Role of structural dynamics in RNA regulation

结构动力学在 RNA 调控中的作用

基本信息

批准号：
10685422
负责人：
Anthony McDowell Mustoe
金额：
$ 40万
依托单位：
BAYLOR COLLEGE OF MEDICINE
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-09-01 至 2027-06-30
项目状态：
未结题

项目摘要

Project Summary Messenger RNAs (mRNAs) encode complex regulatory instructions in their 3' untranslated regions (3'UTRs) that prescribe context-dependent control of translation and decay. Disruption of mRNA post-transcriptional regulation by mutational lesions or shortening of 3'UTRs has been linked to diverse human diseases. RNA molecules fold back on themselves into base-paired secondary structures and occasionally higher-order tertiary structures, and sequence-encoded structures are hypothesized to play key roles in directing 3'UTR regulation. For example, RNA structures can provide specific binding sites for RNA binding proteins and may tune the accessibility of microRNA target sites. RNA structures may also function as molecular switches, with protein or microRNA binding triggering specific 3'UTR structural changes that alter the activity of distal regulatory sites. However, little remains known about 3'UTR structures and their role in post-transcriptional regulation. Existing data indicate that 3'UTRs have heterogenous and dynamic structures, which makes them challenging to study by traditional chemical probing or biophysical methods. My research group has recently developed a new single-molecule chemical probing technology (DANCE-MaP) that makes it possible to accurately define RNA structural ensembles consisting of 2-3 coexisting states in living cells. We will pursue two areas of research that exploit and build on the DANCE-MaP technology: (1) We will apply DANCE- MaP to define 3'UTR folding landscapes and mechanisms of combinatorial regulation by RNA binding proteins and microRNAs. (2) We will develop next-generation technologies that can resolve structurally heterogenous RNAs with greater resolution and provide model-free, physical interpretation of chemical probing data. This work will provide an improved understanding of mRNA regulatory mechanisms, helping advance efforts to therapeutically target 3'UTRs to modulate gene expression, and contribute new structure determination technologies that will be of broad utility to the RNA biology field.

项目概要信使 RNA (mRNA) 在其 3' 非翻译区编码复杂的调节指令区域（3'UTR）规定了翻译和衰变的上下文相关控制。破坏突变损伤或 3'UTR 缩短对 mRNA 转录后调控的影响与多种人类疾病有关。 RNA 分子自行折叠成碱基配对二级结构和偶尔更高阶的三级结构，以及序列编码假设结构在指导 3'UTR 调节中发挥关键作用。例如，RNA 结构可以为 RNA 结合蛋白提供特异性结合位点，并可以调节 microRNA 靶位点的可及性。 RNA结构也可以充当分子开关，与蛋白质或 microRNA 结合触发特定的 3'UTR 结构变化，从而改变远端调节位点的活性。然而，人们对 3'UTR 结构及其作用知之甚少。转录后调控中的作用。现有数据表明 3'UTR 具有异质性和动态结构，这使得通过传统化学探测或生物物理方法。我的研究小组最近开发了一种新的单分子化学探测技术 (DANCE-MaP) 可准确定义 RNA 由活细胞中 2-3 个共存状态组成的结构整体。我们将致力于两个领域开发和建立 DANCE-Map 技术的研究：(1) 我们将应用 DANCE- MaP 定义 3'UTR 折叠景观和 RNA 组合调节机制结合蛋白和 microRNA。 (2)我们将开发下一代技术以更高的分辨率解析结构异质 RNA，并提供无模型、物理化学探测数据的解释。这项工作将加深对 mRNA 调控机制，有助于推进以 3'UTR 为治疗目标的努力调节基因表达，并贡献新的结构测定技术在 RNA 生物学领域具有广泛的用途。