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Low level phosphorylation of histone H2AX on serine 139 (γH2AX) is not associated with DNA double-strand breaks

Phosphorylation of histone H2AX on serine 139 (γH2AX) is an early step in cellular response to a DNA double-strand break (DSB). γH2AX foci are generally regarded as markers of DSBs. A growing body of evidence demonstrates, however, that while induction of DSBs always brings about phosphorylation of...

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Autores principales: Rybak, Paulina, Hoang, Agnieszka, Bujnowicz, Lukasz, Bernas, Tytus, Berniak, Krzysztof, Zarębski, Mirosław, Darzynkiewicz, Zbigniew, Dobrucki, Jerzy
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Impact Journals LLC 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5226530/
https://www.ncbi.nlm.nih.gov/pubmed/27391338
http://dx.doi.org/10.18632/oncotarget.10411
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author Rybak, Paulina
Hoang, Agnieszka
Bujnowicz, Lukasz
Bernas, Tytus
Berniak, Krzysztof
Zarębski, Mirosław
Darzynkiewicz, Zbigniew
Dobrucki, Jerzy
author_facet Rybak, Paulina
Hoang, Agnieszka
Bujnowicz, Lukasz
Bernas, Tytus
Berniak, Krzysztof
Zarębski, Mirosław
Darzynkiewicz, Zbigniew
Dobrucki, Jerzy
author_sort Rybak, Paulina
collection PubMed
description Phosphorylation of histone H2AX on serine 139 (γH2AX) is an early step in cellular response to a DNA double-strand break (DSB). γH2AX foci are generally regarded as markers of DSBs. A growing body of evidence demonstrates, however, that while induction of DSBs always brings about phosphorylation of histone H2AX, the reverse is not true - the presence of γH2AX foci should not be considered an unequivocal marker of DNA double-strand breaks. We studied DNA damage induced in A549 human lung adenocarcinoma cells by topoisomerase type I and II inhibitors (0.2 μM camptothecin, 10 μM etoposide or 0.2 μM mitoxantrone for 1 h), and using 3D high resolution quantitative confocal microscopy, assessed the number, size and the integrated intensity of immunofluorescence signals of individual γH2AX foci induced by these drugs. Also, investigated was spatial association between γH2AX foci and foci of 53BP1, the protein involved in DSB repair, both in relation to DNA replication sites (factories) as revealed by labeling nascent DNA with EdU. Extensive 3D and correlation data analysis demonstrated that γH2AX foci exhibit a wide range of sizes and levels of H2AX phosphorylation, and correlate differently with 53BP1 and DNA replication. This is the first report showing lack of a link between low level phosphorylation γH2AX sites and double-strand DNA breaks in cells exposed to topoisomerase I or II inhibitors. The data are discussed in terms of mechanisms that may be involved in formation of γH2AX sites of different sizes and intensities.
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spelling pubmed-52265302017-01-18 Low level phosphorylation of histone H2AX on serine 139 (γH2AX) is not associated with DNA double-strand breaks Rybak, Paulina Hoang, Agnieszka Bujnowicz, Lukasz Bernas, Tytus Berniak, Krzysztof Zarębski, Mirosław Darzynkiewicz, Zbigniew Dobrucki, Jerzy Oncotarget Research Paper Phosphorylation of histone H2AX on serine 139 (γH2AX) is an early step in cellular response to a DNA double-strand break (DSB). γH2AX foci are generally regarded as markers of DSBs. A growing body of evidence demonstrates, however, that while induction of DSBs always brings about phosphorylation of histone H2AX, the reverse is not true - the presence of γH2AX foci should not be considered an unequivocal marker of DNA double-strand breaks. We studied DNA damage induced in A549 human lung adenocarcinoma cells by topoisomerase type I and II inhibitors (0.2 μM camptothecin, 10 μM etoposide or 0.2 μM mitoxantrone for 1 h), and using 3D high resolution quantitative confocal microscopy, assessed the number, size and the integrated intensity of immunofluorescence signals of individual γH2AX foci induced by these drugs. Also, investigated was spatial association between γH2AX foci and foci of 53BP1, the protein involved in DSB repair, both in relation to DNA replication sites (factories) as revealed by labeling nascent DNA with EdU. Extensive 3D and correlation data analysis demonstrated that γH2AX foci exhibit a wide range of sizes and levels of H2AX phosphorylation, and correlate differently with 53BP1 and DNA replication. This is the first report showing lack of a link between low level phosphorylation γH2AX sites and double-strand DNA breaks in cells exposed to topoisomerase I or II inhibitors. The data are discussed in terms of mechanisms that may be involved in formation of γH2AX sites of different sizes and intensities. Impact Journals LLC 2016-07-06 /pmc/articles/PMC5226530/ /pubmed/27391338 http://dx.doi.org/10.18632/oncotarget.10411 Text en Copyright: © 2016 Rybak et al. http://creativecommons.org/licenses/by/2.5/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Paper
Rybak, Paulina
Hoang, Agnieszka
Bujnowicz, Lukasz
Bernas, Tytus
Berniak, Krzysztof
Zarębski, Mirosław
Darzynkiewicz, Zbigniew
Dobrucki, Jerzy
Low level phosphorylation of histone H2AX on serine 139 (γH2AX) is not associated with DNA double-strand breaks
title Low level phosphorylation of histone H2AX on serine 139 (γH2AX) is not associated with DNA double-strand breaks
title_full Low level phosphorylation of histone H2AX on serine 139 (γH2AX) is not associated with DNA double-strand breaks
title_fullStr Low level phosphorylation of histone H2AX on serine 139 (γH2AX) is not associated with DNA double-strand breaks
title_full_unstemmed Low level phosphorylation of histone H2AX on serine 139 (γH2AX) is not associated with DNA double-strand breaks
title_short Low level phosphorylation of histone H2AX on serine 139 (γH2AX) is not associated with DNA double-strand breaks
title_sort low level phosphorylation of histone h2ax on serine 139 (γh2ax) is not associated with dna double-strand breaks
topic Research Paper
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5226530/
https://www.ncbi.nlm.nih.gov/pubmed/27391338
http://dx.doi.org/10.18632/oncotarget.10411
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