Global Structures of RNA

RNA 的整体结构

• 批准号: 7228626
• 负责人: David P MILLAR
• 金额: $ 27.15万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-04-01 至 2009-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7228626
• 关键词: 7SL RNA; Address; Adopted; Biological Process; Catalytic RNA; Complex; DNA Sequence Rearrangement; Fluorescence; Fluorescence Resonance Energy Transfer; Goals; Kinetics; Lead; Learning; Ligands; Mediating; Metal Ion Binding; Methods; Modeling; Molecular Chaperones; Monitor; Pathway interactions; Process; Protein Biosynthesis; Proteins; RNA; RNA Folding; Range; Role; Signal Recognition Particle; Site; Structure; System; Testing; Therapeutic; base; cancer therapy; cofactor; conformational conversion; design; gene therapy; hairpin ribozyme; improved; particle; single molecule; structural biology; three dimensional structure; tool

DESCRIPTION (provided by applicant): To achieve their diverse range of biological functions, RNA molecules must fold into specific tertiary structures that create catalytic centers or ligand recognition sites. Despite the recent emergence of a wealth of information on the three-dimensional structures of RNA molecules and RNA-protein complexes, culminating in structures of ribosomal particles, the mechanisms by which RNA adopts these structures are poorly understood. The broad, long-term objective of this proposal is to understand the mechanism of RNA folding. Fundamental features of RNA folding include conformational search, metal ion binding and productive intermediates. Additionally, most large RNA molecules require protein cofactors in order to fold correctly and efficiently. Hence, we will continue to analyze tertiary structure formation in the hairpin ribozyme, an autonomously folding RNA, and we will also undertake new studies of the folding of the 7SL RNA from the mammalian signal recognition particle (SRP), which folds in the presence of multiple protein chaperones. By studying both systems, we hope to learn about the general principles of RNA folding. Moreover, the results will facilitate the design of improved ribozymes for therapeutic applications in gene therapy and, eventually, may lead to new cancer therapies based on arrest of protein synthesis by the SRP machinery. The specific aims are: (1) Elucidate the role of loop rearrangements and dynamic loop structure in tertiary structure formation in the hairpin ribozyme. (2) Test the proposed conformational search model for tertiary structure formation in the hairpin ribozyme and identify determinants of the search. (3) Monitor RNA folding transitions during assembly of the Alu domain of the signal recognition particle. (4) Elucidate the mechanism of the initial assembly of the S-domain of the signal recognition particle and characterize associated RNA folding transitions. To address these goals, we will apply a range of ensemble and single-molecule fluorescence methods to dissect the RNA folding pathways and to monitor dynamic conformational transitions during folding. Additionally, the experimental methods developed during this project will provide new tools for studying the conformational dynamics and folding of RNA and will be broadly applicable.

描述（由申请人提供）：为了实现其多种生物学功能，RNA分子必须折叠成特定的三级结构，以产生催化中心或配体识别位点。尽管最近出现了大量的信息，RNA分子和RNA-蛋白质复合物的三维结构，最终在核糖体颗粒的结构，RNA采用这些结构的机制知之甚少。这项计划的长期目标是了解RNA折叠的机制。RNA折叠的基本特征包括构象搜索、金属离子结合和生产性中间体。此外，大多数大RNA分子需要蛋白质辅因子才能正确有效地折叠。因此，我们将继续分析发夹状核酶（一种自主折叠的RNA）中三级结构的形成，并且我们还将对哺乳动物信号识别颗粒（SRP）中7SL RNA的折叠进行新的研究，该颗粒在多个蛋白伴侣存在下折叠。通过研究这两个系统，我们希望了解RNA折叠的一般原理。此外，这些结果将有助于设计改进的核酶在基因治疗中的治疗应用，并最终可能导致新的癌症治疗的基础上逮捕的蛋白质合成的SRP机制。具体目标是：（1）阐明发夹状核酶中环状结构重排和动态环状结构在三级结构形成中的作用。(2)测试所提出的发夹状核酶三级结构形成的构象搜索模型，并确定搜索的决定因素。(3)在信号识别颗粒的Alu结构域组装期间监测RNA折叠转变。(4)阐明信号识别颗粒S结构域初始组装的机制，并表征相关的RNA折叠转变。为了实现这些目标，我们将应用一系列的合奏和单分子荧光方法来剖析RNA折叠途径，并监测折叠过程中的动态构象转变。此外，在本项目中开发的实验方法将为研究RNA的构象动力学和折叠提供新的工具，并将具有广泛的适用性。