CTCF Binding Polarity Determines Chromatin Looping
CTCF Binding Polarity Determines Chromatin Looping
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DOI：
10.1016/j.molcel.2015.09.023
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发表时间：
2015-11-19
期刊：
影响因子：
16
通讯作者：
de laat, Wouter
中科院分区：
文献类型：
--
作者：
de Wit, Elzo;Vos, Erica S. M.;de laat, Wouter
CCCTC结合因子(CTCF)是一种参与染色质三维组织的结构蛋白。在这项研究中，我们用高分辨率的4C-seq分析了大量CTCF结合位点的3D基因组接触图谱。正如最近报道的那样，我们的数据还表明，染色质环优先在CTCF结合位点之间以收敛的方式形成。为了直接测试这一点，我们使用CRISPR/Cas9基因组编辑删除了三个座位上的核心CTCF结合位点，包括Sox2超级增强子中的CTCF位点。在所有的病例中，CTCF和粘附素的重新募集都丢失了，与远端、汇聚的CTCF位点的染色质环被破坏或不稳定。重新插入相反方向的CTCF识别序列恢复了CTCF和粘附素的招募，但没有重新建立染色质环。我们得出结论，CTCF结合极性在高阶染色质结构的形成中起着功能作用。
CCCTC-binding factor (CTCF) is an architectural protein involved in the three-dimensional (3D) organization of chromatin. In this study, we assayed the 3D genomic contact profiles of a large number of CTCF binding sites with high-resolution 4C-seq. As recently reported, our data also suggest that chromatin loops preferentially form between CTCF binding sites oriented in a convergent manner. To directly test this, we used CRISPR/Cas9 genome editing to delete core CTCF binding sites in three loci, including the CTCF site in the Sox2 super-enhancer. In all instances, CTCF and cohesin recruitment were lost, and chromatin loops with distal, convergent CTCF sites were disrupted or destabilized. Re-insertion of oppositely oriented CTCF recognition sequences restored CTCF and cohesin recruitment, but did not re-establish chromatin loops. We conclude that CTCF binding polarity plays a functional role in the formation of higher-order chromatin structure.