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.

果蝇细菌颗粒中的mRNA组装 每个物种的开发都依赖于空间组织的使者RNA（mRNA）。确实，甚至是 单个不对称分布的mRNA可以指定细胞的形态，即发育的身体计划 有机体和细胞谱系跨后生动物。 RNA结合蛋白（RBP）通常组织mRNA和 将它们招募到亚细胞结构。但是，mRNA也可以自组织成多MRNA组件 独立于其他细胞成分。在不同物种的健康细胞中发现mRNA组件 并且通常与RNA颗粒，调节的无膜核糖核蛋白（RNP）颗粒有关 mRNA的翻译和稳定性。 mRNA组件也是人类重复发病机理的标志 膨胀障碍，例如肌营养不良症1（MD1），肌萎缩性侧索硬化症（ALS）和 脊椎动物共济失调（SCA）。这些发现表明mRNA组装是mRNA的固有特性 这可能与生物学相关。但是，mRNA的确切机制和潜在的生物学功能 组装尚不清楚。 MRNA已显示通过相分离可以自组装，该过程类似于油和 - 水去混合。在这里，RNA：RNA相互作用，促进分子间（反式）碱基对和 次级（顺式）RNA结构与驱动mRNA组装有关。另外，体外数据 认为是RNA颗粒的核心成分的RNA解旋酶被认为可以溶解RNA：RNA 颗粒中的相互作用。鉴于这些发现，出现了几个关键问题。 mRNA如何 旨在预防RNA的颗粒中的自组装：RNA相互作用？ mRNA组件在 RNA颗粒？反式和顺式RNA的功能是什么：RNA相互作用和RNA解旋酶在 mRNA组件和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 果蝇细菌颗粒中的组装。 mRNA组件倾向于形成为敏锐的小簇 对它们形成的浓度和环境敏感。因此，核心挑战在 研究mRNA组装是必须采用能够在 体内且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 成像方法以分析完整细胞和自然细胞中的mRNA组装 语境。我们旨在发现有机体如何利用mRNA组装来促进的新见解 发育以及mRNA组装的不调节可能导致人类疾病，例如MD1， ALS和SCA。