Regulation of DNA Damage Induced Genes by Yeast TAFIIs

酵母TAFII对DNA损伤诱导基因的调控

• 批准号: 6825873
• 负责人: JOSEPH C REESE
• 金额: $ 22.44万
• 依托单位: PENNSYLVANIA STATE UNIVERSITY, THE
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-01-01 至 2007-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6825873
• 关键词: DNA damage; DNA directed RNA polymerase; antibody; chromatin; enzyme inhibitors; fungal genetics; gene induction /repression; gene mutation; genetic promoter element; genetic regulation; genetic transcription; histones; intermolecular interaction; laboratory rabbit; nucleosomes; protein binding; ribonucleotide reductase; transcription factor

DESCRIPTION (provided by applicant): Gene regulation plays a central role in all cellular functions, and alterations in this process have been associated with a number of developmental defects and disease states in humans. Initiation of gene expression requires changes to the chromatin environment of genes and the formation of the pre-initiation complex (PIC) at the promoter. The PIC is formed by the ordered recruitment of multiple general transcription factors (GTFs). Advances over the last five years indicate that chromatin is not simply an inert barrier to transcription, but may also play an important function in the recruitment of specific transcription factors to sites within the genome. Further, the distinction between the functions of GTFs and the chromatin remodeling machinery has been blurred by the discovery that certain GTFs have intrinsic chromatin modifying activities, or can recruit them to promoters. The overall goal of this proposal is to understand how chromatin remodeling complexes and Principal Investigator C components are recruited to repressed promoters, and how coordination of the chromatin remodeling and transcription steps occurs. We are using the DNA damage-responsive ribonucleotide reductase 3 gene (RNR3) of budding yeast to explore the functions of the TFIID complex, which is composed of the TATA-binding protein and 14 associated factors called TAFs. Our efforts over the last funding period revealed that TFIID is required for the recruitment of the SWI/SNF nucleosome remodeling complex and the remodeling of nucleosomes at the promoter of RNR3. The objectives of this proposal are to identify the features of RNR3 that confer its dependence upon TAFIIS and the general transcription machinery for nucleosome remodeling. The aims of this proposal will address the role of transcription and the core promoter, identify the components of the pre-initiation complex (PIC) required for SWI/SNF recruitment, and analyze the interactions between PIC components, SWI/SNF and chromatin in an in vitro system. Further, we will explore the contributions of the "histone code" in the remodeling and recruitment of transcription factors to RNR3 using genetic strategies to alter core histone modification states and mutating histone tails. These studies will define mechanisms of transcription factor recruitment, chromatin remodeling and the functions of the TFIID complex that will be applicable to other eukaryotic model systems.

描述（由申请人提供）：基因调控在所有细胞功能中起核心作用，并且该过程中的改变与人类的许多发育缺陷和疾病状态相关。基因表达的起始需要基因的染色质环境的改变和在启动子处的前起始复合物（PIC）的形成。PIC是由多个通用转录因子（GTF）有序募集而形成的。过去五年的研究进展表明，染色质不仅是转录的惰性屏障，而且可能在将特定转录因子募集到基因组内的位点中发挥重要作用。此外，由于发现某些GTF具有内在的染色质修饰活性，或者可以将它们募集到启动子中，GTF的功能与染色质重塑机制之间的区别已经变得模糊。本提案的总体目标是了解染色质重塑复合物和主要研究者C组分是如何被招募到受抑制的启动子中的，以及染色质重塑和转录步骤是如何协调的。我们正在使用芽殖酵母的DNA损伤响应性核糖核苷酸还原酶3基因（RNR 3）来探索TFIID复合物的功能，该复合物由TATA结合蛋白和14种称为TAFs的相关因子组成。我们在上一个资助期的努力表明，TFIID是招募SWI/SNF核小体重塑复合物和在RNR 3启动子处重塑核小体所必需的。本提案的目的是确定RNR 3的功能，赋予其依赖于TAFIIS和核小体重塑的一般转录机制。本提案的目的是解决转录和核心启动子的作用，确定SWI/SNF招募所需的前起始复合物（PIC）的组件，并分析PIC组件，SWI/SNF和染色质之间的相互作用在体外系统。此外，我们将探索的贡献，“组蛋白密码”在重塑和招募的转录因子RNR 3使用遗传策略，改变核心组蛋白修饰状态和突变组蛋白尾部。这些研究将定义转录因子募集，染色质重塑和TFIID复合物的功能，将适用于其他真核模型系统的机制。