Dissecting alternate modes and regulation of ciliary motility

剖析纤毛运动的交替模式和调节

• 批准号: 10369615
• 负责人: Daniela Nicastro
• 金额: $ 45.79万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-09-28 至 2024-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10369615
• 关键词: Address; Animal Model; Biological Models; Brain; Cell division; Cells; Chlamydomonas; Cilia; Classification; Complex; Congenital Disorders; Cryo-electron tomography; Cytoskeleton; Data; Defect; Development; Disease; Dynein ATPase; Eels; Esthesia; Eukaryotic Cell; Fertility; Flagella; Freezing; Funding; Generations; Genetic; Genetic Diseases; Goals; Handedness; Human; Ice; Image; Impairment; Infertility; Ions; Label; Left; Life; Link; Liquid substance; Location; Methods; Microtubules; Modeling; Modernization; Molecular; Molecular Conformation; Morphogenesis; Motor; Movement; Organ; Organelles; Pattern; Play; Primary Ciliary Dyskinesias; Process; Protein Isoforms; Proteins; Proteome; Proteomics; Publishing; Randomized; Regulation; Research; Resolution; Role; S-nitro-N-acetylpenicillamine; Sea Urchins; Side; Signal Transduction; Slice; Sperm Head; Structural defect; System; Techniques; Therapeutic Intervention; Thick; Thinness; Tissues; Work; base; cell behavior; cell motility; chronic respiratory disease; cilium motility; congenital heart disorder; innovation; insight; mutant; protein structure; sperm cell; therapeutic development; three dimensional structure; trafficking

PROJECT SUMMARY The overall goal of this research is to increase our understanding of how cilia beat by dissecting the molecular mechanisms and regulation of ciliary motility on a molecular level. Cilia and flagella are conserved and ubiquitous microtubule-based organelles with important roles in cell locomotion, fluid transport, sensation, cell signaling and development, which are critical processes for the survival and proper function of many eukaryotic cells and tissues. In humans, defects in the motility and assembly of cilia are responsible for numerous congenital diseases, such as primary ciliary dyskinesia, chronic respiratory disease, impaired fertility, brain developmental defects, congenital heart disease and randomization of the left-right body axis. Cilia motility is driven by the coordinated activities of thousands of dynein molecules, comprised of multiple isoforms. Our previous studies of wild type and mutant cilia, and actively beating cilia have opened a new window into the functional organization of motile cilia. However, long-standing fundamental questions remain about how regulatory signals change dynein’s activity on a molecular level, what are the roles of the different regulatory complexes during ciliary motility, and how dyneins are spatially and temporally coordinated to generate the oscillatory beating typical for cilia. Building on a strong premise of both published and preliminary new data, this proposal directly addresses these critical gaps through three specific aims that are directed at (Aim 1) revealing mechanisms by which dynein’s action is regulated to initiate and propagate ciliary waves, (Aim 2) determine the patterns of dynein activity that generate different ciliary waveforms, and (Aim 3) characterizing ciliary components that assemble only on specific doublets to ask if their inherently asymmetric distribution contributes to producing ciliary beating. We use a powerful and innovative combination of modern approaches that include cryo-electron tomography to image mutant cilia and tagged proteins with molecular resolution, genetics and proteomics, an alternate model organism to study cilia, and a state-of-the-art “cutting” technique to look “deeper inside” cells than previously possible. We expect that our combined studies will provide important new conceptual and mechanistic insights into ciliary motility and regulation, which will also impact our understanding of ciliary diseases.

项目概要 这项研究的总体目标是通过剖析纤毛的分子结构来增加我们对纤毛如何跳动的理解。 纤毛运动在分子水平上的机制和调节是保守且普遍存在的。 基于微管的细胞器，在细胞运动、液体运输、感觉、细胞信号传导和 发育，这是许多真核细胞生存和正常功能的关键过程 在人类组织中，纤毛的运动和组装缺陷是导致许多先天性缺陷的原因。 疾病，如原发性纤毛运动障碍、慢性呼吸道疾病、生育能力受损、大脑发育障碍 缺陷、先天性心脏病和左右身体轴的随机性是由纤毛驱动的。 数千个动力蛋白分子的协调活动，由多​​种亚型组成。 野生型和突变型纤毛，以及主动击败纤毛，打开了了解功能组织的新窗口 然而，关于调节信号如何变化的长期存在的基本问题仍然存在。 动力蛋白在分子水平上的活性，不同调节复合物在纤毛过程中的作用是什么 运动性，以及动力蛋白如何在空间和时间上协调以产生典型的振荡跳动 该提案以已发布的和初步的新数据为坚实基础，直接解决了这一问题。 通过三个具体目标（目标 1）揭示这些关键差距 调节动力蛋白的作用以启动和传播睫状波，（目标 2）确定动力蛋白的模式 产生不同纤毛波形的活动，以及（目标 3）表征组装的纤毛成分 仅在特定的双峰上询问其固有的不对称分布是否有助于产生纤毛跳动。 我们使用现代方法的强大和创新组合，包括冷冻电子断层扫描 使用分子分辨率、遗传学和蛋白质组学（另一种模型）对突变纤毛和标记蛋白质进行成像 研究纤毛的有机体，以及比以前更深入地观察细胞“内部”的最先进的“切割”技术 我们期望我们的综合研究将提供重要的新概念和机制见解。 纤毛运动和调节，这也将影响我们对纤毛疾病的理解。