Aberrant P-bodies accumulation and clearance in yeast and human cells.

酵母和人体细胞中异常 P 体的积累和清除。

• 批准号: 10551880
• 负责人: Nava Segev
• 金额: $ 19.99万
• 依托单位: UNIVERSITY OF ILLINOIS AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-01-18 至 2024-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10551880
• 关键词: Address; Adverse effects; Affect; Autophagocytosis; Behavior; Biological; Biological Models; Brain; Caring; Cell Line; Cell Survival; Cell model; Cell physiology; Cells; Child; Cytoplasmic Granules; Data; Defect; Development; Disease; Economic Burden; Enzymes; Etiology; Family; Fibroblasts; Genes; Goals; Growth; Human; Individual; Intellectual functioning disability; Knowledge; Liquid substance; Lysosomes; Membrane; Messenger RNA; Missense Mutation; Mission; Modeling; Molecular Genetics; Mutation; National Institute of Neurological Disorders and Stroke; Neurites; Neurodevelopmental Disorder; Neurons; Nonsense-Mediated Decay; Organelles; Pathogenicity; Pathway interactions; Patients; Phase; Phenotype; Play; Population; Predisposition; Proteins; RNA; RNA Processing; RNA helicase A; Regulation; Research; Role; Severities; Societies; Stress; Synaptic plasticity; System; Toxic effect; Transfection; Ubiquitin; Variant; Yeasts; cell growth; cell immortalization; cell motility; de novo mutation; early onset; genome sequencing; helicase; human disease; human tissue; insight; mRNA Decay; multicatalytic endopeptidase complex; mutant; novel; novel therapeutic intervention; response; severe intellectual disability; tissue/cell culture; yeast protein

Abstract RNA-protein (RNP) granules are dynamic membrane-less organelles that form during normal growth and in response to stress in a reversible manner. One type of RNP granules is Processing Bodies (P-bodies, or PBs), which contain mRNA together with RNA processing enzymes. Their roles include sequestration of malfunctioning mRNA destined for degradation and storing and silencing of mRNAs when not needed. PBs components and behavior are highly conserved between yeast and human cells. Our understanding of composition and assembly by liquid-phase separation normal PBs has progressed in the last decade. In contrast, our knowledge about involvement of PBs in human disease is scarce as is evidence about the occurrence of aberrant PBs and clearance of normal or aberrant PBS. In this project, we propose to study variants in a PBs component associated with a neurodevelopmental disorder that causes intellectual disability. Currently, there is no cure for such disorders. We hypothesize that the variants cause accumulation of aberrant PBs that, unlike normal PBs, are not reversible. The human variants carry missense mutations in residues identical in the yeast protein. Therefore, we started by modeling the effects of these mutations on RNA processing and accumulation of PBs in yeast. While normal PBs assemble and disassemble according to cellular needs, our preliminary evidence in yeast cells points to accumulation of persistent PBs as the major phenotype of these mutations. We propose to study the effects of these mutations in yeast and in human tissue culture cells, including neuronal cell lines. The phenotypes we propose to assess are on the accumulation and dynamics of persistent PBs, including adverse effects they might have on cell function. In addition, we will explore possible pathways that can clear normal and persistent PBs, such as macro- and micro-autophagy and ubiquitin-associated degradation. For these studies we will use a combination of molecular genetics and cellular approaches. Most approaches for studying RNP granules, granule clearance pathways in yeast and human cells are established in our lab. Achieving the goals of this proposal would provide novel paradigms on existence and behavior of persistent PBs and the role they play in a neurodevelopmental disorder. Moreover, identifying pathways that can clear normal and aberrant PBs would provide novel therapeutic strategies for a neurodevelopmental disorder associated with intellectual disability, in line with the missions of the National Institute of Neurological Disorders and Stroke.

摘要 RNA-蛋白质（RNP）颗粒是在正常过程中形成的动态无膜细胞器， 生长和以可逆的方式响应应力。一种类型的RNP颗粒是 加工体（P体或PB），包含mRNA和RNA加工 内切酶它们的作用包括隔离注定要降解的故障mRNA 以及在不需要时储存和沉默mRNA。PB的组件和行为是 在酵母和人类细胞之间高度保守。我们对构图的理解， 通过液相分离组装常规PB在过去十年中已经取得了进展。与此相反， 我们对PBs参与人类疾病的了解很少， 异常PBS的发生和正常或异常PBS的清除。 在这个项目中，我们建议研究与一种 导致智力残疾的神经发育障碍。目前，没有治愈 这样的紊乱。我们假设这些变异导致异常PB的积累， 与正常的PB不同，是不可逆的。人类变异体在残基中携带错义突变 与酵母蛋白质相同。因此，我们开始模拟这些突变的影响， RNA加工和PBs在酵母中的积累。当正常的PB组装时， 根据细胞的需要进行分解，我们在酵母细胞中的初步证据表明， 持续性PB的积累是这些突变的主要表型。我们建议 研究这些突变在酵母和人类组织培养细胞中的影响，包括 神经元细胞系我们建议评估的表型是在积累和 持续性PB的动力学，包括它们可能对细胞功能产生的不利影响。在 此外，我们还将探索清除正常和持续性PB的可能途径，例如 宏观和微观自噬和泛素相关降解。对于这些研究，我们将 使用分子遗传学和细胞方法的组合。大多数学习方法 本实验室建立了RNP颗粒、酵母和人细胞中的颗粒清除途径。 实现这一建议的目标将提供关于存在和行为的新范式 以及它们在神经发育障碍中的作用。此外，识别 可以清除正常和异常PB的途径将为治疗糖尿病提供新的治疗策略。 一种与智力残疾有关的神经发育障碍，符合 国家神经疾病和中风研究所