Live-cell imaging reveals the dynamics of PRC2 and recruitment to chromatin by SUZ12-associated subunits.
Live-cell imaging reveals the dynamics of PRC2 and recruitment to chromatin by SUZ12-associated subunits.
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DOI：
10.1101/gad.311936.118
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发表时间：
2018-06-01
影响因子：
10.5
通讯作者：
Cech TR
中科院分区：
文献类型：
--
作者：
Youmans DT;Schmidt JC;Cech TR
Youmans等人在活细胞中进行了单粒子成像，并显示了PRC 2染色质修饰剂的核分布和动力学。他们的发现量化了PRC 2与人类癌细胞中染色质的结合，并分离了H3 K27 me 3组蛋白标记和各种PRC 2亚基对PRC 2向染色质募集的贡献。多梳抑制复合物2（PRC 2）是一种促进表观遗传基因沉默的组蛋白甲基转移酶，但其与染色质相互作用的动力学在很大程度上是未知的。在这里，我们定量测量了PRC 2与人类癌细胞中染色质的结合。将HaloTag编辑到内源性EZH 2和SUZ 12基因座中的基因组编辑和单粒子追踪显示，PRC 2的约80%快速扩散通过细胞核，而约20%是染色质结合的。用EED-H3 K27 me 3相互作用的小分子抑制剂进行短期处理对PRC 2的染色质停留时间没有立即影响。相反，SUZ 12的功能分离突变体仍然形成核心PRC 2复合物，但不能结合辅助蛋白，揭示了AEBP 2和PCL同源蛋白对染色质结合的主要贡献。因此，我们量化了这种染色质修饰复合物在活细胞中的动力学，并分离了H3 K27 me 3组蛋白标记和各种PRC 2亚基对PRC 2向染色质募集的贡献。
Here, Youmans et al. performed single-particle imaging in live cells and show the nuclear distribution and dynamics of the PRC2 chromatin modifier. Their findings quantify the binding of PRC2 to chromatin in human cancer cells and separate the contributions of H3K27me3 histone marks and various PRC2 subunits to recruitment of PRC2 to chromatin. Polycomb-repressive complex 2 (PRC2) is a histone methyltransferase that promotes epigenetic gene silencing, but the dynamics of its interactions with chromatin are largely unknown. Here we quantitatively measured the binding of PRC2 to chromatin in human cancer cells. Genome editing of a HaloTag into the endogenous EZH2 and SUZ12 loci and single-particle tracking revealed that ∼80% of PRC2 rapidly diffuses through the nucleus, while ∼20% is chromatin-bound. Short-term treatment with a small molecule inhibitor of the EED–H3K27me3 interaction had no immediate effect on the chromatin residence time of PRC2. In contrast, separation-of-function mutants of SUZ12, which still form the core PRC2 complex but cannot bind accessory proteins, revealed a major contribution of AEBP2 and PCL homolog proteins to chromatin binding. We therefore quantified the dynamics of this chromatin-modifying complex in living cells and separated the contributions of H3K27me3 histone marks and various PRC2 subunits to recruitment of PRC2 to chromatin.
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通讯作者：
Di Croce, Luciano