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Sumoylation of the human histone H4 tail inhibits p300-mediated transcription by RNA polymerase II in cellular extracts
The post-translational modification of histones by the small ubiquitin-like modifier (SUMO) protein has been associated with gene regulation, centromeric localization, and double-strand break repair in eukaryotes. Although sumoylation of histone H4 was specifically associated with gene repression, t...
Autores principales: | , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
eLife Sciences Publications, Ltd
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8626089/ https://www.ncbi.nlm.nih.gov/pubmed/34747692 http://dx.doi.org/10.7554/eLife.67952 |
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author | Leonen, Calvin Jon A Shimada, Miho Weller, Caroline E Nakadai, Tomoyoshi Hsu, Peter L Tyson, Elizabeth L Mishra, Arpit Shelton, Patrick MM Sadilek, Martin Hawkins, R David Zheng, Ning Roeder, Robert G Chatterjee, Champak |
author_facet | Leonen, Calvin Jon A Shimada, Miho Weller, Caroline E Nakadai, Tomoyoshi Hsu, Peter L Tyson, Elizabeth L Mishra, Arpit Shelton, Patrick MM Sadilek, Martin Hawkins, R David Zheng, Ning Roeder, Robert G Chatterjee, Champak |
author_sort | Leonen, Calvin Jon A |
collection | PubMed |
description | The post-translational modification of histones by the small ubiquitin-like modifier (SUMO) protein has been associated with gene regulation, centromeric localization, and double-strand break repair in eukaryotes. Although sumoylation of histone H4 was specifically associated with gene repression, this could not be proven due to the challenge of site-specifically sumoylating H4 in cells. Biochemical crosstalk between SUMO and other histone modifications, such as H4 acetylation and H3 methylation, that are associated with active genes also remains unclear. We addressed these challenges in mechanistic studies using an H4 chemically modified at Lys12 by SUMO-3 (H4K12su) and incorporated into mononucleosomes and chromatinized plasmids for functional studies. Mononucleosome-based assays revealed that H4K12su inhibits transcription-activating H4 tail acetylation by the histone acetyltransferase p300, as well as transcription-associated H3K4 methylation by the extended catalytic module of the Set1/COMPASS (complex of proteins associated with Set1) histone methyltransferase complex. Activator- and p300-dependent in vitro transcription assays with chromatinized plasmids revealed that H4K12su inhibits both H4 tail acetylation and RNA polymerase II-mediated transcription. Finally, cell-based assays with a SUMO-H4 fusion that mimics H4 tail sumoylation confirmed the negative crosstalk between histone sumoylation and acetylation/methylation. Thus, our studies establish the key role for histone sumoylation in gene silencing and its negative biochemical crosstalk with active transcription-associated marks in human cells. |
format | Online Article Text |
id | pubmed-8626089 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | eLife Sciences Publications, Ltd |
record_format | MEDLINE/PubMed |
spelling | pubmed-86260892021-11-29 Sumoylation of the human histone H4 tail inhibits p300-mediated transcription by RNA polymerase II in cellular extracts Leonen, Calvin Jon A Shimada, Miho Weller, Caroline E Nakadai, Tomoyoshi Hsu, Peter L Tyson, Elizabeth L Mishra, Arpit Shelton, Patrick MM Sadilek, Martin Hawkins, R David Zheng, Ning Roeder, Robert G Chatterjee, Champak eLife Biochemistry and Chemical Biology The post-translational modification of histones by the small ubiquitin-like modifier (SUMO) protein has been associated with gene regulation, centromeric localization, and double-strand break repair in eukaryotes. Although sumoylation of histone H4 was specifically associated with gene repression, this could not be proven due to the challenge of site-specifically sumoylating H4 in cells. Biochemical crosstalk between SUMO and other histone modifications, such as H4 acetylation and H3 methylation, that are associated with active genes also remains unclear. We addressed these challenges in mechanistic studies using an H4 chemically modified at Lys12 by SUMO-3 (H4K12su) and incorporated into mononucleosomes and chromatinized plasmids for functional studies. Mononucleosome-based assays revealed that H4K12su inhibits transcription-activating H4 tail acetylation by the histone acetyltransferase p300, as well as transcription-associated H3K4 methylation by the extended catalytic module of the Set1/COMPASS (complex of proteins associated with Set1) histone methyltransferase complex. Activator- and p300-dependent in vitro transcription assays with chromatinized plasmids revealed that H4K12su inhibits both H4 tail acetylation and RNA polymerase II-mediated transcription. Finally, cell-based assays with a SUMO-H4 fusion that mimics H4 tail sumoylation confirmed the negative crosstalk between histone sumoylation and acetylation/methylation. Thus, our studies establish the key role for histone sumoylation in gene silencing and its negative biochemical crosstalk with active transcription-associated marks in human cells. eLife Sciences Publications, Ltd 2021-11-08 /pmc/articles/PMC8626089/ /pubmed/34747692 http://dx.doi.org/10.7554/eLife.67952 Text en © 2021, Leonen et al https://creativecommons.org/licenses/by/4.0/This article is distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use and redistribution provided that the original author and source are credited. |
spellingShingle | Biochemistry and Chemical Biology Leonen, Calvin Jon A Shimada, Miho Weller, Caroline E Nakadai, Tomoyoshi Hsu, Peter L Tyson, Elizabeth L Mishra, Arpit Shelton, Patrick MM Sadilek, Martin Hawkins, R David Zheng, Ning Roeder, Robert G Chatterjee, Champak Sumoylation of the human histone H4 tail inhibits p300-mediated transcription by RNA polymerase II in cellular extracts |
title | Sumoylation of the human histone H4 tail inhibits p300-mediated transcription by RNA polymerase II in cellular extracts |
title_full | Sumoylation of the human histone H4 tail inhibits p300-mediated transcription by RNA polymerase II in cellular extracts |
title_fullStr | Sumoylation of the human histone H4 tail inhibits p300-mediated transcription by RNA polymerase II in cellular extracts |
title_full_unstemmed | Sumoylation of the human histone H4 tail inhibits p300-mediated transcription by RNA polymerase II in cellular extracts |
title_short | Sumoylation of the human histone H4 tail inhibits p300-mediated transcription by RNA polymerase II in cellular extracts |
title_sort | sumoylation of the human histone h4 tail inhibits p300-mediated transcription by rna polymerase ii in cellular extracts |
topic | Biochemistry and Chemical Biology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8626089/ https://www.ncbi.nlm.nih.gov/pubmed/34747692 http://dx.doi.org/10.7554/eLife.67952 |
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