Discovery and Function of Higher-Order RNA Structure

高阶RNA结构的发现和功能

• 批准号: 9754843
• 负责人: Kevin M Weeks
• 金额: $ 58.36万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-06-09 至 2022-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9754843
• 关键词: Area; Automobile Driving; Base Pairing; Biological; Biological Process; Biology; Cells; Chemistry; Dengue; Ebola virus; Elements; Emerging Communicable Diseases; Encapsulated; Genome; Ligands; Messenger RNA; Methods; Pathogenicity; Proteins; RNA; RNA Viruses; RNA analysis; Role; Structural Protein; Structure; Technology; Translating; Untranslated RNA; Viral; Vision; Work; Zika Virus; base; chikungunya; drug development; genetic regulatory protein; mammalian genome; new technology; novel; novel therapeutics; pathogenic virus; small molecule; viral RNA

SUMMARY RNA functions as the central conduit of information exchange in all cells, a role encapsulated in two critical observations. First, a large fraction of emerging infectious diseases are caused by RNA viruses including Ebola, Chikungunya, Zika, and Dengue. Second, a much larger fraction of the mammalian genome is transcribed into diverse kinds of non-coding RNAs (~70%) than is translated into protein (1- 2%). The functions of messenger, non-coding, and viral RNAs are governed by the linear sequence, base-paired secondary structure, higher-order tertiary structure, and quaternary interactions involving proteins and small molecules. Overall, our understanding of the number and complexity of RNA structures and how RNA structure drives diverse biological functions is very limited. Most methods developed to date for analyzing RNA structure in high-throughput ways do not measure structure in a definitive and accurate way, making it difficult to define broad principles for interrelationships between RNA structure and function. We seek to understand the fundamental roles of RNA structure in all areas of biology by pursuing a two-pronged approach involving (1) inventing, developing, and rigorously validating highly accurate chemistry-based technologies for discovery of novel RNA structures and the networks of interactions between RNAs and proteins and then (2) applying these technologies to problems of broad importance. Here we propose to interrogate the structures and interaction partners of the pathogenic Dengue RNA virus and the Xist long non-coding RNA. Throughout this work, we will focus on in-cell analysis of native viral and endogenous RNAs. This work is expected to have long-term impact for three broad reasons. First, RNA elements with higher-order folds and extensive protein networks are likely to be harbingers of function. Second, there are likely to be structural folds that are different from the relatively limited classes of structures that have been analyzed to date. Third, RNA elements with higher-order folds also contain clefts and crevices that are ideal targets for small- molecule ligands – and novel drugs – that modulate biological function by targeting RNA.

概括 RNA 的功能是所有细胞中信息交换的中心管道，这一作用包含在两个方面： 批判性观察。首先，很大一部分新发传染病是由RNA病毒引起的 包括埃博拉、基孔肯雅热、寨卡和登革热。其次，哺乳动物的大部分 基因组转录成多种非编码 RNA (~70%)，而不是翻译成蛋白质 (1- 2%）。信使、非编码和病毒 RNA 的功能由线性序列控制， 碱基配对的二级结构、高阶三级结构和四级相互作用涉及 蛋白质和小分子。总体而言，我们对 RNA 数量和复杂性的了解 RNA结构以及RNA结构如何驱动不同的生物功能是非常有限的。大多数方法 迄今为止开发的用于以高通量方式分析 RNA 结构的方法不能测量结构 明确且准确的方式，使得很难定义之间相互关系的广泛原则 RNA结构和功能。我们寻求了解 RNA 结构在所有领域的基本作用 通过采取双管齐下的方法，包括（1）发明、开发和严格地研究生物学 验证基于化学的高精度技术，以发现新颖的 RNA 结构和 RNA 和蛋白质之间的相互作用网络，然后 (2) 将这些技术应用到 具有广泛重要性的问题。在这里我们建议询问结构和互动伙伴 致病性登革热 RNA 病毒和 Xist 长非编码 RNA。在整个工作中，我们将 专注于天然病毒和内源 RNA 的细胞内分析。这项工作预计将长期有效 影响主要有以下三个原因。首先，具有高阶折叠和广泛蛋白质的RNA元件 网络很可能是功能的先兆。其次，可能存在结构性褶皱 与迄今为止分析的相对有限的结构类别不同。三、RNA 具有高阶折叠的元素还包含裂缝和裂缝，它们是小尺寸的理想目标 通过靶向 RNA 来调节生物功能的分子配体和新型药物。