmacology; D009154:Mutation; D011995:Recombination, Genetic; D012509:Sarcoma / Q000235:genetics; D013936:Thymidine / Q000378:metabolism; D014407:Tumor Cells, Cultured; D014594:Uterine Neoplasms / Q000235:genetics
D004268:DNA-Binding Proteins; C113187:XRCC2 protein, human; D016685:Mitomycin; D004247:DNA; D013936:Thymidine; D002166:Camptothecin