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首页> 外文期刊>Molecular and Cellular Biology >CTCF Interacts with and Recruits the Largest Subunit of RNA Polymerase II to CTCF Target Sites Genome-Wide
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CTCF Interacts with and Recruits the Largest Subunit of RNA Polymerase II to CTCF Target Sites Genome-Wide

机译：CTCF与全基因组范围内的CTCF目标位点相互作用并招募到最大的RNA聚合酶II亚基

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CTCF is a transcription factor with highly versatile functions ranging from gene activation and repression to the regulation of insulator function and imprinting. Although many of these functions rely on CTCF-DNA interactions, it is an emerging realization that CTCF-dependent molecular processes involve CTCF interactions with other proteins. In this study, we report the association of a subpopulation of CTCF with the RNA polymerase II (Pol II) protein complex. We identified the largest subunit of Pol II (LS Pol II) as a protein significantly colocalizing with CTCF in the nucleus and specifically interacting with CTCF in vivo and in vitro. The role of CTCF as a link between DNA and LS Pol II has been reinforced by the observation that the association of LS Pol II with CTCF target sites in vivo depends on intact CTCF binding sequences. “Serial” chromatin immunoprecipitation (ChIP) analysis revealed that both CTCF and LS Pol II were present at the β-globin insulator in proliferating HD3 cells but not in differentiated globin synthesizing HD3 cells. Further, a single wild-type CTCF target site (N-Myc-CTCF), but not the mutant site deficient for CTCF binding, was sufficient to activate the transcription from the promoterless reporter gene in stably transfected cells. Finally, a ChIP-on-ChIP hybridization assay using microarrays of a library of CTCF target sites revealed that many intergenic CTCF target sequences interacted with both CTCF and LS Pol II. We discuss the possible implications of our observations with respect to plausible mechanisms of transcriptional regulation via a CTCF-mediated direct link of LS Pol II to the DNA.
机译：CTCF是一种转录因子，具有从基因激活和抑制到调控绝缘子功能和印迹的多种功能。尽管这些功能中的许多功能都依赖于CTCF-DNA相互作用，但人们逐渐认识到CTCF依赖性分子过程涉及CTCF与其他蛋白质的相互作用。在这项研究中，我们报告了CTCF亚群与RNA聚合酶II（Pol II）蛋白复合物的关联。我们确定Pol II的最大亚基（LS Pol II）是与CTCF在细胞核中显着共定位并在体内和体外与CTCF特异性相互作用的蛋白质。观察到LS Pol II与体内CTCF靶位的结合取决于完整的CTCF结合序列，从而加强了CTCF作为DNA与LS Pol II之间的联系的作用。 “串行”染色质免疫沉淀（ChIP）分析表明，CTCF和LS Pol II都存在于增殖的HD3细胞的β-珠蛋白绝缘子中，但不存在于合成珠蛋白的HD3细胞中。此外，单个野生型CTCF靶位点（N-Myc-CTCF）足以激活稳定转染的细胞中无启动子报告基因的转录，但不足以实现CTCF结合缺陷的突变位点。最后，使用CTCF目标位点文库的微阵列进行的ChIP-on-ChIP杂交分析显示，许多基因间CTCF目标序列均与CTCF和LS Pol II相互作用。我们讨论了有关通过CTCF介导的LS Pol II与DNA的直接连接的转录调控的合理机制的观察结果的可能含义。

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