Abstract
We have built an ion-microbeam for studies of the nuclear topography and kinetics of double-strand break repair at the single cell level. Here, we show that a first and a second, delayed single ion exposure at different nuclear sites led to comparable accumulations of phospho-ATM, γ-H2AX and Mdc1 at both earlier (e) and later (l) microirradiated sites. In contrast, accumulations of 53BP1 and the recombination protein Rad51 were strongly reduced at l-sites. This apparent competition effect is accompanied by a reduced amount of 53BP1 in undamaged areas of the irradiated nuclei. We suggest that a critically limited pool size combined with strong binding at irradiated sites leads to the exhaustion of unbound factors freely roaming the nuclear space. The undersupply of these factors at l-sites requires in addition a long-lasting binding at e-sites or a weaker binding at l-sites. The observed effects suggest that DNA damage response at individual nuclear sites depends on the time course of damage load. This may have implications for therapeutic radiation treatments.
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Acknowledgments
We thank the staff of the Maier–Leibnitz-Laboratorium for operating the accelerator and F. Eckardt-Schupp, GSF Research Centre Neuherberg, for providing access to the α-irradiation facility. This work was supported by grants of the European Science Foundation (ESF) under the EUROCORES Programme EuroDYNA (G.D., T.C., A.A.F.), the DFG Cluster of Excellence Munich Centre for Advanced Photonics (G.D., A.A.F.), the Bundesamt für Strahlenschutz (A.A.F.) and the Bundesministerium für Bildung und Forschung (A.A.F.).
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Christoph Greubel, Volker Hable and Guido A. Drexler contributed equally to this work.
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Greubel, C., Hable, V., Drexler, G.A. et al. Competition effect in DNA damage response. Radiat Environ Biophys 47, 423–429 (2008). https://doi.org/10.1007/s00411-008-0182-z
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DOI: https://doi.org/10.1007/s00411-008-0182-z