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Live-cell imaging uncovers the relationship between histone acetylation, transcription initiation, and nucleosome mobility

Histone acetylation and RNA polymerase II phosphorylation are associated with transcriptionally active chromatin, but their spatiotemporal relationship in live cells remains poorly understood. To address this problem, we combine Fab-based labeling of endogenous protein modifications with single-mole...

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Autores principales: Saxton, Matthew N., Morisaki, Tatsuya, Krapf, Diego, Kimura, Hiroshi, Stasevich, Timothy J.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Association for the Advancement of Science 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10550241/
https://www.ncbi.nlm.nih.gov/pubmed/37792937
http://dx.doi.org/10.1126/sciadv.adh4819
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author Saxton, Matthew N.
Morisaki, Tatsuya
Krapf, Diego
Kimura, Hiroshi
Stasevich, Timothy J.
author_facet Saxton, Matthew N.
Morisaki, Tatsuya
Krapf, Diego
Kimura, Hiroshi
Stasevich, Timothy J.
author_sort Saxton, Matthew N.
collection PubMed
description Histone acetylation and RNA polymerase II phosphorylation are associated with transcriptionally active chromatin, but their spatiotemporal relationship in live cells remains poorly understood. To address this problem, we combine Fab-based labeling of endogenous protein modifications with single-molecule tracking to quantify the dynamics of chromatin enriched with histone H3 lysine-27 acetylation (H3K27ac) and RNA polymerase II serine-5 phosphorylation (RNAP2-Ser5ph). Our analysis reveals that chromatin enriched with these two modifications is generally separate. In these separated sites, we show that the two modifications are inversely correlated with one another on the minutes time scale and that single nucleosomes within each region display distinct and opposing dynamics on the subsecond time scale. While nucleosomes diffuse ~15% faster in chromatin enriched with H3K27ac, they diffuse ~15% slower in chromatin enriched with RNAP2-Ser5ph. These results argue that high levels of H3K27ac and RNAP2-Ser5ph are not often present together at the same place and time, but rather each marks distinct transcriptionally poised or active sites, respectively.
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spelling pubmed-105502412023-10-05 Live-cell imaging uncovers the relationship between histone acetylation, transcription initiation, and nucleosome mobility Saxton, Matthew N. Morisaki, Tatsuya Krapf, Diego Kimura, Hiroshi Stasevich, Timothy J. Sci Adv Biomedicine and Life Sciences Histone acetylation and RNA polymerase II phosphorylation are associated with transcriptionally active chromatin, but their spatiotemporal relationship in live cells remains poorly understood. To address this problem, we combine Fab-based labeling of endogenous protein modifications with single-molecule tracking to quantify the dynamics of chromatin enriched with histone H3 lysine-27 acetylation (H3K27ac) and RNA polymerase II serine-5 phosphorylation (RNAP2-Ser5ph). Our analysis reveals that chromatin enriched with these two modifications is generally separate. In these separated sites, we show that the two modifications are inversely correlated with one another on the minutes time scale and that single nucleosomes within each region display distinct and opposing dynamics on the subsecond time scale. While nucleosomes diffuse ~15% faster in chromatin enriched with H3K27ac, they diffuse ~15% slower in chromatin enriched with RNAP2-Ser5ph. These results argue that high levels of H3K27ac and RNAP2-Ser5ph are not often present together at the same place and time, but rather each marks distinct transcriptionally poised or active sites, respectively. American Association for the Advancement of Science 2023-10-04 /pmc/articles/PMC10550241/ /pubmed/37792937 http://dx.doi.org/10.1126/sciadv.adh4819 Text en Copyright © 2023 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works. Distributed under a Creative Commons Attribution License 4.0 (CC BY). https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution license (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Biomedicine and Life Sciences
Saxton, Matthew N.
Morisaki, Tatsuya
Krapf, Diego
Kimura, Hiroshi
Stasevich, Timothy J.
Live-cell imaging uncovers the relationship between histone acetylation, transcription initiation, and nucleosome mobility
title Live-cell imaging uncovers the relationship between histone acetylation, transcription initiation, and nucleosome mobility
title_full Live-cell imaging uncovers the relationship between histone acetylation, transcription initiation, and nucleosome mobility
title_fullStr Live-cell imaging uncovers the relationship between histone acetylation, transcription initiation, and nucleosome mobility
title_full_unstemmed Live-cell imaging uncovers the relationship between histone acetylation, transcription initiation, and nucleosome mobility
title_short Live-cell imaging uncovers the relationship between histone acetylation, transcription initiation, and nucleosome mobility
title_sort live-cell imaging uncovers the relationship between histone acetylation, transcription initiation, and nucleosome mobility
topic Biomedicine and Life Sciences
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10550241/
https://www.ncbi.nlm.nih.gov/pubmed/37792937
http://dx.doi.org/10.1126/sciadv.adh4819
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