TY - JOUR
T1 - Ultra-soft X-ray system for imaging the early cellular responses to X-ray induced DNA damage
AU - Kochan, Jakub A.
AU - van den Belt, Matthias
AU - von der Lippe, Julia
AU - Desclos, Emilie C. B.
AU - Steurer, Barbara
AU - Hoebe, Ron A.
AU - Scutigliani, Enzo M.
AU - Verhoeven, Jan
AU - Stap, Jan
AU - Bosch, Ruben
AU - Rijpkema, Meindert
AU - van Oven, Carel
AU - van Veen, Henk A.
AU - Stellingwerf, Irene
AU - Vriend, Lianne E. M.
AU - Marteijn, Jurgen A.
AU - Aten, Jacob A.
AU - Krawczyk, Przemek M.
PY - 2019
Y1 - 2019
N2 - 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.
AB - 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.
UR - https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=85072509741&origin=inward
UR - https://www.ncbi.nlm.nih.gov/pubmed/31318974
U2 - https://doi.org/10.1093/nar/gkz609
DO - https://doi.org/10.1093/nar/gkz609
M3 - Article
C2 - 31318974
SN - 0305-1048
VL - 47
SP - e100
JO - Nucleic Acids Research
JF - Nucleic Acids Research
IS - 17
ER -