Biochemical, Genomic and Computational Analysis of Transcriptional Repression

项目摘要 转录抑制因子是后生动物中转录激活因子的必要平衡物 发展转录抑制的机制已被深入研究，但我们缺乏 对阻遏物如何在基因组中的许多靶点上在机械水平上起作用的重要见解。特别是， 转录辅阻遏物的协调募集可以对染色质结构产生不同的影响 和修改，我们仍然缺乏对许多变化的功能意义的见解， 在经济学研究中衡量。为了深入了解真核生物的转录调控机制，我们 使用果蝇胚胎的自然环境来识别发育过程中的基本生化过程， 设置，其中差异基因表达用于驱动特定细胞的发育命运， 组织中在这项提议中，1）我们将使用我们实验室开发的全基因组方法来识别直接 由一组五种内源性转录因子指导的基于染色质的生物化学过程 监管机构通过短期和长期机制进行抑制。2)我们将研究在体内活性的 野生型和突变阻遏复合物，以确定不同的转录辅阻遏物的贡献 Groucho和CtBP，测试它们对染色质中定量和/或定性效应的贡献 修饰和基因调控。3)为了确定转录抑制因子的顺式调节环境， 作用于不同的增强子，我们将定量测量和数学模型的输出， 增强子，以揭示特定类别的阻遏物相互作用的基本定量特性， 激活因子-即这些蛋白质与许多靶基因相互作用的共同“规则”。三 相互关联的目标将提供预测工具，用于解释基因组顺式调控内容的基因组顺式调控内容。 后生动物基因组，为高等动物的进化和疾病研究提供必要的基础。 真核生物

Project Summary Transcriptional repressors represent the necessary counterweight to transcriptional activators in metazoan development. The mechanisms of transcriptional repression have been intensively investigated, but we lack crucial insights into how repressors act at a mechanistic level across many targets in the genome. In particular, the coordinated recruiting of transcriptional co-repressors can generate diverse effects on chromatin structure and modification, and we still lack insights on the functional significance of many changes that can be measured in `omics studies. To develop key insights into eukaryotic transcriptional regulatory mechanisms, we use the natural setting of the Drosophila embryo to identify basic biochemical processes in a developmental setting, where differential gene expression is used to drive the developmental fate of particular cells and tissues. In this proposal, 1) we will use genome-wide methods developed in our laboratory to identify direct biochemical, chromatin-based processes that are directed by a set of five endogenous transcriptional regulators that repress through short-range and long-range mechanisms. 2) We will study the in vivo activity of wild-type and mutant repression complexes to identify the contributions of distinct transcriptional co-repressors Groucho and CtBP, testing their contributions to quantitative and/or qualitative effects in chromatin modifications and gene regulation. 3) To identify the cis-regulatory context in which transcriptional repressors act on different enhancers, we will quantitatively measure and mathematically model the output of specific enhancers to uncover the fundamental quantitative properties of specific classes of repressors interacting with activators – i.e. the common “rules” by which these proteins interact on many target genes. The three interrelated aims will provide predictive tools for interpretation of genomic cis regulatory content of the metazoan genome, providing essential underpinnings for studies of evolution and disease in higher eukaryotes.