Regulation of DNA Damage Induced Genes by Yeast TAFIIs

酵母TAFII对DNA损伤诱导基因的调控

基本信息

批准号：
8847902
负责人：
JOSEPH C REESE
金额：
$ 2.48万
依托单位：
PENNSYLVANIA STATE UNIVERSITY, THE
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-01-01 至 2015-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8847902
关键词：
Affect Animals Attention Binding Biochemical Biochemistry Biological Assay Birth Cardiovascular system Cell Proliferation Cells Cessation of life ChIP-on-chip Collaborations Complex DNA Damage Data Defect Development Disease Disease Resistance Elongation Factor Eukaryota Eukaryotic Cell Event Funding Gene Expression Gene Expression Regulation General Transcription Factors Genes Genetic Genetic Transcription Genome Genomics Genotoxic Stress Goals Health Histones Human In Vitro Length Link Location Maps Measures Methods Methylation MicroRNAs Mutation Physiological Play Process Proteins Publishing RNA RNA Polymerase II RNA Sequences Regulation Regulator Genes Reporter Genes Research Resistance Role Site Source Structure Syndrome Techniques Transcript Transcription Elongation Transcription Factor TFIID Transcription Initiation Work Yeast Model System Yeasts biological adaptation to stress density fascinate genome wide association study human disease in vivo insight mRNA Transcript Degradation mutant novel programs protein complex transcription factor transcription factor S-II

项目摘要

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, and determining how these events are coordinated is central to understanding gene expression. The yeast Ccr4-Not complex was originally described as a regulator of the general transcription factor TFIID. Remarkably, it has now been implicated in multiple aspects of gene regulation including mRNA degradation, translational silencing, histone methylation and protein ubiquitylation. Its myriad of functions is quite fascinating and suggests it regulates gene expression from the birth to death of a gene product. Our study of TFIID and DNA damage inducible genes led us to characterize this complex, as it appears to play a role in regulating DNA damage stress responses. Over the last funding period we established it as a bona-fide elongation factor that directly activates arrested RNA polymerase II (RNAPII). Interestingly, Ccr4-Not appears to function via a mechanism unique from other elongation factors characterized to date; thus, analyzing its function has the potential to reveal novel insights into how factors regulate RNAPII. We have a developed a number of powerful assays to fully exploit the biochemistry, powerful genetics and genomics of yeast to characterize its mechanism of action and the role it plays in gene expression in vivo. The goals of the studies described in this proposal are to investigate the structure of arrested elongation complexes bound by Ccr4-Not to gain a greater understanding of how it promotes elongation; to explore its collaboration with other elongation factors to regulate RNAPII activity; to analyze the physiological functions of the complex in vivo by mapping changes in RNAPII arrest across the genome in Ccr4-Not mutants. The long term goals of this project are to understand how this multi-functional complex regulates multiple steps in gene regulation to maintain normal development and the integrity of the genome in eukaryotic cells.

描述（由申请人提供）：基因表达的调节是细胞中的一个基本过程，并且此过程的改变与人类的许多疾病状态有关。它在多个层面上进行调节，并确定这些事件的协调方式对于理解基因表达至关重要。酵母CCR4不合期的复合物最初被描述为一般转录因子TFIID的调节剂。值得注意的是，它与基因调节的多个方面有关，包括mRNA降解，转化沉默，组蛋白甲基化和蛋白质泛素化。它的无数功能令人着迷，这表明它从基因产物的诞生到死亡。我们对TFIID和DNA损伤诱导基因的研究使我们表征了这一复合物，因为它似乎在调节DNA损伤应力反应中起作用。在最后一个资金期间，我们将其确定为一个真正的伸长因子，直接激活被捕的RNA聚合酶II（RNAPII）。有趣的是，CCR4-----------------迄今为止的其他伸长因子与其他伸长因子所独有的机制起作用。因此，分析其功能有可能揭示有关因素如何调节RNAPII的新见解。我们开发了许多有力的测定方法，可以充分利用生物化学，强大的遗传学和酵母基因组学来表征其作用机理及其在体内基因表达中所扮演的作用。该提案中描述的研究的目标是研究由CCR4约束的被捕伸长率复合物的结构，而不是对其促进延伸的方式有更深入的了解；探索其与其他伸长因素的合作，以调节RNAPII活动；通过绘制CCR4-不突变体基因组的RNAPII停滞的变化来分析复合体体内的生理功能。该项目的长期目标是了解这种多功能复合物如何调节基因调节的多个步骤，以维持真核细胞中基因组的正常发育和完整性。