Structure and Function of Non-Coding RNA

非编码RNA的结构和功能

• 批准号: 10623993
• 负责人: Joseph Anthony Piccirilli
• 金额: $ 81.31万
• 依托单位: UNIVERSITY OF CHICAGO
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-06-01 至 2028-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10623993
• 关键词: 3-Dimensional; Affinity; Architecture; Area; Binding; Biochemical; Biological Models; Biological Process; Biology; Biophysics; Catalytic RNA; Cell physiology; Chemicals; Code; Communities; Complex; Cryoelectron Microscopy; Crystallization; Crystallography; Data; Development; Disease; Engineering; Fab Immunoglobulins; Genetic; Health; Human; Investigation; Isotopes; Kinetics; Knowledge; Laboratories; Link; Macromolecular Complexes; Mediating; Molecular Chaperones; Molecular Structure; Nucleotides; Play; Proteins; RNA; Reagent; Recombinants; Resolution; Role; Shapes; Specificity; Structure; System; Testing; Time; Transcript; Untranslated RNA; Work; biochemical tools; catalyst; density; macromolecule; novel; novel strategies; nucleotide analog; protein complex; protein function; small molecule; structural biology; synthetic antibodies; tool; transcriptome

PROJECT SUMMARY/ABSTRACT My group develops and uses chemical and biochemical tools to investigate the structure and function of noncoding RNAs (ncRNA) and their complexes with proteins (RNPs) and small molecules (smRNAs). In the context of this project, we will specifically focus on two general areas: 1) engineering fragment antigen binding (Fabs) as chaperones for RNA/RNP/smRNA crystallography, cryoelectron microscopy (cryoEM) and as enabling tools for RNA biology, and 2) elaborating mechanisms of RNA function, especially catalytic RNAs (ribozymes). The terrestrial transcriptome contains vast numbers of structured RNAs within both coding and noncoding transcripts that act alone and in concert with proteins to carry out critically important roles in nearly every facet of cellular function. Understanding how these RNAs mediate biological function in health and disease requires knowledge of their three-dimensional architectures. We will deploy a variety of structural biology, biophysical, biochemical, computational and chemical approaches, including kinetic isotope effect analysis and the use of nucleotides from an artificially expanded genetic system. The data that emerge from our recombinant Fab work will engender a robust platform to generate Fabs that bind structured RNA with high affinity and specificity, establish Fabs as powerful reagents in structural and mechanistic RNA biology and yield new macromolecular structures, which have profound impact on our understanding of the underlying biology and serve as a framework to develop and test functional hypotheses. In addition, our in-depth work on catalytic RNA will define for the first time the relationship between catalytic structure and transition state structure, a missing link that has plagued efforts to engineer macromoleclules with novel catalytic functions. Beyond the conceptual advances that emerge, this work promises to deliver to the RNA community powerful Fab reagents for RNA/RNP structural and functional studies, nucleotide analogs to support novel approaches to define RNA catalytic mechanisms, and strategies to render large RNAs more tractable by increasing the density of information.

项目摘要/摘要 我的小组开发并使用化学和生化工具来研究 非编码RNA（NCRNA）及其与蛋白质（RNP）和小分子（SMRNA）的复合物。在 该项目的背景，我们将专门关注两个一般领域：1）工程片段抗原绑定 （Fabs）作为RNA/RNA/RNP/SMRNA晶体学的伴侣，冷冻电子显微镜（冷冻）和启用 RNA生物学的工具，以及2）阐述RNA功能的机制，尤其是催化RNA（核酶）。 地面转录组包含编码和非编码中的大量结构化RNA 单独起作用并与蛋白质共同发挥作用的成绩单，几乎在每个方面都非常重要 细胞功能。了解这些RNA如何介导健康和疾病中的生物学功能需要 了解他们的三维体系结构。我们将部署各种结构生物学，生物物理， 生化，计算和化学方法，包括动力学同位素效应分析和使用 人为扩展的遗传系统的核苷酸。我们的重组工厂从 将产生一个强大的平台，以生成具有高亲和力和特异性结构的RNA的Fabs， 建立晶圆厂作为结构和机械RNA生物学中的强大试剂，并产生新的大分子分子 结构对我们对潜在生物学的理解产生深远影响并作为框架 开发和检验功能假设。此外，我们对催化RNA的深入研究将定义为第一个 时间催化结构与过渡状态结构之间的关系，这是一个困扰的缺失联系 为大型催化功能设计大分子的努力。超越了出现的概念进步， 这项工作有望将RNA/RNP结构的功能强大的Fab试剂交付给 功能研究，核苷酸类似物，以支持定义RNA催化机制的新方法，并 通过增加信息密度来使大型RNA更具处理性的策略。