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.

项目摘要 这项研究的总体目标是增加我们对纤毛的理解，如何通过分子解剖 分子水平上睫状运动的机理和调节。纤毛和鞭毛是保守的和无处不在的 基于微管的细胞器，在细胞运动，流体传输，感觉，细胞信号和 开发是许多真核细胞的生存和正常功能的关键过程， 组织。在人类中，纤毛的运动和组装缺陷负责许多先天性 疾病，例如原发性睫状运动障碍，慢性呼吸系统疾病，生育能力受损，脑发育 缺陷，先天性心脏病和左右轴的随机化。纤毛运动是由 由多种同工型组成的数千种动力蛋白分子的协调活性。我们以前关于 野生型和突变的纤毛，并积极击败纤毛，为功能组织打开了一个新窗口 西里亚母亲。但是，关于监管信号如何变化的长期基本问题仍然存在 Dynein在分子水平上的活性，不同调节络合物在纤毛中的作用是什么 运动性，以及如何在空间和临时协调的方式上产生振荡的典型节拍 纤毛。该提案以出版和初步的新数据的强烈前提为基础，直接解决 这些关键差距通过针对（AIM 1）揭示机制的三个特定目标 Dynein的作用受调节以发起和传播睫状波，（AIM 2）确定动力蛋白的模式 产生不同睫状波形的活动，（目标3）表征组装的睫状成分 只有在特定的双打中询问它们固有的不对称分布是否有助于产生睫毛跳动。 我们使用现代方法的强大而创新的组合，包括冷冻电子层析成像 图像突变纤毛和具有分子分辨率，遗传学和蛋白质组学的标记蛋白质，这是一种替代模型 研究纤毛的有机体，以及一种比以前的最先进的“切割”技术，可以看待“内部更深”的细胞 可能的。我们希望我们的合并研究将提供重要的新概念和机械洞察力 进入睫状运动和调节，这也将影响我们对睫毛疾病的理解。