Stress regulation of RNA metabolism

RNA代谢的应激调节

基本信息

批准号：
7265294
负责人：
DANIEL W NEEF
金额：
$ 5.04万
依托单位：
DUKE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2007
资助国家：
美国
起止时间：
2007-01-01 至 2008-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7265294
关键词：
Address Binding Binding Proteins Binding Sites Biochemical Biological Assay Cell Cycle Arrest Cells Cellular Stress Cellular Stress Response Complex Condition Consensus Creativeness Data Elements Exonuclease Experimental Designs FOS gene Fellowship Funding Gene Expression Genes Growth Half-Life Heat Stress Disorders Heat shock proteins Heat-Shock Response Heating Heavy Metals Homologous Gene Human In Vitro Indium Individual Infection Mammals Mediating Messenger RNA Metabolism Microarray Analysis Modeling Names Organism Paper Pathway interactions Peer Review Physiological Process Productivity Protein Binding Proteins Publications Publishing RNA RNA-Protein Interaction Recovery Regulation Research Research Proposals Risk Role Saccharomyces cerevisiae Signal Transduction Stress Students TIS1 Testing Text Time Western Blotting Work Yeasts Zinc Fingers biological adaptation to stress coping design improved in vivo mRNA Decay mRNA Transcript Degradation mutant promoter protein folding repaired research study response stress protein thymidine 5&apos -triphosphate

项目摘要

The ability to cope with environmentalstress is essential to cellular survival, forcing organisms to develop sophisticated mechanisms by which cellular components are protected from stress-induced damage. Unfortunately,only little is known about the changes that occur in mRNA metabolism in response to stress. However, recent evidence from S. cerevisiae has shown that Heat Shock Factor (HSF) activates the expression of several genes likely to be involved in mRNA metabolism. One such gene is CTH1, encoding a tandem-zinc-finger protein whose human homologue, TTP, has been shown to promote the degradation of AU-rich element (ARE) containing mRNAs like TNF-a and c-fos. Interestingly, mammalian Tisl Ib, also a homologue of TTP is, like Cthl, inducible by stress, though it is not known if this induction is HSF dependent. Together these findings strongly suggest a role for ARE-mediated mRNA degradation in cellular stress-responses. This proposal aims to elucidate the changes that occur in mRNA metabolism during stress and the importance of these changes in cellular responses to stress. Specifically, I propose to determine the role of Cthl in mediating the degradation of ARE containing mRNAs during heat stress, explore if stress-dependent induction of Tisl Ib is HSF-dependent, and to characterize mRNAs downregulated by Tisl Ib during stress and explore their role in stress-responses. Lastly, I aim to elucidate if mammalian TTP and Tisl Ib expressed in yeast preferentially utilize the 5'-3' mRNA decay pathway during times of cell stress. These questions will be explored through the use of microarray analysis, in vivo and in vitro protein-protein and protein-RNA interaction studies, and biochemical experiments to determine the global effects of Cthl, TTP and Tisl Ib on mRNA half-life.

应付环保剂的能力对于细胞生存至关重要，迫使生物发展成熟通过保护细胞成分免受应力诱导损伤的机制。不幸的是，只有很少知道的关于响应压力的mRNA代谢中发生的变化。但是，酿酒酵母的最新证据有表明热休克因子（HSF）激活了可能与mRNA代谢有关的几个基因的表达。这样的基因是CTH1，编码了串联锌指蛋白，其人类同源物TTP已被证明可以促进富含Au元素的降解（）包含TNF-A和C-FOS等mRNA。有趣的是，哺乳动物Tisl IB，也 TTP的同源物与CTHL一样，通过应力诱导，尽管尚不清楚这种诱导是否取决于HSF。一起这些发现强烈表明在细胞应激反应中介导的mRNA降解起作用。该提议的目的为了阐明压力期间mRNA代谢中发生的变化以及这些变化在细胞中的重要性对压力的反应。具体而言，我建议确定CTHL在中介降解中的作用热应激期间的mRNA，探索是否依赖于TISL IB的应力依赖性诱导HSF依赖性并表征mRNA 压力期间由TISL IB下调，并探索其在压力响应中的作用。最后，我的目标是阐明哺乳动物是否在细胞应激时期，在酵母中表达的TTP和TISL IB优先利用5'-3'mRNA衰变途径。这些将通过使用微阵列分析，体内和体外蛋白质 - 蛋白质和蛋白-RNA来探讨问题相互作用研究以及生化实验，以确定CTHL，TTP和TISL IB对mRNA半衰期的全局影响。