Single cell analysis of transcriptional activation dynamics.

<h4>Background</h4>Gene activation is thought to occur through a series of temporally defined regulatory steps. However, this process has not been completely evaluated in single living mammalian cells.<h4>Methodology/principal findings</h4>To investigate the timing and coordi...

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Autores principales:	Ilona U Rafalska-Metcalf, Sara Lawrence Powers, Lucy M Joo, Gary LeRoy, Susan M Janicki
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2010
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/528fe1c904134c8f9e7eff32eaf52241
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spelling	oai:doaj.org-article:528fe1c904134c8f9e7eff32eaf522412021-11-25T06:24:23ZSingle cell analysis of transcriptional activation dynamics.1932-620310.1371/journal.pone.0010272https://doaj.org/article/528fe1c904134c8f9e7eff32eaf522412010-04-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/20422051/pdf/?tool=EBIhttps://doaj.org/toc/1932-6203<h4>Background</h4>Gene activation is thought to occur through a series of temporally defined regulatory steps. However, this process has not been completely evaluated in single living mammalian cells.<h4>Methodology/principal findings</h4>To investigate the timing and coordination of gene activation events, we tracked the recruitment of GCN5 (histone acetyltransferase), RNA polymerase II, Brd2 and Brd4 (acetyl-lysine binding proteins), in relation to a VP16-transcriptional activator, to a transcription site that can be visualized in single living cells. All accumulated rapidly with the VP16 activator as did the transcribed RNA. RNA was also detected at significantly more transcription sites in cells expressing the VP16-activator compared to a p53-activator. After alpha-amanitin pre-treatment, the VP16-activator, GCN5, and Brd2 are still recruited to the transcription site but the chromatin does not decondense.<h4>Conclusions/significance</h4>This study demonstrates that a strong activator can rapidly overcome the condensed chromatin structure of an inactive transcription site and supercede the expected requirement for regulatory events to proceed in a temporally defined order. Additionally, activator strength determines the number of cells in which transcription is induced as well as the extent of chromatin decondensation. As chromatin decondensation is significantly reduced after alpha-amanitin pre-treatment, despite the recruitment of transcriptional activation factors, this provides further evidence that transcription drives large-scale chromatin decondensation.Ilona U Rafalska-MetcalfSara Lawrence PowersLucy M JooGary LeRoySusan M JanickiPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 5, Iss 4, p e10272 (2010)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Ilona U Rafalska-Metcalf Sara Lawrence Powers Lucy M Joo Gary LeRoy Susan M Janicki Single cell analysis of transcriptional activation dynamics.
description	<h4>Background</h4>Gene activation is thought to occur through a series of temporally defined regulatory steps. However, this process has not been completely evaluated in single living mammalian cells.<h4>Methodology/principal findings</h4>To investigate the timing and coordination of gene activation events, we tracked the recruitment of GCN5 (histone acetyltransferase), RNA polymerase II, Brd2 and Brd4 (acetyl-lysine binding proteins), in relation to a VP16-transcriptional activator, to a transcription site that can be visualized in single living cells. All accumulated rapidly with the VP16 activator as did the transcribed RNA. RNA was also detected at significantly more transcription sites in cells expressing the VP16-activator compared to a p53-activator. After alpha-amanitin pre-treatment, the VP16-activator, GCN5, and Brd2 are still recruited to the transcription site but the chromatin does not decondense.<h4>Conclusions/significance</h4>This study demonstrates that a strong activator can rapidly overcome the condensed chromatin structure of an inactive transcription site and supercede the expected requirement for regulatory events to proceed in a temporally defined order. Additionally, activator strength determines the number of cells in which transcription is induced as well as the extent of chromatin decondensation. As chromatin decondensation is significantly reduced after alpha-amanitin pre-treatment, despite the recruitment of transcriptional activation factors, this provides further evidence that transcription drives large-scale chromatin decondensation.
format	article
author	Ilona U Rafalska-Metcalf Sara Lawrence Powers Lucy M Joo Gary LeRoy Susan M Janicki
author_facet	Ilona U Rafalska-Metcalf Sara Lawrence Powers Lucy M Joo Gary LeRoy Susan M Janicki
author_sort	Ilona U Rafalska-Metcalf
title	Single cell analysis of transcriptional activation dynamics.
title_short	Single cell analysis of transcriptional activation dynamics.
title_full	Single cell analysis of transcriptional activation dynamics.
title_fullStr	Single cell analysis of transcriptional activation dynamics.
title_full_unstemmed	Single cell analysis of transcriptional activation dynamics.
title_sort	single cell analysis of transcriptional activation dynamics.
publisher	Public Library of Science (PLoS)
publishDate	2010
url	https://doaj.org/article/528fe1c904134c8f9e7eff32eaf52241
work_keys_str_mv	AT ilonaurafalskametcalf singlecellanalysisoftranscriptionalactivationdynamics AT saralawrencepowers singlecellanalysisoftranscriptionalactivationdynamics AT lucymjoo singlecellanalysisoftranscriptionalactivationdynamics AT garyleroy singlecellanalysisoftranscriptionalactivationdynamics AT susanmjanicki singlecellanalysisoftranscriptionalactivationdynamics
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Single cell analysis of transcriptional activation dynamics.

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