The Organization of Histone H3 Modifications as Revealed by a Panel of Specific Monoclonal Antibodies
The Organization of Histone H3 Modifications as Revealed by a Panel of Specific Monoclonal Antibodies
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DOI：
10.1247/csf.07035
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发表时间：
2008-01-01
影响因子：
1.5
通讯作者：
Nozaki, Naohito
中科院分区：
文献类型：
--
作者：
Kimura, Hiroshi;Hayashi-Takanaka, Yoko;Nozaki, Naohito
组蛋白修饰在基因表达的表观遗传调控和基因组完整性的维持中起着至关重要的作用。组蛋白H3的乙酰化和甲基化在基因激活和沉默中特别重要。我们生成并表征了一组小鼠单克隆抗体，其特异性识别组蛋白H3上K4、K9和K27残基上的不同修饰。通过使用这些抗体进行染色质免疫沉淀和免疫印迹，我们分析了人类细胞中邻近核小体中不同修饰之间的关系。在一些核小体的邻居，三甲基-K4与乙酰基-K27，而不是与二甲基-K4和乙酰基-K9，与他们的共定位活性启动子一致。此外，同时使用直接标记的抗体的免疫荧光显示，K4的二甲基化和三甲基化在复制衰老过程中减少。这些高度可靠和完全表征的单克隆抗体可能有助于未来对健康和患病细胞的表观基因组研究。
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.