Visualization of the dynamic interaction between nucleosomal histone H3K9 tri-methylation and HP1α chromodomain in living cells
Visualization of the dynamic interaction between nucleosomal histone H3K9 tri-methylation and HP1α chromodomain in living cells
复制标题
活细胞中核小体组蛋白 H3K9 三甲基化与 HP1α 染色结构域之间动态相互作用的可视化
DOI：
10.1016/j.chembiol.2022.05.006
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发表时间：
2022
影响因子：
8.6
通讯作者：
Yoshida Minoru
中科院分区：
文献类型：
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作者：
Sasaki Kazuki;Suzuki Michihiro;Sonoda Takeshi;Schneider-Poetsch Tilman;Ito Akihiro;Takagi Motoki;Fujishiro Shinya;Sohtome Yoshihiro;Dodo Kosuke;Umehara Takashi;Aburatani Hiroyuki;Shin-ya Kazuo;Nakao Yoichi;Sodeoka Mikiko;Yoshida Minoru
组蛋白赖氨酸甲基化是控制基因表达的表观遗传标记。特别地，H3K9me3通过调节染色质结构而有助于转录抑制。成功的有丝分裂进程需要正确的染色质结构变化的时机，包括表观遗传标记。然而，活细胞中组蛋白修饰的时空信息仍然有限。在这项研究中，我们创建了一个基于FRET的探针，用于基于HP1α chromodomain（HP1αCD）的活细胞成像，该探针与H3K9me3结合。将探针掺入染色质中，用组蛋白甲基转移酶抑制剂处理后发射率降低，表明它成功地追踪了H3K9me3的动态变化。进入有丝分裂后，探针的发射率短暂增加，同时H3K9me3也随之增加，然后呈现逐步下降，可能是由于H3S10磷酸化和H3K9me3去甲基化导致HP 1 αCD结合丧失所致。该探针可用于检测与HP 1 α相关的染色质结构的动态变化。
Histone lysine methylation is an epigenetic mark that can control gene expression. In particular, H3K9me3 contributes to transcriptional repression by regulating chromatin structure. Successful mitotic progression requires correct timing of chromatin structure changes, including epigenetic marks. However, spatiotemporal information on histone modifications in living cells remains limited. In this study, we created an FRET-based probe for live-cell imaging based on the HP1α chromodomain (HP1αCD), which binds to H3K9me3. The probe was incorporated into chromatin and the emission ratio decreased after treatment with histone methyltransferase inhibitors, indicating that it successfully traced dynamic changes in H3K9me3. Upon entry into mitosis, the probe's emission ratio transiently increased with a concomitant increase in H3K9me3, then exhibited a stepwise decrease, probably due to loss of HP1αCD binding caused by phosphorylation of H3S10 and demethylation of H3K9me3. This probe will be a useful tool for detecting dynamic changes in chromatin structure associated with HP1α.