Molecular Analysis of Transcriptional Repression

转录抑制的分子分析

基本信息

批准号：
7545912
负责人：
David N Arnosti
金额：
$ 30.17万
依托单位：
MICHIGAN STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
1998
资助国家：
美国
起止时间：
1998-01-01 至 2010-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7545912
关键词：
Animals Binding Bioinformatics Biological Biological Assay Biological Models Blastoderm Body Patterning C-terminal C-terminal binding protein Complex DNA DNA Binding Domain Development Developmental Gene Disease Drosophila genus Effectiveness Elements Embryo Engineered Gene Enhancers Event Evolution Gene Expression Process Gene Expression Regulation Genes Genetic Enhancer Element Genomics Goals Histone Deacetylase Laboratories Laboratory Research Lead Maps Modeling Molecular Molecular Analysis Molecular Genetics Nuclear Hormone Receptors Nucleic Acid Regulatory Sequences Orphan Pathway interactions Pattern Peptides Physiological Play Process Property Protein C Proteins RNA Splicing Regulation Regulatory Element Reporter Genes Repression Research Role Signal Transduction Surface Testing Transcription Coactivator Transcription Repressor/Corepressor Transcriptional Regulation Variant Work Zinc Fingers base chromatin immunoprecipitation cofactor contextual factors design fly gene repression insight mathematical model novel programs promoter response stoichiometry tool

项目摘要

Transcriptional repression plays a key role in regulation of gene expression in development and in disease processes. To understand conserved regulatory processes in animals, we have determined both the molecular workings and physiological relevance of transcriptional represser proteins that play key roles in Drosophila development. Knirps is a key regulator of the even-skipped gene, acting on separate enhancer elements via a short-range mechanism. We have discovered how Knirps and related repressers function via distinct repression mechanisms, involving the evolutionary conserved CtBP corepressor protein and a histone deacetylase Rpd3, and we have developed novel insights into the design of cis regulatory elements that bind these repressers. We have identified key aspects of a "cisregulatory grammar" that predicts how represser proteins can function on defined regulatory elements, laying the groundwork for mathematical and bioinformatic analysis of endogenous enhancers. This work will allow us to apply Drosophila-derived information to general analysis of metazoan cis regulatory element design and evolution. 1. We will quantitatively "map" regulatory surfaces of genes controlled by transcriptional repressors, and use potential function-based mathematical models to predict the regulatory properties of novel enhancers. 2. We will elucidate of mechanisms of short- and long-range repressors using chromatin immunoprecipitation to identify molecular events associated with repression of embryonic reporter genes. 3. We will assess the function of corepressors we discovered associated with the Knirps represser complex, in particular the evolutionary conserved Groucho corepressor, whose role in short-range repression was previously unrecognized. 4. We will identify the physiological significance of conserved residues and splice-variants of the CtBP corepressor, using whole-animal assays to identify the biological significance of the proteins' activities. These aims combine empirical and modeling efforts to obtain a "bottoms up" understanding of transcriptional repression. Our long-term goal is to understand molecular activities of transcriptional control proteins and DNA regulatory sequences that are central to gene expression processes important in development and disease.

转录抑制在发育和疾病中基因表达的调节中起着关键作用流程。为了了解动物中保守的调节过程，我们确定了转录抑制蛋白的分子运作和生理相关性，在果蝇发育。 Knirps 是偶数跳跃基因的关键调节因子，作用于单独的增强子通过短程机制的元素。我们已经发现 Knirps 和相关抑制器如何通过不同的抑制机制，涉及进化保守的 CtBP 辅阻遏蛋白和组蛋白脱乙酰酶 Rpd3，我们对顺式调控元件的设计有了新的见解结合这些阻遏物。我们已经确定了“顺调控语法”的关键方面，它预测了如何阻遏蛋白可以在特定的调控元件上发挥作用，为数学和内源增强子的生物信息分析。这项工作将使我们能够应用果蝇衍生的后生动物顺式调控元件设计和进化的一般分析信息。 1. 我们将定量“绘制”转录抑制因子控制的基因的调控表面，以及使用基于势函数的数学模型来预测新型增强剂的调节特性。 2. 我们将利用染色质阐明短程和长程阻遏物的机制免疫沉淀来识别与胚胎报告基因抑制相关的分子事件。 3. 我们将评估我们发现的与 Knirps 抑制子相关的辅抑制子的功能复合物，特别是进化保守的 Groucho 辅阻遏物，其在短程中的作用镇压以前未被认识到。 4. 我们将鉴定CtBP的保守残基和剪接变体的生理意义辅阻遏物，使用整体动物测定来鉴定蛋白质活性的生物学意义。这些目标结合了实证和建模工作，以获得对以下内容的“自下而上”的理解：转录抑制。我们的长期目标是了解转录控制的分子活动对基因表达过程至关重要的蛋白质和 DNA 调控序列发育和疾病。