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

结合在转录起始位点下游 240 处的新型人类转录因子家族的表征。

• 批准号: 10589127
• 负责人: PEGGY J Farnham
• 金额: $ 33万
• 依托单位: UNIVERSITY OF SOUTHERN CALIFORNIA
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-04-01 至 2024-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10589127
• 关键词: Affect; Binding; Binding Sites; Biological Assay; C2H2 Zinc Finger; CRISPR/Cas technology; Cell Line; Cell Proliferation; Cells; ChIP-seq; Chromatin; Chromatin Structure; CpG Islands; DNA Binding; DNA Binding Domain; DNA Methylation; Disease; Elements; Failure; Family; Family member; Female; Genes; Genetic Transcription; Human; Human Cell Line; Investigation; Location; Maps; Mediating; Nuclear Extract; Nucleosomes; Pattern; Positioning Attribute; Promoter Regions; Proteins; Proteomics; Qualifying; RNA; Reporter; Role; Series; Techniques; Testing; Transcription Elongation; Transcription Initiation; Transcription Initiation Site; Transcriptional Regulation; Y Chromosome; ZFY protein; Zinc Fingers; cell type; epigenomics; genome-wide analysis; histone modification; human disease; insight; novel; novel therapeutics; promoter; protein protein interaction; protein structure; recruit; transcription factor; transcriptome; transcriptomics

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.

我们的研究重点是由 ZFX、ZFY 和 ZNF711 组成的人类 C2H2 锌指蛋白家族 它们在所有人类细胞类型中表达。在我们的初步研究中，我们表明删除 ZFX 家庭成员对细胞增殖有严重的不利影响。我们已经进行了 ChIP- 在几种不同的人类细胞系中对 ZFX、ZFY 和 ZNF711 进行 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，平均峰值 结合在转录起始位点下游+240bp处。尽管它们的蛋白质结构表明 ZFX、ZFY 和 ZNF711 是转录调节因子，它们影响的机制 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。未能正确监管 转录组可以导致多种类型的人类疾病。因此，对这个问题进行全面的表征 TF家族至关重要。两个家族成员（ZFX 和 ZFY）本质上是相同的 X 或 Y 染色体上编码的蛋白质，而 ZNF711 与 ZFX 总体相似度为 67% 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，所以我们将 重点是使用雌性细胞系（缺乏 ZFY）对 ZFX 和 ZNF711 进行比较。我们建议 使用三管齐下的方法描述这些因子调节转录的机制。在 目标 1，我们将对 ZFX 和 ZNF711 结合进行详细研究，以确定这些结合如何 转录因子被招募到 CpG 岛启动子上。在目标 2 中，我们将确定关键监管 ZFX 和 ZNF711 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： 其中 ZFX 和 ZNF711 介导转录调节。重要的是，每个目标都可以实现 独立于其他人并且可以立即开始。通过目标 1-3 的结合，我们将 使用表观基因组、蛋白质组和转录组彻底表征 ZFX 家族的功能 接近。完成我们提出的研究将为转录提供新的见解 人类 CpG 岛启动子的调控和染色质结构。