Distribution of histone H4 modifications as revealed by a panel of specific monoclonal antibodies

Chromosome Research : an International Journal on the Molecular, Supramolecular and Evolutionary Aspects of Chromosome Biology
Yoko Hayashi-TakanakaHiroshi Kimura

Abstract

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 si...Continue Reading

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Citations

Apr 12, 2016·Scientific Reports·Jun-Ya KaimoriYoshitaka Isaka
Aug 19, 2016·Journal of Molecular Biology·Yuko SatoHiroshi Kimura
Oct 18, 2016·Journal of Cellular Biochemistry·Fernanda WisnieskiMarília Cardoso Smith
Aug 7, 2018·Epigenetics : Official Journal of the DNA Methylation Society·Lusy HandokoTakashi Umehara
Jan 18, 2018·Proceedings of the National Academy of Sciences of the United States of America·Christopher JennessHironori Funabiki
Nov 5, 2016·Nature Communications·Wei-Hao ShangTatsuo Fukagawa
Jun 25, 2020·Journal of Cell Science·Yoko Hayashi-TakanakaHiroshi Kimura
Dec 13, 2016·The Journal of Cell Biology·Tetsuya HoriTatsuo Fukagawa
Mar 23, 2017·Cell Death Discovery·Marina PartolinaAlexander Kagansky
Sep 16, 2020·Genes to Cells : Devoted to Molecular & Cellular Mechanisms·Erina SatomiMiki Hieda
Jan 21, 2021·Proceedings of the National Academy of Sciences of the United States of America·Yusuke FujiwaraMotomu Kanai
Feb 13, 2021·Life Science Alliance·Cheng-Han HoHitoshi Kurumizaka
Sep 4, 2021·Molecular Cell·Yasuhiro ArimuraHironori Funabiki

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Datasets Mentioned

BETA
SRX185820

Methods Mentioned

BETA
acetylation
acetylations
ELISA
ChIP
PCR
immunoprecipiation
ChIP-seq
Electrophoresis
flow cytometry

Software Mentioned

genome
Cufflinks
browser
Tophat
Bowtie

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