Cargando…
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...
Autores principales: | , , |
---|---|
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 |
_version_ | 1783340229364547584 |
---|---|
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. |
format | Online Article Text |
id | pubmed-6049511 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | Cold Spring Harbor Laboratory Press |
record_format | MEDLINE/PubMed |
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 |
work_keys_str_mv | AT youmansdanielt livecellimagingrevealsthedynamicsofprc2andrecruitmenttochromatinbysuz12associatedsubunits AT schmidtjensc livecellimagingrevealsthedynamicsofprc2andrecruitmenttochromatinbysuz12associatedsubunits AT cechthomasr livecellimagingrevealsthedynamicsofprc2andrecruitmenttochromatinbysuz12associatedsubunits |