Title |
Identification of the elementary structural units of the DNA damage response
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Published in |
Nature Communications, June 2017
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DOI | 10.1038/ncomms15760 |
Pubmed ID | |
Authors |
Francesco Natale, Alexander Rapp, Wei Yu, Andreas Maiser, Hartmann Harz, Annina Scholl, Stephan Grulich, Tobias Anton, David Hörl, Wei Chen, Marco Durante, Gisela Taucher-Scholz, Heinrich Leonhardt, M. Cristina Cardoso |
Abstract |
Histone H2AX phosphorylation is an early signalling event triggered by DNA double-strand breaks (DSBs). To elucidate the elementary units of phospho-H2AX-labelled chromatin, we integrate super-resolution microscopy of phospho-H2AX during DNA repair in human cells with genome-wide sequencing analyses. Here we identify phospho-H2AX chromatin domains in the nanometre range with median length of ∼75 kb. Correlation analysis with over 60 genomic features shows a time-dependent euchromatin-to-heterochromatin repair trend. After X-ray or CRISPR-Cas9-mediated DSBs, phospho-H2AX-labelled heterochromatin exhibits DNA decondensation while retaining heterochromatic histone marks, indicating that chromatin structural and molecular determinants are uncoupled during repair. The phospho-H2AX nano-domains arrange into higher-order clustered structures of discontinuously phosphorylated chromatin, flanked by CTCF. CTCF knockdown impairs spreading of the phosphorylation throughout the 3D-looped nano-domains. Co-staining of phospho-H2AX with phospho-Ku70 and TUNEL reveals that clusters rather than nano-foci represent single DSBs. Hence, each chromatin loop is a nano-focus, whose clusters correspond to previously known phospho-H2AX foci. |
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United Kingdom | 9 | 32% |
United States | 3 | 11% |
France | 2 | 7% |
Italy | 1 | 4% |
Denmark | 1 | 4% |
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Australia | 1 | 4% |
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Demographic breakdown
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Members of the public | 14 | 50% |
Scientists | 11 | 39% |
Science communicators (journalists, bloggers, editors) | 3 | 11% |
Mendeley readers
Geographical breakdown
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---|---|---|
Netherlands | 1 | <1% |
Unknown | 225 | 100% |
Demographic breakdown
Readers by professional status | Count | As % |
---|---|---|
Student > Ph. D. Student | 54 | 24% |
Researcher | 37 | 16% |
Student > Master | 21 | 9% |
Student > Bachelor | 17 | 8% |
Student > Postgraduate | 14 | 6% |
Other | 33 | 15% |
Unknown | 50 | 22% |
Readers by discipline | Count | As % |
---|---|---|
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Agricultural and Biological Sciences | 25 | 11% |
Physics and Astronomy | 12 | 5% |
Medicine and Dentistry | 7 | 3% |
Chemistry | 6 | 3% |
Other | 18 | 8% |
Unknown | 55 | 24% |