Functional dissection of Drosophila TFIID

描述(由申请人提供)：真核转录的激活依赖于调控蛋白与其目标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潜在机制，作为开发检测和减轻人类疾病的诊断和治疗分析的先决条件。这项研究背后的具体假设是，通用转录因子TFIID中的TBP相关因子(TAFs)在后生动物转录中发挥重要作用。这一假设是基于以下观察结果：1)TAFs的突变是隐性致死的，2)TAFs与激活剂相互作用，3.)TAFs介导TFIID与启动子的结合，4.TAFs介导组蛋白和GTF的合成后修饰；TAFs在各种生物过程中发挥着重要作用。基于这些结果，这一建议的主要实验重点是在后生动物基因组的背景下TAFs对于转录激活的功能重要性。具体目标是： 1.剖析TAF1活性在转录调控中的功能重要性。TAF1通过组蛋白和GTF的合成后修饰，与TBP相互作用，介导TFIID向启动子募集，在转录激活中发挥重要作用。我们将全基因组转录图谱与染色质免疫沉淀实验相关联，以阐明TAF1的各种活性在后生动物转录中的功能重要性。 2.剖析TAFs在果蝇转录激活中的功能重要性。果蝇基因的转录状态与TAFs的存在、TAF介导的蛋白质修饰、激活剂以及启动子上的RNA聚合酶II转录机制(GTM)的相关性将阐明TAFs在转录激活中的功能重要性。 3.剖析TAFs在体内执行生物过程中的作用。我们将把特定基因的转录与TAFs的存在、TAF介导的蛋白质修饰和基因启动子上的GTM相关联，以阐明TAFs在体型形成、细胞周期进展和表观激活中的功能重要性。

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.