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Identification of the elementary structural units of the DNA damage response.

Natale, Francesco and Rapp, Alexander and Yu, Wei and Maiser, Andreas and Harz, Hartmann and Scholl, Annina and Grulich, Stephan and Anton, Tobias and Hörl, David and Chen, Wei and Durante, Marco and Taucher-Scholz, Gisela and Leonhardt, Heinrich and Cardoso, M. Cristina (2017):
Identification of the elementary structural units of the DNA damage response.
In: Nature communications, p. 15760, 8, ISSN 2041-1723, [Article]

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.

Item Type: Article
Erschienen: 2017
Creators: Natale, Francesco and Rapp, Alexander and Yu, Wei and Maiser, Andreas and Harz, Hartmann and Scholl, Annina and Grulich, Stephan and Anton, Tobias and Hörl, David and Chen, Wei and Durante, Marco and Taucher-Scholz, Gisela and Leonhardt, Heinrich and Cardoso, M. Cristina
Title: Identification of the elementary structural units of the DNA damage response.
Language: English
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.

Journal or Publication Title: Nature communications
Volume: 8
Divisions: 10 Department of Biology
10 Department of Biology > Cell Biology and Epigenetics
Date Deposited: 20 Jun 2017 06:40
Identification Number: pmid:28604675
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