Characterization of a novel family of human transcription factors that bind at +240 downstream of the transcription start site.

我们的研究集中在一个由ZFX、ZFY和ZNF 711组成的人类C2 H2锌指蛋白家族 在所有人类细胞类型中都有表达。在我们的初步研究中，我们发现ZFX的缺失 家族成员对细胞增殖具有严重的有害后果。我们进行了ChIP- 在几种不同的人细胞系中进行ZFX、ZFY和ZNF 711的seq测定。有趣的是，我们发现， 这些TF在相同的CpG岛启动子区域具有相同的结合模式， 在转录起始位点下游+240bp处结合。尽管它们的蛋白质结构表明 ZFX、ZFY和ZNF 711是转录调节因子，它们影响转录的机制 转录尚未阐明。我们的初步结果表明，ZXF家族成员 是人类转录组的重要调节因子。未能适当监管 转录组可导致多种人类疾病。因此，彻底描述这一点 TF家族至关重要。两个家族成员（ZFX和ZFY）基本相同 在X或Y染色体上编码的蛋白质，而ZNF 711与ZFX具有67%的总体相似性 锌指结构域的相似性为87%。因为ZFX和ZFY基本上是相同的蛋白质，我们将 集中于使用雌性细胞系（其缺乏ZFY）比较ZFX和ZNF 711。我们建议 描述这些因子通过三管齐下的方法调节转录的机制。在 目的1，我们将进行ZFX和ZNF 711结合的详细研究，以确定这些结合是如何发生的。 转录因子被募集到CpG岛启动子。在目标2中，我们将确定关键的监管 ZFX和ZNF 711的结构域和相互作用蛋白。在目标3中，我们将通过以下方式描述该机制： ZFX和ZNF 711介导转录调控。重要的是，每个目标都可以实现 独立于其他人，并可以立即开始开始。通过目标1-3的结合，我们将 使用表观基因组学、蛋白质组学和转录组学方法彻底表征ZFX家族的功能。 方法。完成我们提出的研究将提供新的见解转录 调控和染色质结构在人类CpG岛启动子。

Our study focuses on a family of human C2H2 zinc finger proteins composed of ZFX, ZFY, and ZNF711 which are expressed in all human cell types. In our preliminary studies, we show that deletion of ZFX family members has severe detrimental consequences on cell proliferation. We have performed ChIP- seq assays for ZFX, ZFY, and ZNF711 in several different human cell lines. Interestingly, we found that these TFs have identical binding patterns at the same CpG island promoter regions, with an average peak of binding at +240bp downstream of the transcription start site. Although their protein structure suggests that ZFX, ZFY, and ZNF711 are transcriptional regulators, the mechanisms by which they influence transcription have not yet been elucidated. Our preliminary results suggest that the ZXF family members are important regulators of the human transcriptome. A failure to properly regulate the transcriptome can lead to many types of human disease. Therefore, a thorough characterization of this family of TFs is of critical importance. Two of the family members (ZFX and ZFY) are essentially identical proteins encoded on either the X or Y chromosome, whereas ZNF711 has 67% overall similarity with ZFX and 87% similarity in the zinc finger domain. Because ZFX and ZFY are basically identical proteins, we will focus on a comparison of ZFX and ZNF711 using a female cell line (which lacks ZFY). We propose to characterize the mechanisms by which these factors regulate transcription using a 3-pronged approach. In Aim 1, we will perform detailed investigations of ZFX and ZNF711 binding to determine how these transcription factors are recruited to CpG island promoters. In Aim 2, we will identify critical regulatory domains and interacting proteins of ZFX and ZNF711. In Aim 3, we will characterize the mechanism by which ZFX and ZNF711 mediate transcriptional regulation. Importantly, each of the Aims can be performed independently of the others and can begin immediately. Through the combination of Aims 1-3, we will thoroughly characterize the function of the ZFX family using epigenomic, proteomic, and transcriptomic approaches. Completion of our proposed studies will provide new insights into transcriptional regulation and chromatin structure at human CpG island promoters.