Mechanisms underlying centriole morphogenesis

中心粒形态发生的机制

• 批准号: 10370243
• 负责人: Dhivya Kumar
• 金额: $ 10万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-02-01 至 2024-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10370243
• 关键词: Address; Advisory Committees; Age; Architecture; Biochemical; Biological Assay; Biophysics; Brain Diseases; Cell Cycle; Cell physiology; Cells; Centrioles; Centrosome; Cilia; Complex; Cytoplasmic Granules; Daughter; Defect; Development; Digit structure; Disease; Distal; Erinaceidae; Etiology; Face; Foundations; Functional disorder; Goals; Heart; Human; Human Development; Image; Imaging Techniques; In Vitro; Inherited; Joubert syndrome; Length; Lighting; Limb Development; Mediating; Membrane; Mentors; Microscopic; Microscopy; Microtubule-Organizing Center; Microtubules; Molecular; Morphogenesis; Mothers; Mutant Strains Mice; Mutation; Names; Organelles; Orofaciodigital Syndromes; Phase; Play; Process; Proteins; Research; Role; Sensory; Signal Transduction; Structure; Testing; Tissues; Training; Use of New Techniques; base; biophysical techniques; brain malformation; career; ciliopathy; cilium biogenesis; developmental disease; experimental study; genetically modified cells; human disease; innovation; nanometer resolution; nanoscale; novel; novel strategies; protein complex; reconstitution; recruit; trafficking

PROJECT SUMMARY/ ABSTRACT The heart of the centrosome, the microtubule organizing center, is composed of two centrioles. The two centrioles are not equal. The older of the two, called the mother centriole, differs structurally from the younger, daughter centriole. Mother centriole-specific structures confer the unique capability to nucleate the primary cilium, an organelle that serves as the cell’s antenna. Consequently, defects in centriolar proteins can cause human ciliopathies, diseases caused by disrupted ciliary function. Despite being universal features of vertebrate cells, how the mother and daughter centrioles differ and how centrioles are built remain mysterious. I uncovered a complex of proteins comprised of CEP90, MNR and OFD1 (which I have named DISCO for DIStal Centriole cOmplex) required for proper centriole morphogenesis. Mutations in DISCO components cause Joubert and Orofaciodigital syndromes, disorders of brain, face and limb development. By studying this novel centriolar complex, I seek to understand how centrioles are built, and how they are remodeled to support cilium assembly. Using an innovative combination of expansion and structured illumination microscopy (Ex-SIM), I will define how components of this complex structure the distal centriole and how human disease-associated mutations disrupt this sub-compartment (Aim 1). MNR and OFD1 control centriole length by an unknown mechanism. Using super-resolved imaging and biochemical assays, I will uncover molecular mechanisms by which centriole length is established by MNR and OFD1 (Aim 2). CEP90 and MNR are also components of centriolar satellites, poorly understood membrane-less granules surrounding the centrosome. I have found that centriolar satellites display hallmarks of phase separation. Using live-imaging and in vitro biochemical reconstitution, I will test the hypothesis that CEP90, MNR and OFD1 are trafficked to the centriole by phase- separated centriolar satellites to support ciliogenesis (Aim 3). With the help of an outstanding advisory committee, I will train in advanced imaging and biophysical techniques that will allow me to address fundamental questions on how centrioles and cilia are built. Spanning both the mentored and independent phases, these studies will illuminate how human disease-associated proteins build and modify centrioles to allow cilium biogenesis, and create a strong foundation for an independent research career studying the role of centrioles and cilia in human development and disease.

项目概要/摘要 中心体的心脏，即微管组织中心，由两个中心粒组成。 两个中心粒中较老的中心粒（称为母中心粒）在结构上与较年轻的中心粒不同。 子中心粒的特定结构赋予了初级中心粒成核的独特能力。 纤毛是一种作为细胞天线的细胞器，经过测试，中心粒蛋白的缺陷可能会导致这种现象。 人类纤毛病是由纤毛功能受损引起的疾病，尽管这是脊椎动物的普遍特征。 我发现，在细胞中，母体和子体中心粒的差异以及中心粒的构建方式仍然是个谜。 由 CEP90、MNR 和 OFD1 组成的蛋白质复合物（我将其命名为 DISCO，表示 DIStal Centriole DISCO 成分中适当的中心粒形态发生所需的突变导致 Joubert 和 通过研究这种新颖的中心粒，可以治疗口面部综合征、大脑、面部和肢体发育障碍。 复杂，我试图了解中心粒是如何构建的，以及它们如何被重塑以支持纤毛 使用扩展和结构照明显微镜 (Ex-SIM) 的创新组合，我将 定义这种复杂的成分如何构造远端中心粒以及人类疾病如何相关 突变破坏了这个子区室（目标 1），MNR 和 OFD1 通过未知的方式控制中心粒长度。 使用超分辨成像和生化检测，我将通过以下方式揭示分子机制。 中心粒长度由 MNR 和 OFD1 确定（目标 2）。 中心粒卫星，我发现围绕中心体的无膜颗粒知之甚少。 使用实时成像和体外生物化学，中心粒卫星显示出相分离的特征。 重构，我将测试 CEP90、MNR 和 OFD1 通过阶段运输到中心粒的假设 在杰出顾问的帮助下分离中心粒卫星以支持纤毛发生（目标 3）。 委员会，我将接受先进成像和生物物理技术的培训，这将使我能够解决基本问题 关于中心粒和纤毛如何构建的问题，涵盖指导阶段和独立阶段。 研究将阐明人类疾病相关蛋白如何构建和修改中心粒以允许纤毛 生物发生，并为研究中心粒作用的独立研究生涯奠定坚实的基础 和纤毛在人类发育和疾病中的作用。