NMR STUDIES OF RNAS AND RNA-PROTEIN INTERACTIONS

RNAS 和 RNA-蛋白质相互作用的 NMR 研究

• 批准号: 2634779
• 负责人: EDWARD P NIKONOWICZ
• 金额: $ 10.47万
• 依托单位: RICE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1995
• 资助国家: 美国
• 起止时间: 1995-01-01 至 1999-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2634779
• 关键词: RNA binding protein; acidity /alkalinity; bacterial virus; chemical binding; conformation; dimer; intermolecular interaction; molecular dynamics; mutant; nuclear magnetic resonance spectroscopy; nucleic acid sequence; nucleic acid structure; protein structure function; solutions; structural biology; virus RNA; virus protein

RNA-protein interactions are critical to many cellular processes including the regulation of gene expression. A great deal of genetic and biochemical data exists for several RNA-protein systems; however, the structural principles that govern RNA specific recognition by RNA binding proteins are poorly understood. The bacteriophage R17 contains one of the most well characterized RNA-protein interactions. The coat protein of R17 binds tightly and specifically to an RNA hairpin containing the translation start site of the viral replicase gene. This study will focus on the application of hetero-nuclear magnetic resonance (NMR) spectroscopy to probe the structural elements that give rise to this interaction. Structural data will be generated for the RNA hairpins in the absence and presence of R17 coat protein. We will also investigate the structure and-dynamics of loop and stem variants that show different degrees of protein binding affinity. Because RNA molecules have the ability to fold to form unique secondary and tertiary structures, RNA binding proteins are forced to distinguish between a diverse array of three dimensional shapes -- each possessing a precise electrostatic arrangement The R17 system represents a convenient model that will allow us to identify a few of the structural components employed by RNA molecules to confer specificity to RNA-protein interactions. The principles governing molecular recognition in the R17 system should be relevant to RNA-protein interactions in eukaryotic cells. The principles discovered here may be used in the preparation of RNA or protein ligands designed to perform a specific task such as site specific cleavage of a nucleic acid fragment in vivo or in vitro.

RNA-蛋白质相互作用对许多细胞过程至关重要 包括基因表达的调节。大量的基因和 存在几种RNA-蛋白质系统的生物化学数据;然而， 通过RNA结合控制RNA特异性识别的结构原理 对蛋白质的了解很少。噬菌体R17含有一种 最好的表征RNA-蛋白质相互作用。R17的外壳蛋白 紧密特异性地结合到含有 病毒复制酶基因的翻译起始位点。本研究将重点 异源核磁共振（NMR）的应用 光谱来探测产生这种现象的结构元素， 互动RNA发夹的结构数据将在 不存在和存在R17外壳蛋白。我们亦会研究 环和茎变体的结构和动力学表现出不同的 蛋白质结合亲和力的程度。 因为RNA分子有能力折叠形成独特的二级结构， 和三级结构，RNA结合蛋白被迫区分 在一系列不同的三维形状之间--每个形状都有 R17系统代表了一种 方便的模型，使我们能够识别一些结构， RNA分子用来赋予RNA-蛋白质特异性的组分 交互. R17中的分子识别原理 系统应该与真核生物中RNA-蛋白质相互作用相关 细胞这里发现的原理可用于制备 设计用于执行特定任务的RNA或蛋白质配体，例如位点 在体内或体外特异性切割核酸片段。