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Ultra-soft X-ray system for imaging the early cellular responses to X-ray induced DNA damage

The majority of the proteins involved in processing of DNA double-strand breaks (DSBs) accumulate at the damage sites. Real-time imaging and analysis of these processes, triggered by the so-called microirradiation using UV lasers or heavy particle beams, yielded valuable insights into the underlying...

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Autores principales: Kochan, Jakub A, van den Belt, Matthias, von der Lippe, Julia, Desclos, Emilie C B, Steurer, Barbara, Hoebe, Ron A, Scutigliani, Enzo M, Verhoeven, Jan, Stap, Jan, Bosch, Ruben, Rijpkema, Meindert, van Oven, Carel, van Veen, Henk A, Stellingwerf, Irene, Vriend, Lianne E M, Marteijn, Jurgen A, Aten, Jacob A, Krawczyk, Przemek M
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6753493/
https://www.ncbi.nlm.nih.gov/pubmed/31318974
http://dx.doi.org/10.1093/nar/gkz609
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author Kochan, Jakub A
van den Belt, Matthias
von der Lippe, Julia
Desclos, Emilie C B
Steurer, Barbara
Hoebe, Ron A
Scutigliani, Enzo M
Verhoeven, Jan
Stap, Jan
Bosch, Ruben
Rijpkema, Meindert
van Oven, Carel
van Veen, Henk A
Stellingwerf, Irene
Vriend, Lianne E M
Marteijn, Jurgen A
Aten, Jacob A
Krawczyk, Przemek M
author_facet Kochan, Jakub A
van den Belt, Matthias
von der Lippe, Julia
Desclos, Emilie C B
Steurer, Barbara
Hoebe, Ron A
Scutigliani, Enzo M
Verhoeven, Jan
Stap, Jan
Bosch, Ruben
Rijpkema, Meindert
van Oven, Carel
van Veen, Henk A
Stellingwerf, Irene
Vriend, Lianne E M
Marteijn, Jurgen A
Aten, Jacob A
Krawczyk, Przemek M
author_sort Kochan, Jakub A
collection PubMed
description The majority of the proteins involved in processing of DNA double-strand breaks (DSBs) accumulate at the damage sites. Real-time imaging and analysis of these processes, triggered by the so-called microirradiation using UV lasers or heavy particle beams, yielded valuable insights into the underlying DSB repair mechanisms. To study the temporal organization of DSB repair responses triggered by a more clinically-relevant DNA damaging agent, we developed a system coined X-ray multi-microbeam microscope (XM3), capable of simultaneous high dose-rate (micro)irradiation of large numbers of cells with ultra-soft X-rays and imaging of the ensuing cellular responses. Using this setup, we analyzed the changes in real-time kinetics of MRE11, MDC1, RNF8, RNF168 and 53BP1—proteins involved in the signaling axis of mammalian DSB repair—in response to X-ray and UV laser-induced DNA damage, in non-cancerous and cancer cells and in the presence or absence of a photosensitizer. Our results reveal, for the first time, the kinetics of DSB signaling triggered by X-ray microirradiation and establish XM3 as a powerful platform for real-time analysis of cellular DSB repair responses.
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spelling pubmed-67534932019-09-25 Ultra-soft X-ray system for imaging the early cellular responses to X-ray induced DNA damage Kochan, Jakub A van den Belt, Matthias von der Lippe, Julia Desclos, Emilie C B Steurer, Barbara Hoebe, Ron A Scutigliani, Enzo M Verhoeven, Jan Stap, Jan Bosch, Ruben Rijpkema, Meindert van Oven, Carel van Veen, Henk A Stellingwerf, Irene Vriend, Lianne E M Marteijn, Jurgen A Aten, Jacob A Krawczyk, Przemek M Nucleic Acids Res Methods Online The majority of the proteins involved in processing of DNA double-strand breaks (DSBs) accumulate at the damage sites. Real-time imaging and analysis of these processes, triggered by the so-called microirradiation using UV lasers or heavy particle beams, yielded valuable insights into the underlying DSB repair mechanisms. To study the temporal organization of DSB repair responses triggered by a more clinically-relevant DNA damaging agent, we developed a system coined X-ray multi-microbeam microscope (XM3), capable of simultaneous high dose-rate (micro)irradiation of large numbers of cells with ultra-soft X-rays and imaging of the ensuing cellular responses. Using this setup, we analyzed the changes in real-time kinetics of MRE11, MDC1, RNF8, RNF168 and 53BP1—proteins involved in the signaling axis of mammalian DSB repair—in response to X-ray and UV laser-induced DNA damage, in non-cancerous and cancer cells and in the presence or absence of a photosensitizer. Our results reveal, for the first time, the kinetics of DSB signaling triggered by X-ray microirradiation and establish XM3 as a powerful platform for real-time analysis of cellular DSB repair responses. Oxford University Press 2019-09-26 2019-07-18 /pmc/articles/PMC6753493/ /pubmed/31318974 http://dx.doi.org/10.1093/nar/gkz609 Text en © The Author(s) 2019. Published by Oxford University Press on behalf of Nucleic Acids Research. http://creativecommons.org/licenses/by-nc/4.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com
spellingShingle Methods Online
Kochan, Jakub A
van den Belt, Matthias
von der Lippe, Julia
Desclos, Emilie C B
Steurer, Barbara
Hoebe, Ron A
Scutigliani, Enzo M
Verhoeven, Jan
Stap, Jan
Bosch, Ruben
Rijpkema, Meindert
van Oven, Carel
van Veen, Henk A
Stellingwerf, Irene
Vriend, Lianne E M
Marteijn, Jurgen A
Aten, Jacob A
Krawczyk, Przemek M
Ultra-soft X-ray system for imaging the early cellular responses to X-ray induced DNA damage
title Ultra-soft X-ray system for imaging the early cellular responses to X-ray induced DNA damage
title_full Ultra-soft X-ray system for imaging the early cellular responses to X-ray induced DNA damage
title_fullStr Ultra-soft X-ray system for imaging the early cellular responses to X-ray induced DNA damage
title_full_unstemmed Ultra-soft X-ray system for imaging the early cellular responses to X-ray induced DNA damage
title_short Ultra-soft X-ray system for imaging the early cellular responses to X-ray induced DNA damage
title_sort ultra-soft x-ray system for imaging the early cellular responses to x-ray induced dna damage
topic Methods Online
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6753493/
https://www.ncbi.nlm.nih.gov/pubmed/31318974
http://dx.doi.org/10.1093/nar/gkz609
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