Mechanism and Regulation of Axonemal Dynein Arm Assembly in Motile Ciliated Epithelial Cells

运动纤毛上皮细胞轴丝动力蛋白臂组装的机制和调控

• 批准号: 10930194
• 负责人: ZHAOXIA SUN
• 金额: $ 57.65万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-23 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10930194
• 关键词: Acceleration; Amino Acids; Biochemical; Biological Assay; Biological Process; Biology; Candidate Disease Gene; Cell physiology; Cells; Cellular Membrane; Cellular biology; Characteristics; Cilia; Crowding; Cytoplasm; Cytosol; Data; Defect; Disease; Dynein ATPase; Environment; Epithelial Cells; Etiology; Extracellular Fluid; Fertility; Foundations; Future; Genes; Genetic; Genetic Diseases; Goals; Health; Human; Image; Infection; Intervention; Investigation; Knock-in; Left; Light; Liquid substance; LoxP-flanked allele; Lung diseases; Macromolecular Complexes; Mediating; Membrane; Messenger RNA; Microtubules; Modeling; Molecular; Molecular Chaperones; Motor; Mucous body substance; Mus; N-terminal; Names; Organism; Patients; Periodicity; Phase; Physiological; Primary Ciliary Dyskinesias; Proteins; Publishing; RNA; Recurrence; Regulation; Reproduction; Research; Respiratory Tract Infections; Ribosomes; Role; Shapes; Skin; System; Testing; Trachea; Translating; Translations; Xenopus; Zebrafish; appendage; arm; cell motility; ciliopathy; cilium biogenesis; cilium motility; fluid flow; insight; mouse genetics; novel; prevent; pulmonary function decline; symptom management; tool

Motile cilia beat rhythmically to propel cell movement or drive extracellular fluid flow. The functional importance of cilia motility in human health is highlighted by primary ciliary dyskinesia (PCD), a genetic disease caused by cilia motility defects. Patients with PCD display left-right asymmetry defect, reduced fertility and progressive lung disease. Currently there is no specific therapy for PCD and management of symptoms has been the main approach. The dynein arms that power cilia motility comprise multiple components that are pre-assembled in the cytosol and many genes associated with PCD encode components of these dynein arms. Intriguingly, however, a separate group includes proteins that reside in the cytosol and appear to be involved in the assembly of dynein arm subunits and they are called dynein arm assembly factors (DNAAFs). Interestingly, multiple DNAAFs are localized in droplet shaped cytosolic foci. However, the precise function of these foci and the precise molecular function of most DNAAFs remain poorly understood. In addition to the assembly of protein components, how mRNAs and the translation machinery are coordinated to supply dynein arm components stoichiometrically is also largely unknown. Based on extensive preliminary and published data, our central hypothesis is that the co- chaperone proteins Pontin and Reptin are core components of a novel membraneless cytosolic assemblage distinct from droplets formed through LLPS; they function to coordinate the translation, folding and assembly of axonemal dynein arm components and prevent aggregate formation. In this project, we will combine zebrafish genetics, mouse genetics and cultured tracheal cells to test our central hypothesis. We propose two specific aims to achieve this goal. In the first aim, we will dissect the molecular and cellular function of Pontin-Reptin foci in discrete steps in dynein arm assembly. In the second aim, we will characterize Pontin-Reptin foci and investigate the mechanism of foci formation. Successful completion of this project will not only provide a molecular framework for dynein arm assembly and the etiology of PCD, but also lay the foundation for future investigation into the regulation, and possible intervention, of dynein arm assembly and cilia motility under diverse physiological and disease conditions.

运动纤毛有节奏地跳动以推动细胞运动或驱动细胞外液流动。功能重要性 原发性纤毛运动障碍（PCD）是一种遗传性疾病， 纤毛运动缺陷PCD患者表现为左右不对称性缺陷、生育能力下降和肺进展 疾病目前，PCD尚无特异性治疗方法，症状管理是主要的治疗方法。 approach.为纤毛运动提供动力的动力蛋白臂包括多个组件，这些组件预先组装在纤毛中。 细胞质和许多与PCD相关的基因编码这些动力蛋白臂的组分。然而，有趣的是， 另一组蛋白质包括存在于胞质溶胶中的蛋白质，它们似乎参与了动力蛋白的组装 它们被称为动力蛋白臂组装因子（DNAAF）。有趣的是，多个DNAAF 局限于液滴状胞质病灶。然而，这些焦点的精确功能和精确的分子 大多数DNAAF的功能仍然知之甚少。除了蛋白质成分的组装， mRNA和翻译机制协调以化学计量地提供动力蛋白臂组分， 大部分也是未知的。基于广泛的初步和已发表的数据，我们的中心假设是，共同- 伴侣蛋白Pontin和Reptin是一种新的无膜细胞质组合的核心成分 与通过LLPS形成的液滴不同;它们的功能是协调 轴丝动力蛋白臂组件和防止聚集体的形成。在这个项目中，我们将联合收割机 遗传学，小鼠遗传学和培养的气管细胞来测试我们的中心假设。我们提出两个具体的 旨在实现这一目标。在第一个目标中，我们将剖析脑桥-Reptin灶的分子和细胞功能 在动力蛋白臂组装中以离散的步骤进行。在第二个目标中，我们将描述脑桥-雷普廷病灶， 探讨病灶形成的机制。该项目的成功完成不仅将提供一个 为动力蛋白臂组装分子框架和PCD的病因学研究，也为今后的研究奠定基础 研究动力蛋白臂组装和纤毛运动的调节和可能的干预， 不同的生理和疾病条件。