The complexity of double-strand break ends is a factor in the repair pathway choice.
The repair of double-strand breaks in mammalian cells is carried out by two pathways: homologous recombination and nonhomologous end joining. The factors that regulate the mechanism through which a specific repair pathway is activated are still not clearly defined. To study whether the complexity of the double-strand break ends is a factor that determines the choice of the repair pathway, we examined the involvement of homologous recombination by the formation of Rad51 foci in human HeLa cells treated with bleomycin and ionizing radiation. The quantity of double-strand breaks was determined by gel electrophoresis and the formation of gamma-H2AX foci. Two hours after treatment with low doses of the agents that induced similar quantities of double-strand breaks that could be repaired effectively by the cells, Rad51 foci were observed only in the irradiated cells. Rad51 foci appeared in bleomycin-treated cells after prolonged exposure to the drug when the cells were arrested in the G2 phase of the cell cycle. Since bleomycin produces double-strand breaks that are less complex than the breaks induced by ionizing radiation, these results indicate that the complexity of the break ends is a factor in the choice of repair pathway and that homologous recombination is recruited in the repair of breaks with more complex multiply damaged ends during the late S and G2 phases of the cell cycle.
Mladenov
Emil
E
Institute of Molecular Biology, Bulgarian Academy of Sciences, Sofia 1113, Bulgaria.
Kalev
Peter
P
Anachkova
Boyka
B
D001761:Bleomycin / Q000494:pharmacology; D002453:Cell Cycle; D002875:Chromosomes / Q000648:ultrastructure; D053903:DNA Breaks, Double-Stranded; D004260:DNA Repair; D004307:Dose-Response Relationship, Radiation; D005434:Flow Cytometry; D016195:G2 Phase; D006367:HeLa Cells; D006657:Histones / Q000378:metabolism; D006801:Humans; D008856:Microscopy, Fluorescence / Q000379:methods; D051135:Rad51 Recombinase / Q000378:metabolism; D011995:Recombination, Genetic; D013997:Time Factors
C468783:H2AX protein, human; D006657:Histones; D001761:Bleomycin; C495153:RAD51 protein, human; D051135:Rad51 Recombinase