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Imaging the Response to DNA Damage in Heterochromatin Domains

The eukaryotic genome is assembled in a nucleoprotein complex called chromatin, whose organization markedly influences the repair of DNA lesions. For instance, compact chromatin states, broadly categorized as heterochromatin, present a challenging environment for DNA damage repair. Through transcrip...

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Autores principales: Chansard, Audrey, Pobega, Enrico, Caron, Pierre, Polo, Sophie E.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9201110/
https://www.ncbi.nlm.nih.gov/pubmed/35721488
http://dx.doi.org/10.3389/fcell.2022.920267
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author Chansard, Audrey
Pobega, Enrico
Caron, Pierre
Polo, Sophie E.
author_facet Chansard, Audrey
Pobega, Enrico
Caron, Pierre
Polo, Sophie E.
author_sort Chansard, Audrey
collection PubMed
description The eukaryotic genome is assembled in a nucleoprotein complex called chromatin, whose organization markedly influences the repair of DNA lesions. For instance, compact chromatin states, broadly categorized as heterochromatin, present a challenging environment for DNA damage repair. Through transcriptional silencing, heterochromatin also plays a vital role in the maintenance of genomic integrity and cellular homeostasis. It is thus of critical importance to decipher whether and how heterochromatin affects the DNA damage response (DDR) to understand how this chromatin state is preserved after DNA damage. Here, we present two laser micro-irradiation-based methods for imaging the DDR in heterochromatin domains in mammalian cells. These methods allow DNA damage targeting to specific subnuclear compartments, direct visualization of the DDR and image-based quantification of the repair response. We apply them to study DNA double-strand break repair pathways in facultative heterochromatin and the repair of UV photoproducts in constitutive heterochromatin. We discuss the advantages and limitations of these methods compared to other targeted approaches for DNA damage induction.
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spelling pubmed-92011102022-06-17 Imaging the Response to DNA Damage in Heterochromatin Domains Chansard, Audrey Pobega, Enrico Caron, Pierre Polo, Sophie E. Front Cell Dev Biol Cell and Developmental Biology The eukaryotic genome is assembled in a nucleoprotein complex called chromatin, whose organization markedly influences the repair of DNA lesions. For instance, compact chromatin states, broadly categorized as heterochromatin, present a challenging environment for DNA damage repair. Through transcriptional silencing, heterochromatin also plays a vital role in the maintenance of genomic integrity and cellular homeostasis. It is thus of critical importance to decipher whether and how heterochromatin affects the DNA damage response (DDR) to understand how this chromatin state is preserved after DNA damage. Here, we present two laser micro-irradiation-based methods for imaging the DDR in heterochromatin domains in mammalian cells. These methods allow DNA damage targeting to specific subnuclear compartments, direct visualization of the DDR and image-based quantification of the repair response. We apply them to study DNA double-strand break repair pathways in facultative heterochromatin and the repair of UV photoproducts in constitutive heterochromatin. We discuss the advantages and limitations of these methods compared to other targeted approaches for DNA damage induction. Frontiers Media S.A. 2022-06-02 /pmc/articles/PMC9201110/ /pubmed/35721488 http://dx.doi.org/10.3389/fcell.2022.920267 Text en Copyright © 2022 Chansard, Pobega, Caron and Polo. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Cell and Developmental Biology
Chansard, Audrey
Pobega, Enrico
Caron, Pierre
Polo, Sophie E.
Imaging the Response to DNA Damage in Heterochromatin Domains
title Imaging the Response to DNA Damage in Heterochromatin Domains
title_full Imaging the Response to DNA Damage in Heterochromatin Domains
title_fullStr Imaging the Response to DNA Damage in Heterochromatin Domains
title_full_unstemmed Imaging the Response to DNA Damage in Heterochromatin Domains
title_short Imaging the Response to DNA Damage in Heterochromatin Domains
title_sort imaging the response to dna damage in heterochromatin domains
topic Cell and Developmental Biology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9201110/
https://www.ncbi.nlm.nih.gov/pubmed/35721488
http://dx.doi.org/10.3389/fcell.2022.920267
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