caa a22 4500 606190627 CHVBK 20210128100856.0 cr unu---uuuuu 210128e20151201xx s 000 0 eng 10.1007/s10577-015-9486-4 doi (NATIONALLICENCE)springer-10.1007/s10577-015-9486-4 Distribution of histone H4 modifications as revealed by a panel of specific monoclonal antibodies [Elektronische Daten] [Yoko Hayashi-Takanaka, Kazumitsu Maehara, Akihito Harada, Takashi Umehara, Shigeyuki Yokoyama, Chikashi Obuse, Yasuyuki Ohkawa, Naohito Nozaki, Hiroshi Kimura] 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. The Author(s), 2015 Chromatin nationallicence Epigenetics nationallicence Histone modification nationallicence Monoclonal antibody nationallicence HAT : Histone acetyltransferase nationallicence mAbs : Monoclonal antibodies nationallicence PBS : Phosphate-buffered saline nationallicence EdU : Ethynyl-deoxyuridine nationallicence TSSs : Transcription start sites nationallicence Hayashi-Takanaka Yoko Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, 4259 Nagatsuta, Midori-ku, 226-8501, Yokohama, Japan aut Maehara Kazumitsu Department of Advanced Medical Initiatives, Faculty of Medicine, Kyushu University, 812-8582, Fukuoka, Japan aut Harada Akihito Department of Advanced Medical Initiatives, Faculty of Medicine, Kyushu University, 812-8582, Fukuoka, Japan aut Umehara Takashi RIKEN Center for Life Science Technologies, 230-0045, Yokohama, Japan aut Yokoyama Shigeyuki RIKEN Structural Biology Laboratory, 230-0045, Yokohama, Japan aut Obuse Chikashi School of Life Sciences, Hokkaido University, 001-0021, Sapporo, Japan aut Ohkawa Yasuyuki CREST, JST, 332-0012, Kawaguchi, Saitama, Japan aut Nozaki Naohito MAB Institute Inc, 001-0021, Sapporo, Japan aut Kimura Hiroshi Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, 4259 Nagatsuta, Midori-ku, 226-8501, Yokohama, Japan aut Chromosome Research Springer Netherlands 23/4(2015-12-01), 753-766 0967-3849 23:4<753 2015 23 10577 https://doi.org/10.1007/s10577-015-9486-4 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s10577-015-9486-4 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Hayashi-Takanaka Yoko Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, 4259 Nagatsuta, Midori-ku, 226-8501, Yokohama, Japan aut NATIONALLICENCE 700 1- Maehara Kazumitsu Department of Advanced Medical Initiatives, Faculty of Medicine, Kyushu University, 812-8582, Fukuoka, Japan aut NATIONALLICENCE 700 1- Harada Akihito Department of Advanced Medical Initiatives, Faculty of Medicine, Kyushu University, 812-8582, Fukuoka, Japan aut NATIONALLICENCE 700 1- Umehara Takashi RIKEN Center for Life Science Technologies, 230-0045, Yokohama, Japan aut NATIONALLICENCE 700 1- Yokoyama Shigeyuki RIKEN Structural Biology Laboratory, 230-0045, Yokohama, Japan aut NATIONALLICENCE 700 1- Obuse Chikashi School of Life Sciences, Hokkaido University, 001-0021, Sapporo, Japan aut NATIONALLICENCE 700 1- Ohkawa Yasuyuki CREST, JST, 332-0012, Kawaguchi, Saitama, Japan aut NATIONALLICENCE 700 1- Nozaki Naohito MAB Institute Inc, 001-0021, Sapporo, Japan aut NATIONALLICENCE 700 1- Kimura Hiroshi Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, 4259 Nagatsuta, Midori-ku, 226-8501, Yokohama, Japan aut NATIONALLICENCE 773 0- Chromosome Research Springer Netherlands 23/4(2015-12-01), 753-766 0967-3849 23:4<753 2015 23 10577