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生存至关重要，迫使生物体发展出复杂的机制来保护细胞成分免受压力引起的损伤。不幸的是，人们对 mRNA 代谢因压力而发生的变化知之甚少。然而，最近来自酿酒酵母的证据表明，热休克因子 (HSF) 激活了几个可能参与 mRNA 代谢的基因的表达。其中一个基因是 CTH1，它编码一种串联锌指蛋白，其人类同源物 TTP 已被证明可以促进含有 TNF-α 和 c-fos 等 mRNA 的 AU 富集元件 (ARE) 的降解。有趣的是，哺乳动物的 Tis11b（也是 TTP 的同源物）与 Cth1 一样，可被应激诱导，尽管尚不清楚这种诱导是否依赖于 HSF。这些发现共同有力地表明了 ARE 介导的 mRNA 降解在细胞应激反应中的作用。该提案旨在阐明应激期间 mRNA 代谢发生的变化以及这些变化在细胞对应激反应中的重要性。具体来说，我建议确定 Cthl 在介导热应激期间含有 mRNA 的 ARE 降解中的作用，探索 Tis11b 的应激依赖性诱导是否是 HSF 依赖性的，并表征应激期间 Tisl Ib 下调的 mRNA 并探索其作用在压力反应中。最后，我的目的是阐明酵母中表达的哺乳动物 TTP 和 Tis11b 在细胞应激期间是否优先利用 5'-3' mRNA 衰减途径。这些问题将通过使用微阵列分析、体内和体外蛋白质-蛋白质和蛋白质-RNA相互作用研究以及生化实验来探讨，以确定Cth1、TTP和Tis11b对mRNA半衰期的总体影响。