Insulators: Determinants of chromosome topology and regulatory interactions

绝缘体：染色体拓扑和调控相互作用的决定因素

• 批准号: 9349572
• 负责人: JAMES B JAYNES
• 金额: $ 31.2万
• 依托单位: THOMAS JEFFERSON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-09-08 至 2020-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9349572
• 关键词: 3-Dimensional; Address; Affect; Aging-Related Process; Animals; Architecture; Binding Proteins; Biochemical; Biological Assay; Cell Nucleus; Chromatin; Chromosomes; Code; Complex; DNA; DNA Sequence; Development; Diagnosis; Diagnostic; Disease; Dissection; Drosophila genus; Elements; Embryonic Development; Enhancers; Eukaryota; Gel; Gene Expression; Gene Expression Regulation; Genes; Genetic Transcription; Genome; Human; Image; Inborn Genetic Diseases; Inherited; Intervention; Laboratories; Length; Mammals; Mediating; Molecular; Molecular Conformation; Neighborhoods; Nuclear; Nuclear Extract; Organism; Pattern; Play; Polycomb; Prevention; Process; Property; Protein Array; Proteins; Recruitment Activity; Regulator Genes; Research; Role; Specificity; Testing; Transgenes; Transgenic Organisms; base; cell type; combat; combinatorial; design; direct application; effective intervention; genome-wide; human disease; in vivo; insight; novel; novel strategies; promoter; protein complex; tool; tumor progression; tumorigenesis

Changes in gene transcription are important in the progression of cancer, in most other human diseases, and in the aging process, as well as in the development of multicellular organisms at all stages. In recent years it has become clear that chromosome topology plays a vital role in gene regulation, as well as other nuclear processes. The major determinant of this topology involves regulatory DNA known as insulators. Tools available in Drosophila make it possible to study mechanisms of insulator action and interaction in detail, in a true in vivo context. This proposal is to study mechanisms of chromatin-based gene regulation involving insulators, including those recently discovered in the well-characterized Drosophila gene even skipped. These studies will address basic questions of how regulatory DNA that mediates chromosome topology carries out its function in 3 dimensions inside the nucleus. They will show how chromosome architecture interfaces with alternative transcriptional states by integrating the topology of insulator pairing with that of enhancer-promoter interactions. A unique contribution of these studies will be to identify an insulator pairing code that can be used to predict and design chromosome architecture and the consequence effects on gene expression. In mammals, insulators are known to be involved in cell-type specific gene regulation and in key developmental decisions, as well as in oncogenesis and inherited human disorders. Therefore, these studies will have direct applications to research on human diseases. The Specific Aims are: Aim 1. Define the molecular basis of insulator pairing specificity and its effects on chromosome architecture and gene expression. Aim 2. Determine the topological and functional consequences of manipulating endogenous insulators within and surrounding the eve locus. Aim 3. Test whether insulator pairing in cis competes with or facilitates insulator pairing in trans, and determine the resulting effects on gene expression.

基因转录的变化在癌症、大多数其他人类疾病的进展中非常重要， 在衰老过程中，以及在多细胞生物体发育的各个阶段。近年来它 染色体拓扑结构在基因调控中起着至关重要的作用， 流程.这种拓扑结构的主要决定因素涉及被称为绝缘体的调节DNA。工具 在果蝇中的可用性使得详细研究绝缘子作用和相互作用的机制成为可能， 真实的体内环境。本研究旨在研究基于染色质的基因调控机制， 绝缘体，包括最近在果蝇基因中发现的绝缘体，甚至跳过了。这些 研究将解决调节染色体拓扑结构的DNA如何执行其功能的基本问题。 在细胞核内的三维空间中发挥作用。他们将展示染色体结构如何与 通过整合绝缘子配对与增强子-启动子配对的拓扑结构来改变转录状态 交互.这些研究的一个独特贡献将是确定一个绝缘体配对代码， 预测和设计染色体结构及其对基因表达的影响。在 在哺乳动物中，已知绝缘子参与细胞类型特异性基因调控和关键发育调控。 决定，以及在肿瘤发生和遗传性人类疾病。因此，这些研究将直接 应用于人类疾病的研究。具体目标是： 目标1.定义绝缘子配对特异性的分子基础及其对染色体结构的影响 和基因表达。 目标二。确定操纵内部内源性绝缘体的拓扑和功能后果 并且围绕Eve轨迹。 目标3。测试顺式中的绝缘子配对是否与反式中的绝缘子配对竞争或促进其配对， 确定对基因表达的影响。