STRUCTURE DETERMINATION OF AN INTACT BACTERIAL SELF-SPLICING INTRON

完整细菌自剪接内含子的结构测定

• 批准号: 7726211
• 负责人: SCOTT A STROBEL
• 金额: $ 3.11万
• 依托单位: BROOKHAVEN SCIENCE ASSOC-BROOKHAVEN LAB
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-09-18 至 2009-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7726211
• 关键词: Active Sites; Biochemistry; Biological; Biological Process; Catalysis; Catalytic RNA; Chemicals; Class; Communities; Computer Retrieval of Information on Scientific Projects Database; Experimental Designs; Funding; Goals; Grant; Group Structure; Heavy Metals; Heterogeneous Nuclear RNA; Institution; Introns; Ions; Life; Metals; Nature; Object Attachment; Organism; Peptides; Process; Protein Biosynthesis; RNA; RNA Splicing; Reaction; Research; Research Personnel; Resolution; Resources; Ribosomes; Role; Source; Structure; United States National Institutes of Health; Work; base; catalyst; inhibitor/antagonist; insight; interest; mRNA Precursor; metalloenzyme; progenitor

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. There are several classes of naturally occurring catalytic RNAs or ribozymes. The importance of understanding how this class of biocatalysts promotes their reactions became evident with the recent structure determination of the 50S ribosome, which demonstrated that protein synthesis in all living organisms occurs within an RNA active site. Ribozymes are the molecules most likely to be the progenitors of modern biological catalysts and understanding how they promote their reactions provides critical insight into enzymological function. This project focuses on the structure of the group I self-splicing intron, the first class of catalytic RNAs discovered. Structural information is necessary to understand and interpret the biological function of this class of biocatalyst. We are working toward three goals. 1. The primary aim of this project is to determine the high resolution x-ray crystal structure of an intact bacterial group I self-splicing intron. If this aim is achieved, two additional projects will be undertaken: 2. Heavy metal ions and alternative intron inhibitors will be used to structurally investigate how metal ions contribute to group I intron folding and catalysis. 3. The transition state of the phosphotransfer reaction will be structurally explored using stable mimics of the transition state geometry. These studies will provide a structural basis for more than 20 years of biochemistry. It will also provide chemical insights into the nature of RNA catalysis, particularly for an RNA metalloenzyme, an example of which has not yet been structurally analyzed. RNA catalysis is an issue of particular interest given the critical of role of RNA in several natural processes including peptide bond formation and pre-mRNA splicing. The coordinates will be made available to the scientific community for analysis and experimental design.

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目和 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 对于中心来说，它不一定是研究者的机构。 天然存在的催化 RNA 或核酶有几类。 随着最近 50S 核糖体结构的确定，了解此类生物催化剂如何促进其反应的重要性变得显而易见，这表明所有活生物体中的蛋白质合成都发生在 RNA 活性结构中 地点。 核酶是最有可能成为现代生物催化剂祖先的分子，了解它们如何促进反应可以为酶学功能提供重要的见解。 该项目重点研究 I 组自剪接内含子的结构，这是发现的第一类催化 RNA。 结构信息对于理解和解释此类生物催化剂的生物功能是必要的。 我们正在努力实现三个目标。 1. 该项目的主要目的是确定完整细菌 I 组自剪接内含子的高分辨率 X 射线晶体结构。 如果实现这一目标，将实施另外两个项目： 2. 重金属离子和替代内含子抑制剂将用于结构研究金属离子如何促进 I 族内含子折叠和催化。 3. 将使用过渡态几何结构的稳定模拟物对磷酸转移反应的过渡态进行结构探索。 这些研究将为20多年的生物化学提供结构基础。 它还将为 RNA 催化的本质提供化学见解，特别是 RNA 金属酶，其示例尚未进行结构分析。 鉴于 RNA 在肽键形成和前 mRNA 剪接等多种自然过程中的关键作用，RNA 催化是一个特别令人感兴趣的问题。 这些坐标将提供给科学界用于分析和实验设计。