Function and structure of the RNA-processing exosome in the hyperthermophilic archaeon Sulfolobus solfataricus

超嗜热古菌硫磺硫化叶菌 RNA 加工外泌体的功能和结构

• 批准号: 5450117
• 负责人: Professorin Dr. Gabriele Klug
• 金额: --
• 依托单位: Institut für Mikrobiologie und Molekularbiologie
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2005
• 资助国家: 德国
• 起止时间: 2004-12-31 至 2014-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/5450117?language=en
• 关键词: Function; structure; RNA; processing; exosome

While different aspects of RNA processing and degradation are well studied in Eubacteria and Eukarya, little is known about these essential processes in Archaea, the third domain of life. We have isolated a protein complex from the hyperthermophilic archaeon Sulfolobus solfataricus which is a simplified version of the eukaryotic exosome. The eukaryotic exosome is involved in maturation and degradation of RNA, its subunits are essential for cell viability. This project aims to unravel the physiological function of the S. solfataricus exosome and to identify its natural RNA substrates. The in vivo function of the exosome subunits will be studied by gene silencing in S. solfataricus and by complementation of conditional yeast mutants. In addition the biochemical activity of the subunits will be determined in vitro. The exact structure of the eukaryotic exosome could not be solved to date. We like to take advantage of the simpler organization of the archaeal exosome for structural analyses. We will study protein-protein interaction between the individual subunits and reconstitute complexes of different composition for electron microscopic and crystallographic analyses. Our results will build the fundament for future investigations of gene regulation at post-transcriptional level in Archaea, will contribute to our understanding of protein-protein and RNA-protein interactions at high temperatures and will provide interesting aspects on the evolution of RNA processing protein complexes.

虽然RNA加工和降解的不同方面在真细菌和真核生物中得到了很好的研究，但对生命的第三个领域--真核生物中的这些基本过程知之甚少。我们已经从极端嗜热古菌Sulfolobus solfataricus中分离出一种蛋白复合物，它是真核外泌体的简化版本。真核生物外泌体参与RNA的成熟和降解，其亚基对细胞的生存至关重要。本项目旨在揭示S. solfataricus外泌体的研究和鉴定其天然RNA底物。外泌体亚基的体内功能将通过在S. solfataricus和条件酵母突变体的互补。此外，将在体外测定亚基的生物化学活性。真核细胞外泌体的确切结构至今还没有解决。我们喜欢利用古细菌外泌体的简单组织进行结构分析。我们将研究单个亚基之间的蛋白质-蛋白质相互作用，并重组不同组成的复合物进行电子显微镜和晶体学分析。我们的研究结果将为今后在转录后水平上研究拟南芥的基因调控奠定基础，将有助于我们理解蛋白质-蛋白质和RNA-蛋白质在高温下的相互作用，并将为RNA加工蛋白复合物的进化提供有趣的方面。