mRNA assembly in Drosophila germ granules

果蝇胚芽颗粒中的 mRNA 组装

• 批准号: 10275109
• 负责人: Tatjana Trcek
• 金额: $ 40.94万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-08-12 至 2026-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10275109
• 关键词: Acute; Amyotrophic Lateral Sclerosis; Automobile Driving; Base Pairing; Biological; Biological Process; Cell Lineage; Cells; Cellular Morphology; Cytoplasmic Granules; Data; Development; Disease; Drosophila genus; Environment; Genetic; Genetic Transcription; Germ; Goals; Human; In Vitro; Light; Messenger RNA; Methods; Myotonic Dystrophy; Oils; Organism; Pathogenesis; Phase; Process; Property; RNA; RNA Helicase; RNA-Binding Proteins; Reaction; Resolution; Ribonucleoproteins; Role; Specific qualifier value; Spinocerebellar Ataxias; Structure; Subcellular structure; Transcript; Translations; Water; design; human disease; imaging approach; in vivo; insight; particle; prevent; recruit

mRNA ASSEMBLY IN DROSOPHILA GERM GRANULES Development of every species relies critically on spatially organized messenger RNAs (mRNAs). Indeed, even a single asymmetrically distributed mRNA can specify the morphology of cells, the body plan of a developing organism and cell lineages across metazoans. RNA binding proteins (RBPs) typically organize mRNAs and recruit them to subcellular structures. However, mRNAs can also self-organize into multi-mRNA assemblies independently of other cellular components. mRNA assemblies are found in healthy cells in different species and are often associated with RNA granules, membraneless ribonucleoprotein (RNP) particles that regulate translation and stability of mRNAs. mRNA assemblies are also a hallmark of pathogenesis in human repeat expansion disorders such as myotonic dystrophy 1 (MD1), amyotrophic lateral sclerosis (ALS) and spinocerebellar ataxia (SCA). These findings indicate that mRNA assembly is an inherent property of an mRNA that could be biologically relevant. However, the exact mechanism and potential biological functions of mRNA assembly is unclear. mRNAs have been shown to self-assemble by phase separation, a process akin to oil-and- water de-mixing. Here, RNA:RNA interactions, which promote intermolecular (trans) base-pairing and secondary (cis) RNA structures have been implicated in driving mRNA assembly. In addition, in vitro data suggest that RNA helicases, which are core constituents of RNA granules are thought to dissolve RNA:RNA interactions in granules. In light of these findings, several critical questions arise. How do mRNAs self-assemble in granules designed to prevent RNA:RNA interactions? What is the role of mRNA assemblies in RNA granules? What is the function of trans and cis RNA:RNA interactions and RNA helicases in the formation and function of mRNA assemblies and RNA granules? The five-year goal of this study is to answer these questions and determine the mechanism and biological function of mRNA assembly in Drosophila germ granules. mRNA assemblies tend to form as small clusters that are acutely sensitive to the concentration and the environment in which they form. Thus, the central challenge in studying mRNA assembly is that methods must be employed that enable examination of this process in vivo and with high resolution and sensitivity. We will use genetic and quantitative, super-resolution imaging approaches to analyze mRNA assembly in intact cells and within their natural cellular context. We aim to uncover new insight into how organisms harness mRNA assemblies to promote development and how misregulation of mRNA assembly could contribute to human diseases, such as MD1, ALS and SCA.

果蝇生殖颗粒的mRNA组装 每个物种的发展都依赖于空间组织的信使RNA（mRNA）。事实上， 单个不对称分布的mRNA可以指定细胞的形态，发育中的细胞的体平面， 有机体和细胞谱系的进化。RNA结合蛋白（RBP）通常组织mRNA， 将它们招募到亚细胞结构中。然而，mRNA也可以自组织成多mRNA组装体 独立于其他细胞成分。在不同物种的健康细胞中发现了mRNA组装 并且通常与RNA颗粒、调节细胞凋亡的无膜核糖核蛋白（RNP）颗粒相关。 mRNA的翻译和稳定性。mRNA组装也是人类重复序列中发病机制的标志。 扩张性疾病，例如强直性肌营养不良1（MD 1）、肌萎缩性侧索硬化（ALS）和 脊髓小脑性共济失调（SCA）。这些发现表明，mRNA组装是mRNA的固有特性， 可能与生物学相关的信息然而，mRNA的确切机制和潜在的生物学功能 组装不清楚。mRNA已经被证明通过相分离自组装，这一过程类似于油- 水分层。在这里，RNA：RNA相互作用，促进分子间（反式）碱基配对， 二级（顺式）RNA结构与驱动mRNA装配有关。此外，体外数据 提示RNA解旋酶是RNA颗粒核心成分，被认为溶解RNA：RNA 颗粒间的相互作用根据这些调查结果，出现了几个关键问题。mRNA如何 自组装成颗粒来阻止RNA：RNA相互作用？mRNA组装体在细胞凋亡中的作用是什么？ RNA颗粒？什么是反式和顺式RNA的功能：RNA相互作用和RNA解旋酶在 mRNA组装体和RNA颗粒的形成和功能？这项研究的五年目标是 回答这些问题，并确定mRNA的机制和生物学功能 果蝇生殖颗粒的组装。mRNA组装倾向于形成小簇， 对它们形成的浓度和环境敏感。因此， 研究mRNA组装的关键是必须采用能够检查这一过程的方法， 具有高分辨率和高灵敏度。我们将使用遗传和定量的超分辨率 成像方法来分析完整细胞及其天然细胞内的mRNA组装 上下文我们的目标是揭示生物体如何利用mRNA组装来促进 发展以及mRNA组装的失调如何导致人类疾病，如MD 1， ALS和SCA。