Mechanisms Underlying Enhancer Rnp Mediated Gene Regulation And Genome Organization

项目摘要 增强子在确定谱系特异性基因表达和细胞特性方面起着关键作用，并且 富含非编码疾病突变和危险基因。迫切需要澄清完整的 增强子功能和功能障碍的机制，以更好地了解疾病的病因，并开发新的， 增强型靶向疗法。最近的研究发现，活性增强子经常产生非编码增强子 RNAs(ERNAs)为解释RNA介导的增强子活性提供了新的视角 监管。这项建议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/基因组调控中潜在作用的分子机制。我们的整体 假设eRNAs组装特定的核糖核蛋白复合体(RNPs)来组织高阶 三维核中控制基因转录的增强子结构。我们有三个具体的 在这项提案中的目标。我们计划使用我们开发的一种强大的RNA捕获方法来系统地 鉴定Erna：人类细胞中的蛋白质相互作用。对于第一个目标，我们计划集中在两部小说厄娜- 结合蛋白(ERBPs)研究其在基因转录和3D基因组相互作用中的功能 哺乳动物细胞。随后，我们有两个目的来充分研究这两个eRBP的作用机制 表征允许ERNA的生化基础：蛋白质相互作用及其可能参与的 调节转录相关的亚核凝聚体的形成。预期结果来自 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。随着增强剂成为疾病发展的重要驱动力，这些机械性的见解 可能为治疗增强剂驱动的疾病提供新的治疗策略。

PROJECT ABSTRACT 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.