Induction of base damages representing a high risk site for double-strand DNA break formation in genomic DNA by exposure of cells to DNA damaging agents.
DNA repair is known as a defense mechanism against genotoxic insults. However, the most lethal type of DNA damages, double-strand DNA breaks (DSBs), can be produced by DNA repair. We have previously demonstrated that when long patch base excision repair attempts to repair a synthetic substrate containing two uracils, the repair produces DSBs (Vispe, S. and Satoh, M. S. (2000) J. Biol. Chem. 275, 27386-27392 and Vispe, S., Ho, E. L., Yung, T. M., and Satoh, M. S. (2003) J. Biol. Chem. 278, 35279-35285). In this synthetic substrate, the two uracils are located on the opposite DNA strands (separated by an intervening sequence stable at 37 degrees C) and represent a high risk site for DSB formation. It is not clear, however, whether similar high risk sites are also induced in genomic DNA by exposure to DNA damaging agents. Thus, to investigate the mechanisms of DSB formation, we have modified the DSB formation assay developed previously and demonstrated that high risk sites for DSB formation are indeed generated in genomic DNA by exposure of cells to alkylating agents. In fact, genomic DNA containing alkylated base damages, which could represent high risk sites, are converted into DSBs by enzymes present in extracts prepared from cells derived from clinically normal individuals. Furthermore, DSBs are also produced by extracts from cells derived from ataxia-telangiectasia patients who show cancer proneness due to an impaired response to DSBs. These results suggest the presence of a novel link between base damage formation and DSBs and between long patch base excision repair and human diseases that occur due to an impaired response to DSB.
Ho
Erick L Y
EL
Division of Health and Environmental Research, Laval University Medical Centre (CHUL) and Department of Anatomy and Physiology, Faculty of Medicine, Laval University, Quebec, Quebec, Canada.
Parent
Marianne
M
Satoh
Masahiko S
MS
D000477:Alkylating Agents / Q000633:toxicity; D001483:Base Sequence; D002460:Cell Line; D004247:DNA / Q000187:drug effects / Q000235:genetics* / Q000528:radiation effects; D004249:DNA Damage / Q000187:drug effects* / Q000528:radiation effects; D004270:DNA, Circular / Q000187:drug effects / Q000235:genetics; D005720:Gamma Rays; D006367:HeLa Cells; D006801:Humans; D008769:Methylnitronitrosoguanidine / Q000633:toxicity*; D016133:Polymerase Chain Reaction; D014498:Uracil
D000477:Alkylating Agents; D004270:DNA, Circular; D008769:Methylnitronitrosoguanidine; D014498:Uracil; D004247:DNA