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Distribution of histone H4 modifications as revealed by a panel of specific monoclonal antibodies
Post-translational histone modifications play a critical role in genome functions such as epigenetic gene regulation and genome maintenance. The tail of the histone H4 N-terminus contains several amino acids that can be acetylated and methylated. Some of these modifications are known to undergo dras...
Autores principales: | , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Springer Netherlands
2015
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4666908/ https://www.ncbi.nlm.nih.gov/pubmed/26343042 http://dx.doi.org/10.1007/s10577-015-9486-4 |
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author | Hayashi-Takanaka, Yoko Maehara, Kazumitsu Harada, Akihito Umehara, Takashi Yokoyama, Shigeyuki Obuse, Chikashi Ohkawa, Yasuyuki Nozaki, Naohito Kimura, Hiroshi |
author_facet | Hayashi-Takanaka, Yoko Maehara, Kazumitsu Harada, Akihito Umehara, Takashi Yokoyama, Shigeyuki Obuse, Chikashi Ohkawa, Yasuyuki Nozaki, Naohito Kimura, Hiroshi |
author_sort | Hayashi-Takanaka, Yoko |
collection | PubMed |
description | Post-translational histone modifications play a critical role in genome functions such as epigenetic gene regulation and genome maintenance. The tail of the histone H4 N-terminus contains several amino acids that can be acetylated and methylated. Some of these modifications are known to undergo drastic changes during the cell cycle. In this study, we generated a panel of mouse monoclonal antibodies against histone H4 modifications, including acetylation at K5, K8, K12, and K16, and different levels of methylation at K20. Their specificity was evaluated by ELISA and immunoblotting using synthetic peptide and recombinant proteins that harbor specific modifications or amino acid substitutions. Immunofluorescence confirmed the characteristic distributions of target modifications. An H4K5 acetylation (H4K5ac)-specific antibody CMA405 reacted with K5ac only when the neighboring K8 was unacetylated. This unique feature allowed us to detect newly assembled H4, which is diacetylated at K5 and K12, and distinguish it from hyperacetylated H4, where K5 and K8 are both acetylated. Chromatin immunoprecipiation combined with deep sequencing (ChIP-seq) revealed that acetylation of both H4K8 and H4K16 were enriched around transcription start sites. These extensively characterized and highly specific antibodies will be useful for future epigenetics and epigenome studies. |
format | Online Article Text |
id | pubmed-4666908 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2015 |
publisher | Springer Netherlands |
record_format | MEDLINE/PubMed |
spelling | pubmed-46669082015-12-09 Distribution of histone H4 modifications as revealed by a panel of specific monoclonal antibodies Hayashi-Takanaka, Yoko Maehara, Kazumitsu Harada, Akihito Umehara, Takashi Yokoyama, Shigeyuki Obuse, Chikashi Ohkawa, Yasuyuki Nozaki, Naohito Kimura, Hiroshi Chromosome Res Article Post-translational histone modifications play a critical role in genome functions such as epigenetic gene regulation and genome maintenance. The tail of the histone H4 N-terminus contains several amino acids that can be acetylated and methylated. Some of these modifications are known to undergo drastic changes during the cell cycle. In this study, we generated a panel of mouse monoclonal antibodies against histone H4 modifications, including acetylation at K5, K8, K12, and K16, and different levels of methylation at K20. Their specificity was evaluated by ELISA and immunoblotting using synthetic peptide and recombinant proteins that harbor specific modifications or amino acid substitutions. Immunofluorescence confirmed the characteristic distributions of target modifications. An H4K5 acetylation (H4K5ac)-specific antibody CMA405 reacted with K5ac only when the neighboring K8 was unacetylated. This unique feature allowed us to detect newly assembled H4, which is diacetylated at K5 and K12, and distinguish it from hyperacetylated H4, where K5 and K8 are both acetylated. Chromatin immunoprecipiation combined with deep sequencing (ChIP-seq) revealed that acetylation of both H4K8 and H4K16 were enriched around transcription start sites. These extensively characterized and highly specific antibodies will be useful for future epigenetics and epigenome studies. Springer Netherlands 2015-09-05 2015 /pmc/articles/PMC4666908/ /pubmed/26343042 http://dx.doi.org/10.1007/s10577-015-9486-4 Text en © The Author(s) 2015 Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. |
spellingShingle | Article Hayashi-Takanaka, Yoko Maehara, Kazumitsu Harada, Akihito Umehara, Takashi Yokoyama, Shigeyuki Obuse, Chikashi Ohkawa, Yasuyuki Nozaki, Naohito Kimura, Hiroshi Distribution of histone H4 modifications as revealed by a panel of specific monoclonal antibodies |
title | Distribution of histone H4 modifications as revealed by a panel of specific monoclonal antibodies |
title_full | Distribution of histone H4 modifications as revealed by a panel of specific monoclonal antibodies |
title_fullStr | Distribution of histone H4 modifications as revealed by a panel of specific monoclonal antibodies |
title_full_unstemmed | Distribution of histone H4 modifications as revealed by a panel of specific monoclonal antibodies |
title_short | Distribution of histone H4 modifications as revealed by a panel of specific monoclonal antibodies |
title_sort | distribution of histone h4 modifications as revealed by a panel of specific monoclonal antibodies |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4666908/ https://www.ncbi.nlm.nih.gov/pubmed/26343042 http://dx.doi.org/10.1007/s10577-015-9486-4 |
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