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A novel and simple micro-irradiation technique for creating localized DNA double-strand breaks.

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タイトル: A novel and simple micro-irradiation technique for creating localized DNA double-strand breaks.
著者: Suzuki, Keiji / Yamauchi, Motohiro / Oka, Yasuyoshi / Suzuki, Masatoshi / Yamashita, Shunichi
発行日: 2010年 7月 1日
出版者: Oxford University Press
引用: Nucleic acids research, 38(12), pp.e129; 2010
抄録: An ataxia-telangiectasia mutated (ATM)-dependent DNA damage signal is amplified through the interaction of various factors, which are recruited to the chromatin regions with DNA double-strand breaks. Spatial and temporal regulation of such factors is analysed by fluorescence microscopy in combination with laser micro-irradiation. Here we describe a novel and simple technique for micro-irradiation that does not require a laser source. Cells were labelled with BrdU for 48-72 h, covered with porous polycarbonate membranes, and exposed to UVC. All BrdU-labelled cells showed localized foci of phosphorylated ATM, phosphorylated histone H2AX, MDC1 and 53BP1 upon irradiation, showing that these foci were induced irrespective of the cell-cycle phase. They were also detectable in nucleotide excision repair-defective XPA cells labelled with BrdU, indicating that the foci did not reflect an excision repair-related process. Furthermore, an ATM-specific inhibitor significantly attenuated the foci formation, and disappearance of the foci was significantly abrogated in non-homologous end-joining-defective cells. Thus, it can be concluded that micro-irradiation generated DNA double-strand breaks in BrdU-sensitized cells. The present technique should accelerate research in the fields of DNA damage response, DNA repair and DNA recombination, as it provides more chances to perform micro-irradiation experiments without any specific equipment.
URI: http://hdl.handle.net/10069/23875
ISSN: 13624962
DOI: 10.1093/nar/gkq226
PubMed ID: 20385602
権利: © The Author(s) 2010. Published by Oxford University Press. / This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.5), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
資料タイプ: Journal Article
原稿種類: publisher
出現コレクション:110 学術雑誌論文

引用URI : http://hdl.handle.net/10069/23875



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