The organization of histone H3 modifications as revealed by a panel of specific monoclonal antibodies
| Autor: | Yoko Hayashi-Takanaka, Nanako Takizawa, Naohito Nozaki, Hiroshi Kimura, Yuji Goto |
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| Rok vydání: | 2008 |
| Předmět: |
Antibody Specificity
Chromatin Immunoprecipitation Physiology Immunoblotting Fluorescent Antibody Technique Biology Antibodies Monoclonal/*immunology Methylation Histones Cell Aging/immunology Histone H3 Mice Histone H1 Nucleosomes/immunology/metabolism Fibroblasts/cytology/immunology/metabolism Histone H2A Histone methylation Histone code Animals Humans Molecular Biology Cellular Senescence Epigenomics Lysine Antibodies Monoclonal Acetylation Cell Biology General Medicine Histones/*chemistry/immunology/*metabolism Fibroblasts Molecular biology Nucleosomes Histone methyltransferase Chromatin immunoprecipitation Lysine/metabolism |
| Zdroj: | Cell structure and function. 33(1) |
| ISSN: | 1347-3700 |
| Popis: | Histone modifications play critical roles in the epigenetic regulation of gene expression and in the maintenance of genome integrity. Acetylation and methylation of histone H3 are particularly important in gene activation and silencing. We generated and characterized a panel of mouse monoclonal antibodies that specifically recognize different modifications on K4, K9, and K27 residues on histone H3. By using these antibodies for chromatin immunoprecipitation and immunoblotting, we analyzed the relationship between different modifications in nearby nucleosomes in human cells. Within a few nucleosome neighbors, trimethyl-K4 was associated with acetyl-K27, rather than with dimethyl-K4 and acetyl-K9, consistent with their co-localization on active promoters. Furthermore, simultaneous immunofluorescence using directly-labeled antibodies revealed that di- and tri-methylation on K4 was diminished during replicative senescence. These highly-reliable and fully-characterized monoclonal antibodies may facilitate future epigenomic studies on healthy and diseased cells. |
| Databáze: | OpenAIRE |
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