Insulators: Determinants of chromosome topology and regulatory interactions

基因转录的变化在癌症的进展中很重要，在大多数其他人类疾病中，以及 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 染色体拓扑学在基因调控中起着至关重要的作用，就像其他核一样。 流程。这种拓扑的主要决定因素涉及被称为绝缘体的调节性DNA。工具 在果蝇中的可用使详细研究绝缘体的作用和相互作用的机制成为可能，在 在活体环境中是正确的。这项建议是为了研究基于染色质的基因调控机制，包括 绝缘体，包括最近在果蝇基因中发现的那些绝缘体，甚至跳过了。这些 研究将解决基本问题，即调节染色体拓扑的调控DNA如何执行其 在核内的三个维度上起作用。他们将展示染色体结构如何与 整合绝缘子配对和增强子-启动子拓扑的交替转录状态 互动。这些研究的独特贡献将是确定可以使用的绝缘子配对代码 预测和设计染色体构型及其对基因表达的影响。在……里面 哺乳动物，绝缘体已知参与细胞类型的特定基因调节和关键的发育 决定，以及在肿瘤发生和遗传性人类疾病。因此，这些研究将直接具有 在人类疾病研究中的应用。具体目标是： 目的1.明确绝缘子配对专一性的分子基础及其对染色体构型的影响 和基因表达。 目标2.确定控制内部绝缘子的拓扑和功能后果 围绕着夏娃轨迹。 目标3.测试顺式绝缘子配对是否竞争或促进换向绝缘子配对，以及 确定由此产生的对基因表达的影响。

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.