Comparison of High- and Low-LET Radiation-Induced DNA Double-Strand Break Processing in Living Cells
High-linear-energy-transfer (LET) radiation is more lethal than similar doses of low-LET radiation types, probably a result of the condensed energy deposition pattern of high-LET radiation. Here, we compare high-LET α-particle to low-LET X-ray irradiation and monitor double-strand break (DSB) proces...
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MDPI AG
2020-09-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/21/18/6602 |
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| author | Stefan J. Roobol Irene van den Bent Wiggert A. van Cappellen Tsion E. Abraham Maarten W. Paul Roland Kanaar Adriaan B. Houtsmuller Dik C. van Gent Jeroen Essers |
| author_facet | Stefan J. Roobol Irene van den Bent Wiggert A. van Cappellen Tsion E. Abraham Maarten W. Paul Roland Kanaar Adriaan B. Houtsmuller Dik C. van Gent Jeroen Essers |
| author_sort | Stefan J. Roobol |
| collection | DOAJ |
| description | High-linear-energy-transfer (LET) radiation is more lethal than similar doses of low-LET radiation types, probably a result of the condensed energy deposition pattern of high-LET radiation. Here, we compare high-LET α-particle to low-LET X-ray irradiation and monitor double-strand break (DSB) processing. Live-cell microscopy was used to monitor DNA double-strand breaks (DSBs), marked by p53-binding protein 1 (53BP1). In addition, the accumulation of the endogenous 53BP1 and replication protein A (RPA) DSB processing proteins was analyzed by immunofluorescence. In contrast to α-particle-induced 53BP1 foci, X-ray-induced foci were resolved quickly and more dynamically as they showed an increase in 53BP1 protein accumulation and size. In addition, the number of individual 53BP1 and RPA foci was higher after X-ray irradiation, while focus intensity was higher after α-particle irradiation. Interestingly, 53BP1 foci induced by α-particles contained multiple RPA foci, suggesting multiple individual resection events, which was not observed after X-ray irradiation. We conclude that high-LET α-particles cause closely interspaced DSBs leading to high local concentrations of repair proteins. Our results point toward a change in DNA damage processing toward DNA end-resection and homologous recombination, possibly due to the depletion of soluble protein in the nucleoplasm. The combination of closely interspaced DSBs and perturbed DNA damage processing could be an explanation for the increased relative biological effectiveness (RBE) of high-LET α-particles compared to X-ray irradiation. |
| first_indexed | 2024-03-10T16:27:11Z |
| format | Article |
| id | doaj.art-2c006943388f4365b3296c835c397b38 |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-10T16:27:11Z |
| publishDate | 2020-09-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-2c006943388f4365b3296c835c397b382023-11-20T13:08:51ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672020-09-012118660210.3390/ijms21186602Comparison of High- and Low-LET Radiation-Induced DNA Double-Strand Break Processing in Living CellsStefan J. Roobol0Irene van den Bent1Wiggert A. van Cappellen2Tsion E. Abraham3Maarten W. Paul4Roland Kanaar5Adriaan B. Houtsmuller6Dik C. van Gent7Jeroen Essers8Department of Molecular Genetics, Erasmus University Medical Center, 3015 GD Rotterdam, The NetherlandsDepartment of Molecular Genetics, Erasmus University Medical Center, 3015 GD Rotterdam, The NetherlandsOptical Imaging Center (OIC), Erasmus University Medical Center, 3015 GD Rotterdam, The NetherlandsOptical Imaging Center (OIC), Erasmus University Medical Center, 3015 GD Rotterdam, The NetherlandsDepartment of Molecular Genetics, Erasmus University Medical Center, 3015 GD Rotterdam, The NetherlandsDepartment of Molecular Genetics, Erasmus University Medical Center, 3015 GD Rotterdam, The NetherlandsOptical Imaging Center (OIC), Erasmus University Medical Center, 3015 GD Rotterdam, The NetherlandsDepartment of Molecular Genetics, Erasmus University Medical Center, 3015 GD Rotterdam, The NetherlandsDepartment of Molecular Genetics, Erasmus University Medical Center, 3015 GD Rotterdam, The NetherlandsHigh-linear-energy-transfer (LET) radiation is more lethal than similar doses of low-LET radiation types, probably a result of the condensed energy deposition pattern of high-LET radiation. Here, we compare high-LET α-particle to low-LET X-ray irradiation and monitor double-strand break (DSB) processing. Live-cell microscopy was used to monitor DNA double-strand breaks (DSBs), marked by p53-binding protein 1 (53BP1). In addition, the accumulation of the endogenous 53BP1 and replication protein A (RPA) DSB processing proteins was analyzed by immunofluorescence. In contrast to α-particle-induced 53BP1 foci, X-ray-induced foci were resolved quickly and more dynamically as they showed an increase in 53BP1 protein accumulation and size. In addition, the number of individual 53BP1 and RPA foci was higher after X-ray irradiation, while focus intensity was higher after α-particle irradiation. Interestingly, 53BP1 foci induced by α-particles contained multiple RPA foci, suggesting multiple individual resection events, which was not observed after X-ray irradiation. We conclude that high-LET α-particles cause closely interspaced DSBs leading to high local concentrations of repair proteins. Our results point toward a change in DNA damage processing toward DNA end-resection and homologous recombination, possibly due to the depletion of soluble protein in the nucleoplasm. The combination of closely interspaced DSBs and perturbed DNA damage processing could be an explanation for the increased relative biological effectiveness (RBE) of high-LET α-particles compared to X-ray irradiation.https://www.mdpi.com/1422-0067/21/18/6602DNA double-strand breakshigh linear energy transferalpha particleshomologous recombinationlive-cell microscopynonhomologous DNA end-joining |
| spellingShingle | Stefan J. Roobol Irene van den Bent Wiggert A. van Cappellen Tsion E. Abraham Maarten W. Paul Roland Kanaar Adriaan B. Houtsmuller Dik C. van Gent Jeroen Essers Comparison of High- and Low-LET Radiation-Induced DNA Double-Strand Break Processing in Living Cells International Journal of Molecular Sciences DNA double-strand breaks high linear energy transfer alpha particles homologous recombination live-cell microscopy nonhomologous DNA end-joining |
| title | Comparison of High- and Low-LET Radiation-Induced DNA Double-Strand Break Processing in Living Cells |
| title_full | Comparison of High- and Low-LET Radiation-Induced DNA Double-Strand Break Processing in Living Cells |
| title_fullStr | Comparison of High- and Low-LET Radiation-Induced DNA Double-Strand Break Processing in Living Cells |
| title_full_unstemmed | Comparison of High- and Low-LET Radiation-Induced DNA Double-Strand Break Processing in Living Cells |
| title_short | Comparison of High- and Low-LET Radiation-Induced DNA Double-Strand Break Processing in Living Cells |
| title_sort | comparison of high and low let radiation induced dna double strand break processing in living cells |
| topic | DNA double-strand breaks high linear energy transfer alpha particles homologous recombination live-cell microscopy nonhomologous DNA end-joining |
| url | https://www.mdpi.com/1422-0067/21/18/6602 |
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