Functional dissection of Drosophila TFIID

果蝇 TFIID 的功能解剖

• 批准号: 6969722
• 负责人: FRANK U SAUER
• 金额: $ 29.91万
• 依托单位: UNIVERSITY OF CALIFORNIA RIVERSIDE
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-08-01 至 2009-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6969722
• 关键词: DNA directed RNA polymerase; Drosophilidae; RNA interference; biotechnology; cell cycle; chromatin immunoprecipitation; eukaryote; functional /structural genomics; gene expression; gene expression profiling; gene induction /repression; genetic transcription; in situ hybridization; laboratory rabbit; microarray technology; molecular shape; polymerase chain reaction; transcription factor

DESCRIPTION (provided by applicant): Activation of eukaryotic transcription depends on a refined interplay of regulatory proteins with their target genes. Our long-term goal is to elucidate the mechanisms underlying regulation of metazoan gene expression as a prerequisite for the development of diagnostic and therapeutic assays that detect and attenuate human diseases. The specific hypothesis behind the proposed research is that the "TBP-associated factors" (TAFs) in the general transcription factor TFIID play an important role in metazoan transcription. That hypothesis is based on the observations that 1) mutations in TAFs are recessive lethal, 2.) TAFs interact with activators, 3.) TAFs mediate binding of TFIID to promoters, 4.) TAFs mediate the postsynthetic modification of histones and GTFs, and 5.) TAFs play an important role in various biological processes. Based on these results the main experimental focus of this proposal is on the functional importance of TAFs for transcription activation in the context of a metazoan genome. The specific aims are to: 1. To dissect the functional importance of TAF1 activities in regulation of transcription. TAF1 plays an important role in transcription activation by postsynthetic modification of histones and GTFs, interacting with TBP, mediating the recruitment of TFIID to promoters. We will correlate genome-wide transcription profiling with chromatin immunoprecipitation experiments to elucidate the functional importance of the various activities in TAF1 in metazoan transcription. 2. To dissect the functional importance of TAFs in activation of transcription in Drosophila. The correlation of the transcription status of Drosophila genes with the presence of TAFs, TAF-mediated protein modifications, activators, and the RNA polymerase II transcription machinery (GTM) at promoters will elucidate the functional importance of TAFs in transcription activation. 3. To dissect the role of TAFs in the execution of biological processes in vivo. We will correlate the transcription of specific genes with the presence of TAFs, TAF-mediated protein modifications, and the GTM at the promoters of the genes to elucidate the functional importance of TAFs for body pattern formation, cell cycle progression, and epigemetic activation.

描述（由申请人提供）：真核转录的激活取决于调节蛋白与其靶基因的精细相互作用。我们的长期目标是阐明后生动物基因表达调控的机制，这是开发检测和减轻人类疾病的诊断和治疗方法的先决条件。该研究背后的具体假设是通用转录因子TFIID中的“TBP相关因子”（TAF）在后生动物转录中发挥重要作用。该假设基于以下观察结果：1) TAF 的突变是隐性致死的，2.) TAF 与激活剂相互作用，3.) TAF 介导 TFIID 与启动子的结合，4.) TAF 介导组蛋白和 GTF 的合成后修饰，以及5.) TAF 在各种生物过程中发挥着重要作用。基于这些结果，该提案的主要实验重点是 TAF 对于后生动物基因组中转录激活的功能重要性。具体目标是： 1. 剖析 TAF1 活性在转录调节中的功能重要性。 TAF1 通过组蛋白和 GTF 的合成后修饰、与 TBP 相互作用、介导 TFIID 募集至启动子而在转录激活中发挥重要作用。我们将把全基因组转录分析与染色质免疫沉淀实验联系起来，以阐明 TAF1 各种活性在后生动物转录中的功能重要性。 2. 剖析 TAF 在果蝇转录激活中的功能重要性。果蝇基因的转录状态与 TAF、TAF 介导的蛋白质修饰、激活剂和启动子处的 RNA 聚合酶 II 转录机制 (GTM) 存在的相关性将阐明 TAF 在转录激活中的功能重要性。 3. 剖析 TAF 在体内生物过程执行中的作用。我们将把特定基因的转录与 TAF 的存在、TAF 介导的蛋白质修饰以及基因启动子处的 GTM 关联起来，以阐明 TAF 对身体模式形成、细胞周期进展和表观遗传激活的功能重要性。