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.

果蝇胚芽颗粒中的 mRNA 组装 每个物种的发展都严重依赖于空间组织的信使 RNA (mRNA)。确实，即使是一个 单个不对称分布的 mRNA 可以指定细胞的形态、发育中的身体结构 跨后生动物的有机体和细胞谱系。 RNA 结合蛋白 (RBP) 通常组织 mRNA 并 将它们招募到亚细胞结构中。然而，mRNA 也可以自组织成多 mRNA 组件 独立于其他细胞成分。 mRNA 组装体存在于不同物种的健康细胞中 通常与 RNA 颗粒、无膜核糖核蛋白 (RNP) 颗粒相关，这些颗粒调节 mRNA 的翻译和稳定性。 mRNA 组装也是人类重复发病机制的一个标志 扩张障碍，例如强直性肌营养不良 1 (MD1)、肌萎缩侧索硬化症 (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 这可能具有生物学意义。然而，mRNA的确切机制和潜在的生物学功能 组装不清楚。 mRNA 已被证明可以通过相分离进行自组装，这一过程类似于油和- 水分层。在这里，RNA：RNA 相互作用，促进分子间（反式）碱基配对和 二级（顺式）RNA 结构与驱动 mRNA 组装有关。此外，体外数据 表明 RNA 解旋酶（RNA 颗粒的核心成分）被认为可以溶解 RNA:RNA 颗粒中的相互作用。根据这些发现，出现了几个关键问题。 mRNA 是怎样做的 自组装成颗粒以防止 RNA:RNA 相互作用？ mRNA组装的作用是什么 RNA颗粒？反式和顺式 RNA:RNA 相互作用和 RNA 解旋酶的功能是什么 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 果蝇胚芽颗粒中的组装。 mRNA 组装体倾向于形成小簇，这些簇在 对它们形成的浓度和环境敏感。因此，核心挑战是 研究 mRNA 组装的关键在于必须采用能够在 体内并具有高分辨率和灵敏度。我们将使用遗传和定量的超分辨率 成像方法分析完整细胞及其天然细胞内的 mRNA 组装 语境。我们的目标是揭示生物体如何利用 mRNA 组装来促进 mRNA 组装的错误调控如何导致人类疾病，例如 MD1、 ALS 和 SCA。