Graduate School of Frontier Biosciences, Osaka University, Suita, Osaka 565-0871, Japan.
J Cell Biol. 2009 Dec 14;187(6):781-90. doi: 10.1083/jcb.200904137.
Posttranslational histone modifications regulate both gene expression and genome integrity. Despite the dynamic nature of these modifications, appropriate real-time monitoring systems are lacking. In this study, we developed a method to visualize histone modifications in living somatic cells and preimplantation embryos by loading fluorescently labeled specific Fab antibody fragments. The technique was used to study histone H3 Ser10 (H3S10) phosphorylation, which occurs during chromosome condensation in mitosis mediated by the aurora B kinase. In aneuploid cancer cells that frequently missegregate chromosomes, H3S10 is phosphorylated just before the chromosomes condense, whereas aurora B already accumulates in nuclei during S phase. In contrast, in nontransformed cells, phosphorylated H3S10 foci appear for a few hours during interphase, and transient exposure to an aurora B-selective inhibitor during this period induces chromosome missegregation. These results suggest that, during interphase, moderate aurora B activity or H3S10 phosphorylation is required for accurate chromosome segregation. Visualizing histone modifications in living cells will facilitate future epigenetic and cell regulation studies.
翻译后组蛋白修饰调控基因表达和基因组完整性。尽管这些修饰具有动态性质，但缺乏合适的实时监测系统。在本研究中，我们开发了一种方法，通过加载荧光标记的特异性Fab抗体片段，在活体细胞和植入前胚胎中可视化组蛋白修饰。该技术用于研究组蛋白H3丝氨酸10（H3S10）磷酸化，其发生在有丝分裂过程中由极光B激酶介导的染色体凝聚期间。在经常发生染色体错分离的非整倍体癌细胞中，H3S10在染色体凝聚之前就被磷酸化，而极光B在S期就已在细胞核中积累。相比之下，在未转化细胞中，磷酸化的H3S10焦点在间期出现几个小时，在此期间短暂暴露于极光B选择性抑制剂会诱导染色体错分离。这些结果表明，在间期，适度的极光B活性或H3S10磷酸化对于准确的染色体分离是必需的。在活细胞中可视化组蛋白修饰将有助于未来的表观遗传学和细胞调控研究。
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