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

Jakub A. Kochan, Matthias van den Belt, Julia von der Lippe, Emilie C. B. Desclos, Barbara Steurer, Ron A. Hoebe, Enzo M. Scutigliani, Jan Verhoeven, Jan Stap, Ruben Bosch, Meindert Rijpkema, Carel van Oven, Henk A. van Veen, Irene Stellingwerf, Lianne E. M. Vriend, Jurgen A. Marteijn, Jacob A. Aten, Przemek M. Krawczyk

Research output: Contribution to journalArticleAcademicpeer-review

9 Citations (Scopus)

Abstract

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.
Original languageEnglish
Pages (from-to)e100
JournalNucleic Acids Research
Volume47
Issue number17
DOIs
Publication statusPublished - 2019

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