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Identification of the elementary structural units of the DNA damage response

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...

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Autores principales: Natale, Francesco, Rapp, Alexander, Yu, Wei, Maiser, Andreas, Harz, Hartmann, Scholl, Annina, Grulich, Stephan, Anton, Tobias, Hörl, David, Chen, Wei, Durante, Marco, Taucher-Scholz, Gisela, Leonhardt, Heinrich, Cardoso, M. Cristina
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5472794/
https://www.ncbi.nlm.nih.gov/pubmed/28604675
http://dx.doi.org/10.1038/ncomms15760
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author Natale, Francesco
Rapp, Alexander
Yu, Wei
Maiser, Andreas
Harz, Hartmann
Scholl, Annina
Grulich, Stephan
Anton, Tobias
Hörl, David
Chen, Wei
Durante, Marco
Taucher-Scholz, Gisela
Leonhardt, Heinrich
Cardoso, M. Cristina
author_facet Natale, Francesco
Rapp, Alexander
Yu, Wei
Maiser, Andreas
Harz, Hartmann
Scholl, Annina
Grulich, Stephan
Anton, Tobias
Hörl, David
Chen, Wei
Durante, Marco
Taucher-Scholz, Gisela
Leonhardt, Heinrich
Cardoso, M. Cristina
author_sort Natale, Francesco
collection PubMed
description 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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spelling pubmed-54727942017-06-28 Identification of the elementary structural units of the DNA damage response Natale, Francesco Rapp, Alexander Yu, Wei Maiser, Andreas Harz, Hartmann Scholl, Annina Grulich, Stephan Anton, Tobias Hörl, David Chen, Wei Durante, Marco Taucher-Scholz, Gisela Leonhardt, Heinrich Cardoso, M. Cristina Nat Commun Article 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. Nature Publishing Group 2017-06-12 /pmc/articles/PMC5472794/ /pubmed/28604675 http://dx.doi.org/10.1038/ncomms15760 Text en Copyright © 2017, The Author(s) http://creativecommons.org/licenses/by/4.0/ Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/
spellingShingle Article
Natale, Francesco
Rapp, Alexander
Yu, Wei
Maiser, Andreas
Harz, Hartmann
Scholl, Annina
Grulich, Stephan
Anton, Tobias
Hörl, David
Chen, Wei
Durante, Marco
Taucher-Scholz, Gisela
Leonhardt, Heinrich
Cardoso, M. Cristina
Identification of the elementary structural units of the DNA damage response
title Identification of the elementary structural units of the DNA damage response
title_full Identification of the elementary structural units of the DNA damage response
title_fullStr Identification of the elementary structural units of the DNA damage response
title_full_unstemmed Identification of the elementary structural units of the DNA damage response
title_short Identification of the elementary structural units of the DNA damage response
title_sort identification of the elementary structural units of the dna damage response
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5472794/
https://www.ncbi.nlm.nih.gov/pubmed/28604675
http://dx.doi.org/10.1038/ncomms15760
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