Regulation of DNA Damage Induced Genes by Yeast TAFIIs

描述（由申请人提供）：基因表达的调控是细胞中的一个基本过程，该过程的改变与人类的许多疾病状态有关。它在多个水平上受到调控，需要高度协调和整合的事件，包括染色质重塑、起始、延伸、加工和mRNA的最终破坏。确定这些事件如何协调是理解基因表达的核心。本提案的总体目标是了解转录因子复合物如何协调基因调控的多个步骤。本提案将研究ccr4 -不复杂的功能，利用酵母强大的遗传系统。Ccr4-Not复合体与一般转录因子复合体TFIID有许多联系，特别是TAFII亚基。提出了遗传、生化和分子方法。目的1将描述Ccr4- Not转录复合体在转录过程中的功能。该复合体显示了与TFIID、一般转录机制和延伸因子的多种物理和遗传联系。它在转录周期多个阶段的作用将以RNR3作为模型基因进行研究。目的2将确定Ccr4-Not的靶向机制。它调节基因的一个子集，似乎不是细胞中所有转录都普遍需要的。我们将探索其在RNR3上的招募和功能所需的DNA序列和转录因子。目标3将采用基于基因组学的方法来鉴定新的ccr4 -非依赖性基因，并鉴定指定该复合体调控的基因组特征。这些方法的交叉将允许更好地理解转录中多个步骤的协调。完成这些目标将：(1)揭示基因表达的多个步骤是如何协调和调节的；(2)确定与人类疾病相关的高度保守的真核转录因子复合体的功能；(3)为确定如何标记特异性mrna以进行转录后控制奠定基础。

DESCRIPTION (provided by applicant): The regulation of gene expression is a fundamental process in cells, and alterations in this process have been linked to numerous disease states in humans. It is regulated at multiple levels, requiring highly coordinated and integrated events including chromatin remodeling, initiation, elongation, processing and ultimately the destruction of mRNA. Determining how these events are coordinated is central to understanding gene expression. The overall objective of this proposal is to understand how transcription factor complexes coordinate multiple steps in gene regulation. This proposal will examine the functions of the Ccr4-Not complex, exploiting the powerful genetic system of yeast. The Ccr4-Not complex has many ties to the general transcription factor complex TFIID, particularly the TAFII subunits. Genetic, biochemical and molecular approaches are proposed. Aim 1 will characterize the functions of the Ccr4- Not transcription complex in the process of transcription. This complex displays multiple physical and genetic links to TFIID, the general transcription machinery and elongation factors. Its role in multiple stages in the transcription cycle will be examined using RNR3 as a model gene. Aim 2 will identify the targeting mechanism for Ccr4-Not. It regulates a subset of genes, and appears not be universally required for all transcription in the cell. We will explore the DNA sequence and transcription factor requirements for its recruitment and function at RNR3. Aim 3 will employ genomics based approaches to identify novel Ccr4-Not dependent genes and identify the features of the genome that specifies regulation by this complex. The intersection of these approaches will allow for a greater understanding of the coordination of multiple steps in transcription. The completion of these aims will: (1) uncover how multiple steps in gene expression are coordinated and regulated; (2) define the functions of a highly conserved eukaryotic transcription factor complex implicated in human disease; (3) lay the groundwork for identifying how specific mRNAs are marked for post-transcriptional control. Project Narrative - Relevance: Cells require precise control over the expression of their genes, and numerous human diseases and syndromes are caused by disturbances in gene expression. A multitude of transcription factors coordinate their activities to regulate this important process. The goal of the work described here is to understand how transcription factors control the level of gene expression; thus, completion of this work is directly relevant to human health.