REGULATION OF DNA DAMAGE INDUCED GENES BY YEAST TAFIIS

酵母 TAFIIS 对 DNA 损伤诱导基因的调控

• 批准号: 6418015
• 负责人: JOSEPH C REESE
• 金额: $ 4.95万
• 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 2003-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6418015
• 关键词: DNA binding protein; DNA damage; DNA footprinting; DNA repair; acyltransferase; biological signal transduction; chromatin; crosslink; fungal genetics; gene induction /repression; genetic promoter element; genetic regulation; genetic regulatory element; genetic transcription; histones; immunoprecipitation; northern blottings; nucleosomes; phosphorylation; protein kinase; ribonucleotide reductase; tissue /cell culture; transcription factor

Cancer, birth defects and aging are all thought to result from of an accumulation of genetic damage. Cells respond to the challenge of DNA damage by activating a protein kinase cascade that causes cell cycle arrest and the induction of repair genes. Alterations in either of these responses lead to human disease. Kinases in these pathways have been identified, however, their transcription factor targets have remained elusive. We have identified potential targets. Through our genetic study of the TATA-box binding protein associated factors (yTAFIIS) from the budding yeast Saccharomyces cerevisiae, we have discovered that yTAFIIS are required for the cellular response to DNA damage and damage- induced transcription. This is a significant discovery, because the in vivo functions of TAFIIS are largely unknown. Biochemical studies have implicated them as being important for the control of transcription from all promoters, and they are thought to do so by acting as coactivators and promoter selectivity factors. However, analysis of TAFII mutants in vivo in both yeast and mammalian systems have challenged this model. Mutation of multiple yeast TAFIIS, and mutation of the largest mammalian TAFII, do not lead to global defects in transcription, but lead to cell cycle defects and alterations in transcription of a select number of genes. Previous work identified two classes of genes that are dependent upon TAF145 for transcription. Most surprisingly, TAF145 determinants are not activator binding sites, but core promoter elements, leaving the issue of what signals are acting through TAFIIS an open question. We have found that the transcription of DNA damage response genes requires multiple TAFII subunits, which makes their regulation distinct from the previously identified gene classes. This proposal will extend these preliminary observations by characterizing the role of TAFIIS in the regulation of DNA damage responsive genes and explore the signaling pathways that mediate their activities. These studies will identify the determinants of TAFII-dependent genes, the cellular signals controlling TAFII function, and attempt to establish TAFIIS as the transcription factor targets of the DNA damage protein kinase cascade. This work will impact the fields of transcription, DNA damage and cell cycle control.

癌症，出生缺陷和衰老都被认为是由 遗传损害的积累。 细胞应对DNA的挑战 通过激活导致细胞周期的蛋白激酶级联反应来损害 逮捕和诱导修复基因。其中任何一个的改变 反应导致人类疾病。这些途径中的激酶已经 然而，确定其转录因子靶标仍然存在 难以捉摸。 我们已经确定了潜在目标。 通过我们的遗传 从研究中研究TATA-box结合蛋白相关因素（YTAFII） 酿酒酵母的萌芽酵母菌，我们发现 Ytafiis是对DNA损伤和损伤的细胞反应所必需的 诱导转录。 这是一个重大发现，因为 Tafiis的体内功能在很大程度上未知。生化研究已有 暗示它们对于控制转录的重要 所有发起人，他们被认为是通过充当共激活者而这样做的 和启动子选择性因素。 但是，分析TAFII突变体 酵母菌和哺乳动物系统中的体内都对该模型提出了挑战。 多种酵母Tafiis的突变和最大的哺乳动物的突变 tafii，不会导致转录的全局缺陷，而是导致细胞 选择数量数量的转录的周期缺陷和变化 基因。 以前的工作确定了两类基因 在TAF145进行转录时。 最令人惊讶的是，TAF145决定因素 不是激活器绑定位点，而是核心启动子元素，留下 问题是什么信号通过tafiis做出的一个悬而未决的问题。我们 发现DNA损伤反应基因的转录需要 多个TAFII亚基，这使得其调节与 先前确定的基因类。 该提议将延长这些 通过表征Tafiis在 调节DNA损伤响应基因并探索信号传导 调节他们的活动的途径。 这些研究将确定 TAFII依赖性基因的决定因素，控制的细胞信号 TAFII功能，并尝试将Tafiis建立为转录 DNA损伤蛋白激酶级联反应的因子靶标。 这项工作将 影响转录，DNA损伤和细胞周期控制的场。