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: 10390634
• 负责人: Nava Segev
• 金额: $ 23.99万
• 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 2023-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10390634
• 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; Lead; 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; base; 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.

抽象的 RNA蛋白（RNP）颗粒是在正常情况下形成的无动态膜细胞器 增长和应对压力的反应。一种RNP颗粒是 加工体（P-Bodies或PBS），其中包含mRNA以及RNA处理 酶。它们的作用包括隔断发病的mRNA注定降解的mRNA 以及在不需要的情况下存储和沉默mRNA。 PBS组件和行为是 酵母和人类细胞之间高度保守。我们对构图和 在过去的十年中，通过液相分离的组装正常PBS已进展。相比之下， 我们对PBS参与人类疾病的参与的了解很少，这是关于 异常PBS的发生以及正常或异常PBS的清除。 在这个项目中，我们建议在与A相关的PBS组件中研究变体 导致智力障碍的神经发育障碍。目前，无法治愈 这样的疾病。我们假设变体会导致异常PB的积累，从而 与普通PBS不同，不可逆。人类变种带有残基的错义突变 在酵母蛋白中相同。因此，我们首先要建模这些突变的效果 关于酵母中PBS的RNA处理和积累。而正常的PBS组装和 根据细胞需求拆卸，我们在酵母细胞中的初步证据表明 持久性PBS作为这些突变的主要表型的积累。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关于积累和 持续性PBS的动力学，包括它们可能对细胞功能产生的不良影响。在 此外，我们将探索可能清除正常和持久PBS的可能途径，例如 宏观和微自噬和泛素相关的降解。对于这些研究，我们将 结合分子遗传学和细胞方法。大多数学习方法 在我们的实验室中建立了RNP颗粒，酵母中的颗粒清除途径和人类细胞。 实现该提议的目标将为存在和行为提供新颖的范式 持续的PBS及其在神经发育障碍中的作用。而且，识别 可以清除正常和异常PBS的途径将为您提供新颖的治疗策略 与智力障碍相关的神经发育障碍，与任务一致 国家神经系统疾病和中风研究所。