Phosphorylation of H3S10 Blocks the Access of H3K9 by Specific Antibodies and Histone Methyltransferase IMPLICATION IN REGULATING CHROMATIN DYNAMICS AND EPIGENETIC INHERITANCE DURING MITOSIS
Phosphorylation of H3S10 Blocks the Access of H3K9 by Specific Antibodies and Histone Methyltransferase IMPLICATION IN REGULATING CHROMATIN DYNAMICS AND EPIGENETIC INHERITANCE DURING MITOSIS
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DOI：
10.1074/jbc.m803312200
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发表时间：
2008-11-28
影响因子：
4.8
通讯作者：
Dai, Wei
中科院分区：
文献类型：
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作者：
Duan, Qing;Chen, Haobin;Dai, Wei
组蛋白的翻译后修饰在调节基因组结构和完整性方面起着关键作用。我们集中在组蛋白H3赖氨酸9（H3K9）和H3S10在细胞周期中的共价修饰之间的调节关系。免疫荧光显微镜显示，在HeLa，A549和HCT 116细胞中的H3S10磷酸化是高的前期，前中期和中期，而H3K9单甲基化（H3K9me1）和二甲基化（H3K9me2），但不是H3K9三甲基化（H3K9me3），显着抑制。当H3S10磷酸化在后期开始减少时，H3K9me1和H3K9me2信号重新出现。Western印迹分析证实，在含有SDS的缓冲液中提取的有丝分裂组蛋白几乎没有H3K9me1和H3K9me2信号，但有丰富的H3K9me3信号。然而，当在没有SDS的相同缓冲液中提取有丝分裂组蛋白时，间期细胞和有丝分裂细胞之间的H3K9me1和H3K9me2信号的差异消失。通过荧光显微镜和蛋白质印迹检测，通过用磷酸化酶预处理去除H3S10磷酸化，暴露了有丝分裂H3K9me1和H3K9me2信号。此外，H3S10磷酸化在体外完全阻断H3K9的甲基化，但不阻断相同残基的去甲基化。鉴于几个保守的基序组成的一个赖氨酸残基，紧接着一个丝氨酸残基存在于组蛋白尾部，我们的研究揭示了一个潜在的新机制，磷酸化不仅调节细胞因子的甲基化赖氨酸的选择性访问，但也有助于保持甲基化模式和细胞分裂过程中的表观遗传程序。
Post-translational modifications of histones play a critical role in regulating genome structures and integrity. We have focused on the regulatory relationship between covalent modifications of histone H3 lysine 9 (H3K9) and H3S10 during the cell cycle. Immunofluorescence microscopy revealed that H3S10 phosphorylation in HeLa, A549, and HCT116 cells was high during prophase, prometaphase, and metaphase, whereas H3K9 monomethylation (H3K9me1) and dimethylation (H3K9me2), but not H3K9 trimethylation (H3K9me3), were significantly suppressed. When H3S10 phosphorylation started to diminish during anaphase, H3K9me1 and H3K9me2 signals reemerged. Western blot analyses confirmed that mitotic histones, extracted in an SDS-containing buffer, had little H3K9me1 and H3K9me2 signals but abundant H3K9me3 signals. However, when mitotic histones were extracted in the same buffer without SDS, the difference in H3K9me1 and H3K9me2 signals between interphase and mitotic cells disappeared. Removal of H3S10 phosphorylation by pretreatment with lambda-phosphatase unmasked mitotic H3K9me1 and H3K9me2 signals detected by both fluorescence microscopy and Western blotting. Further, H3S10 phosphorylation completely blocked methylation of H3K9 but not demethylation of the same residue in vitro. Given that several conserved motifs consisting of a Lys residue immediately followed by a Ser residue are present in histone tails, our studies reveal a potential new mechanism by which phosphorylation not only regulates selective access of methylated lysines by cellular factors but also serves to preserve methylation patterns and epigenetic programs during cell division.