Stress regulation of RNA metabolism

RNA代谢的应激调节

• 批准号: 7156073
• 负责人: DANIEL W NEEF
• 金额: $ 4.88万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 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/7156073
• 关键词: SDS polyacrylamide gel electrophoresis; environmental stressor; fibroblasts; fungal genetics; fungal proteins; gene induction /repression; genetic regulation; genetic transcription; glutathione transferase; heat shock proteins; intermolecular interaction; microarray technology; nucleic acid metabolism; nucleic acid probes; postdoctoral investigator; protein binding; protein structure function; transcription factor

DESCRIPTION (provided by applicant): The ability to cope with environmental stress 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-alpha and c-fos. Interestingly, mammalian Tis11b, also a homologue of TTP is, like Cth1, 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 Tis11b is HSF-dependent, and to characterize mRNAs down regulated by Tisl Ib during stress and explore their role in stress-responses. Lastly, I aim to elucidate if mammalian TTP and Tis11b 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 Cth1, TTP and Tis11b on mRNA half-life.

描述（由申请人提供）：应对环境压力的能力对于细胞存活至关重要，迫使生物体开发出复杂的机制，通过这些机制，通过保护细胞成分免受应力诱导的损伤。不幸的是，对于响应压力而发生的mRNA代谢中发生的变化知之甚少。然而，酿酒酵母的最新证据表明，热休克因子（HSF）激活了可能参与mRNA代谢的几种基因的表达。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（是），其中包含TNF-Alpha和c-fos等mRNA。有趣的是，像CTH1一样，哺乳动物Tis11b也是由CTH1的同源物，尽管不知道这种诱导是否依赖于HSF。这些发现共同表明在细胞应激反应中介导的mRNA降解起作用。该建议旨在阐明压力期间mRNA代谢中发生的变化以及这些变化在细胞对压力反应中的重要性。具体而言，我建议确定CTHL在介导的降解中的作用在热应激过程中含有mRNA，探索是否依赖于HSF的压力依赖性诱导HSF依赖性，并表征由TISL IB调节的mRNA在压力过程中的作用并探索其在应力反应中的作用。最后，我的目标是阐明在细胞应激时期，在酵母中表达的哺乳动物TTP和TIS11b是否优先利用5'-3'mRNA衰变途径。这些问题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相互作用研究以及生化实验来探讨这些问题，以确定CTH1，TTP和TIS11B对mRNA半衰期的全局影响。