Bacterial RNA thermometers: From individual examples to global analyses

细菌 RNA 温度计：从个体实例到全局分析

• 批准号: 244588142
• 负责人: Professor Dr. Franz Narberhaus
• 金额: --
• 依托单位: Lehrstuhl für Biologie der Mikroorganismen
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2013
• 资助国家: 德国
• 起止时间: 2012-12-31 至 2021-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/244588142?language=en
• 关键词: Bacterial; RNA; thermometers; individual; examples

Structured RNA regions influence every step in the gene expression program. Many bacterial heat shock and virulence genes are regulated by RNA thermometers (RNATs), which are molecular zippers that control translation initiation. They adopt a structure that blocks the ribosome binding site at low temperatures. A temperature upshift to 37°C (virulence genes) or higher (heat shock genes) destabilizes the structure and permits translation.In the previous funding period, we have studied various natural and synthetic RNATs in molecular detail. By using a combination of in vitro structure probing and next-generation sequencing we mapped the entire RNA structurome of the human pathogen Yersinia pseudotuberculosis at three different temperatures. We now want to take advantage of this comprehensive data resource and characterize two distinct types of regulation. (i) Following the initial characterization of more than 25 novel RNATs, the structure and physiological function of a number of these regulatory elements will be evaluated. We are particularly interested in the virulence factor CnfY, and components of the oxidative stress response and type III secretion system. (ii) The RNA structurome results suggest a regulatory concept, in which several small regulatory RNAs undergo temperature-dependent conformational changes that mask or unmask their target interaction region. We will further evaluate this observation and study its effect on target gene regulation. In a third work package, we plan to capitalize on a previously developed in vivo RNA structuromics approach with the goal to learn how RNAs fold in the living cell.

结构RNA区域影响基因表达程序的每一步。许多细菌的热休克和毒力基因受RNA温度计(RNAT)的调控，RNAT是控制翻译启动的分子拉链。它们采用了一种在低温下阻断核糖体结合部位的结构。温度升高到37℃(毒力基因)或更高(热激基因)会破坏结构的稳定，并允许翻译。在之前的资助期间，我们对各种天然和合成的RNAs进行了详细的分子研究。通过使用体外结构探测和下一代测序的组合，我们绘制了人类病原体假结核耶尔森菌在三种不同温度下的整个RNA结构。我们现在想要利用这一全面的数据资源，并描述两种不同类型的监管。(I)在初步鉴定了超过25种新的RNAT之后，将对其中一些调节元件的结构和生理功能进行评估。我们特别感兴趣的是毒力因子CNFy，以及氧化应激反应和III型分泌系统的组成部分。(Ii)RNA结构结果提出了一个调控概念，在这个概念中，几个小的调控RNA经历了依赖于温度的构象变化，掩盖或揭开了它们的目标相互作用区域。我们将进一步评估这一观察结果，并研究其对靶基因调控的影响。在第三个工作包中，我们计划利用之前开发的体内RNA结构学方法，目标是了解RNA如何在活细胞中折叠。