Mechanisms Underlying Enhancer Rnp Mediated Gene Regulation And Genome Organization

项目摘要 增强子在决定谱系特异性基因表达和细胞身份方面起关键作用， 富含非编码疾病突变和风险基因座。有一个迫切需要阐明完整的 增强子功能和故障的机制，以更好地了解疾病的病因学和开发新的， 增强子靶向治疗。最近发现活性增强子通常产生非编码增强子， RNA（eRNA）的研究为解释RNA介导的增强子活性提供了一个新的视角。 调控该提案旨在通过开发新的方法来建立一个独特的研究方向， 阐述eRNA在基因/基因组调控中的分子机制。我们的整体 假设eRNA组装特定的核糖核蛋白复合物（RNP）来组织高级有序结构， 三维核中的增强子结构来控制基因转录。我们有三个具体的 在这个提案中。我们计划使用我们已经开发的一种强大的RNA捕获方法， 鉴定人类细胞中eRNA：蛋白质相互作用组。对于第一个目标，我们计划集中在两个新的eRNA- 结合蛋白（eRBP），以表征其在基因转录和3D基因组相互作用中的功能， 哺乳动物细胞随后，我们有两个目的，研究这两个eRBP的机制，以充分 表征允许eRNA的生化基础：蛋白质相互作用及其可能参与 介导转录相关的亚核缩合物的形成。的预期成果 这一建议将为基因转录调控和RNA功能提供新的机制见解， 哺乳动物细胞随着增强子成为疾病发展的重要驱动力，这些机制性的见解 可能为治疗增强子驱动的疾病提供新的治疗策略。

PROJECT SUMMARY Enhancers play critical roles in determining lineage-specific gene expression and cellular identities, and are enriched in noncoding disease mutations and risk loci. There is an urgent need to elucidate the complete mechanisms of enhancer functions and malfunctions to better understand disease etiology and to develop new, enhancer-targeting therapies. The recent finding that active enhancers often generate noncoding enhancer RNAs (eRNAs) has provided a new perspective to interpret enhancer activity in the context of RNA-mediated regulation. This proposal aims to establish a unique research direction by developing new approaches to address the molecular mechanisms underlying eRNA functions in gene/genome regulation. Our overall hypothesis is that eRNAs assemble specific ribonucleoprotein complexes (RNPs) to organize high-order enhancer structures in the three-dimensional nucleus to control gene transcription. We have three specific aims in this proposal. We plan to use a robust RNA capture method that we have developed to systematically identify eRNA:protein interactomes in human cells. For the first aim, we plan to focus on two novel eRNA- binding proteins (eRBPs) to characterize their functions in gene transcription and 3D genome interaction in mammalian cells. Subsequently, we have two aims to study the mechanisms of each of the two eRBPs to fully characterize the biochemical basis permitting eRNA:protein interaction and their possible involvement in mediating the formation of transcriptionally-associated sub-nuclear condensates. The expected results from this proposal will provide novel mechanistic insights into gene transcriptional regulation and RNA functions in mammalian cells. As enhancers become important drivers in disease development, these mechanistic insights may offer new therapeutic strategies to treat enhancer-driven diseases.