Structure, molecular mechanism and biotechnological application of RNA thermometers

RNA温度计的结构、分子机制及生物技术应用

• 批准号: 5436052
• 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: 2004
• 资助国家: 德国
• 起止时间: 2003-12-31 至 2007-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/5436052?language=en
• 关键词: Structure; molecular; mechanism; biotechnological; application

Direct sensing of environmental cues has only recently been recognized as an intrinsic property of certain mRNAs. Conformational changes in the 5`-untranslated region (5`-UTR) of sensory RNA molecules are predicted to control either transcription or translation of downstream genes. We are currently studying the RNA thermometer ROSE (Repression Of heat Shock gene Expression) which presumably detects temperature changes via reversible RNA folding and unfolding. According to our model, translation initiation signals are masked at low temperatures, whereas melting of the RNA structure at elevated temperatures alleviates repression by allowing ribosome entry too the Shine-Dalgarno region. Computer-based structural predictions og the sensory RNA are supported by mutational analyses and will now be testet by various techniques, including chemical and enzymatic probing experiments. Toe-printing analyses will be used to examine the temperature effect on ribosome binding to the sensory RNA. Completed microbial genome sequences will be searched for other RNA thermometers which will then be studied by a complementary set of in vivo an in vitro techniques. Finally, RNA thermosensors will be exploited for biotechnological purposes by the construction of temperature-controllable expression vectors.

直到最近，直接感知环境信号才被认为是某些 mRNA 的固有特性。感觉 RNA 分子 5`-非翻译区 (5`-UTR) 的构象变化预计将控制下游基因的转录或翻译。我们目前正在研究 RNA 温度计 ROSE（热休克基因表达抑制），它可能通过可逆的 RNA 折叠和展开来检测温度变化。根据我们的模型，翻译起始信号在低温下被掩盖，而在高温下RNA结构的解链通过允许核糖体进入Shine-Dalgarno区域来减轻抑制。基于计算机的感觉 RNA 结构预测得到突变分析的支持，现在将通过各种技术进行测试，包括化学和酶探测实验。脚趾打印分析将用于检查温​​度对核糖体与感觉 RNA 结合的影响。将搜索完整的微生物基因组序列以寻找其他 RNA 温度计，然后通过一套互补的体内和体外技术进行研究。最后，通过构建温度可控的表达载体，RNA 热传感器将被用于生物技术目的。