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• W2059920795 abstract "Willers, H., Husson, J., Lee, L. W., Hubbe, P., Gazemeirer, F., Powell, S. N. and Dahm-Daphi, J. Distinct Mechanisms of Nonhomologous End Joining in the Repair of Site-Directed Chromosomal Breaks with Noncomplementary and Complementary Ends. Radiat. Res. 166, 567–574 (2006).DNA double-strand breaks (DSBs) are considered the most important type of DNA damage inflicted by ionizing radiation. The molecular mechanisms of DSB repair by nonhomologous end joining (NHEJ) have not been well studied in live mammalian cells, due in part to the lack of suitable chromosomal repair assays. We previously introduced a novel plasmid-based assay to monitor NHEJ of site-directed chromosomal I-SceI breaks. In the current study, we expanded the analysis of chromosomal NHEJ products in murine fibroblasts to focus on the error-prone rejoining of DSBs with noncomplementary ends, which may serve as a model for radiation damage repair. We found that noncomplementary ends were efficiently repaired using microhomologies of 1–2 nucleotides (nt) present in the single-stranded overhangs, thereby keeping repair-associated end degradation to a minimum (2–3 nt). Microhomology-mediated end joining was disrupted by Wortmannin, a known inhibitor of DNA-PKcs. However, Wortmannin did not significantly impair the proficiency of end joining. In contrast to noncomplementary ends, the rejoining of cohesive ends showed only a minor dependence on microhomologies but produced fivefold larger deletions than the repair of noncomplementary ends. Together, these data suggest the presence of several distinct NHEJ mechanisms in live cells, which are characterized by the degree of sequence deletion and microhomology use. Our NHEJ assay should prove a useful system to further elucidate the genetic determinants and molecular mechanisms of site-directed DSBs in living cells." @default.
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• W2059920795 date "2006-10-01" @default.
• W2059920795 modified "2023-10-16" @default.
• W2059920795 title "Distinct Mechanisms of Nonhomologous End Joining in the Repair of Site-Directed Chromosomal Breaks with Noncomplementary and Complementary Ends" @default.
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• W2059920795 doi "https://doi.org/10.1667/rr0524.1" @default.
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