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(申请人提供)：基因调控在所有细胞功能中起着核心作用，这一过程中的变化与人类的一些发育缺陷和疾病状态有关。基因表达的启动需要改变基因的染色质环境和在启动子上形成预启动复合体(PIC)。PIC是由多个通用转录因子(GTF)有序招募而成。过去五年的研究进展表明，染色质不仅是转录的惰性屏障，还可能在将特定转录因子招募到基因组内的位置上发挥重要作用。此外，GTF的功能和染色质重塑机制之间的区别已经被某些GTF具有内在的染色质修饰活性或可以招募它们为启动子的发现所模糊。该提案的总体目标是了解染色质重塑复合体和主要调查者C组分如何被招募到抑制启动子，以及染色质重塑和转录步骤是如何发生协调的。我们正在使用芽期酵母的DNA损伤反应核糖核苷酸还原酶3基因(RNR3)来探索TFIID复合体的功能，该复合体由TATA结合蛋白和14个称为TAFs的相关因子组成。我们在上一个资金阶段的工作表明，TFIID对于SWI/SNF核小体重塑复合体的招募和RNR3启动子的核小体重塑是必需的。这项建议的目的是确定RNR3的特征，使其依赖于TAFIIS和核小体重塑的一般转录机制。该建议的目的是解决转录和核心启动子的作用，确定SWI/SNF招募所需的预起始复合体(PIC)的成分，并在体外系统中分析PIC成分、SWI/SNF和染色质之间的相互作用。进一步，我们将通过改变核心组蛋白修饰状态和突变的组蛋白尾巴的遗传策略，探索“组蛋白密码”在转录因子对RNR3的重塑和招募中的作用。这些研究将确定转录因子招募、染色质重塑的机制以及TFIID复合体的功能，这些机制将适用于其他真核模型系统。