| RRC ID |
37917
|
| 著者 |
Hayashi-Takanaka Y, Maehara K, Harada A, Umehara T, Yokoyama S, Obuse C, Ohkawa Y, Nozaki N, Kimura H.
|
| タイトル |
Distribution of histone H4 modifications as revealed by a panel of specific monoclonal antibodies.
|
| ジャーナル |
Chromosome Res
|
| 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 sites. These extensively characterized and highly specific antibodies will be useful for future epigenetics and epigenome studies.
|
| 巻・号 |
23(4)
|
| ページ |
753-66
|
| 公開日 |
2015-12-1
|
| DOI |
10.1007/s10577-015-9486-4
|
| PII |
10.1007/s10577-015-9486-4
|
| PMID |
26343042
|
| PMC |
PMC4666908
|
| MeSH |
Acetylation
Animals
Antibodies, Monoclonal* / metabolism
Blotting, Western
Cell Line, Transformed
Chromatin / genetics
Chromatin / metabolism
Chromatin Immunoprecipitation
Epigenesis, Genetic
HeLa Cells
High-Throughput Nucleotide Sequencing
Histones / metabolism*
Humans
Immunoglobulin G
Immunohistochemistry
Methylation
Mice
Protein Processing, Post-Translational*
|
| IF |
3.413
|
| 引用数 |
20
|
|
WOS 分野
|
GENETICS & HEREDITY
BIOCHEMISTRY & MOLECULAR BIOLOGY
|
| リソース情報 |
| ヒト・動物細胞 |
ZHBTc4(AES0136) |
