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Live-cell imaging reveals the dynamics of PRC2 and recruitment to chromatin by SUZ12-associated subunits

Polycomb-repressive complex 2 (PRC2) is a histone methyltransferase that promotes epigenetic gene silencing, but the dynamics of its interactions with chromatin are largely unknown. Here we quantitatively measured the binding of PRC2 to chromatin in human cancer cells. Genome editing of a HaloTag in...

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Detalles Bibliográficos
Autores principales: Youmans, Daniel T., Schmidt, Jens C., Cech, Thomas R.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Cold Spring Harbor Laboratory Press 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6049511/
https://www.ncbi.nlm.nih.gov/pubmed/29891558
http://dx.doi.org/10.1101/gad.311936.118
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author Youmans, Daniel T.
Schmidt, Jens C.
Cech, Thomas R.
author_facet Youmans, Daniel T.
Schmidt, Jens C.
Cech, Thomas R.
author_sort Youmans, Daniel T.
collection PubMed
description Polycomb-repressive complex 2 (PRC2) is a histone methyltransferase that promotes epigenetic gene silencing, but the dynamics of its interactions with chromatin are largely unknown. Here we quantitatively measured the binding of PRC2 to chromatin in human cancer cells. Genome editing of a HaloTag into the endogenous EZH2 and SUZ12 loci and single-particle tracking revealed that ∼80% of PRC2 rapidly diffuses through the nucleus, while ∼20% is chromatin-bound. Short-term treatment with a small molecule inhibitor of the EED–H3K27me3 interaction had no immediate effect on the chromatin residence time of PRC2. In contrast, separation-of-function mutants of SUZ12, which still form the core PRC2 complex but cannot bind accessory proteins, revealed a major contribution of AEBP2 and PCL homolog proteins to chromatin binding. We therefore quantified the dynamics of this chromatin-modifying complex in living cells and separated the contributions of H3K27me3 histone marks and various PRC2 subunits to recruitment of PRC2 to chromatin.
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spelling pubmed-60495112018-12-01 Live-cell imaging reveals the dynamics of PRC2 and recruitment to chromatin by SUZ12-associated subunits Youmans, Daniel T. Schmidt, Jens C. Cech, Thomas R. Genes Dev Research Paper Polycomb-repressive complex 2 (PRC2) is a histone methyltransferase that promotes epigenetic gene silencing, but the dynamics of its interactions with chromatin are largely unknown. Here we quantitatively measured the binding of PRC2 to chromatin in human cancer cells. Genome editing of a HaloTag into the endogenous EZH2 and SUZ12 loci and single-particle tracking revealed that ∼80% of PRC2 rapidly diffuses through the nucleus, while ∼20% is chromatin-bound. Short-term treatment with a small molecule inhibitor of the EED–H3K27me3 interaction had no immediate effect on the chromatin residence time of PRC2. In contrast, separation-of-function mutants of SUZ12, which still form the core PRC2 complex but cannot bind accessory proteins, revealed a major contribution of AEBP2 and PCL homolog proteins to chromatin binding. We therefore quantified the dynamics of this chromatin-modifying complex in living cells and separated the contributions of H3K27me3 histone marks and various PRC2 subunits to recruitment of PRC2 to chromatin. Cold Spring Harbor Laboratory Press 2018-06-01 /pmc/articles/PMC6049511/ /pubmed/29891558 http://dx.doi.org/10.1101/gad.311936.118 Text en © 2018 Youmans et al.; Published by Cold Spring Harbor Laboratory Press http://creativecommons.org/licenses/by-nc/4.0/ This article is distributed exclusively by Cold Spring Harbor Laboratory Press for the first six months after the full-issue publication date (see http://genesdev.cshlp.org/site/misc/terms.xhtml). After six months, it is available under a Creative Commons License (Attribution-NonCommercial 4.0 International), as described at http://creativecommons.org/licenses/by-nc/4.0/.
spellingShingle Research Paper
Youmans, Daniel T.
Schmidt, Jens C.
Cech, Thomas R.
Live-cell imaging reveals the dynamics of PRC2 and recruitment to chromatin by SUZ12-associated subunits
title Live-cell imaging reveals the dynamics of PRC2 and recruitment to chromatin by SUZ12-associated subunits
title_full Live-cell imaging reveals the dynamics of PRC2 and recruitment to chromatin by SUZ12-associated subunits
title_fullStr Live-cell imaging reveals the dynamics of PRC2 and recruitment to chromatin by SUZ12-associated subunits
title_full_unstemmed Live-cell imaging reveals the dynamics of PRC2 and recruitment to chromatin by SUZ12-associated subunits
title_short Live-cell imaging reveals the dynamics of PRC2 and recruitment to chromatin by SUZ12-associated subunits
title_sort live-cell imaging reveals the dynamics of prc2 and recruitment to chromatin by suz12-associated subunits
topic Research Paper
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6049511/
https://www.ncbi.nlm.nih.gov/pubmed/29891558
http://dx.doi.org/10.1101/gad.311936.118
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