Mechanisms underlying centriole morphogenesis

中心粒形态发生的机制

• 批准号: 10549843
• 负责人: Dhivya Kumar
• 金额: $ 3.22万
• 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 2023-03-24
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10549843
• 关键词: Ablation; 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; 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; restraint; superresolution imaging; 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.

项目总结/摘要 中心体的心脏，微管组织中心，由两个中心粒组成。两 中心粒并不相等。两者中较年长的称为母亲中心粒，在结构上与年轻的不同， 女儿中心粒母亲中心粒特异性结构赋予了独特的能力，使初级 纤毛，一种作为细胞天线的细胞器。因此，中心粒蛋白质的缺陷可以导致 人类睫状体病，由睫状体功能破坏引起的疾病。尽管是脊椎动物的普遍特征 然而，母中心粒和子中心粒的区别以及中心粒是如何形成的仍然是个谜。我发现了 由CEP 90、MNR和OFD 1组成的蛋白质复合物（我将其命名为DISCO，意思是DIStal Centriole cOmplex）是正确的中心粒形态发生所必需的。DISCO组分中的突变导致Joubert和 口面指综合征，大脑、面部和肢体发育障碍。通过研究这种新的中心粒 复杂，我试图了解中心粒是如何建立的，以及它们是如何改造，以支持纤毛 组装件.使用扩展和结构照明显微镜（Ex-SIM）的创新组合，我将 定义这个复杂结构的组成部分如何构成远端中心粒，以及人类疾病相关的 突变破坏了该亚区室（Aim 1）。MNR和OFD 1通过一种未知的方式控制中心粒长度 机制使用超分辨成像和生化分析，我将揭示分子机制， 其中中心粒长度由MNR和OFD 1确定（Aim 2）。CEP 90和MNR也是 中心粒卫星，知之甚少的无膜颗粒周围的中心体。我发现 中心粒卫星显示出相分离的特征。利用实时成像和体外生化 重建，我将测试的假设，CEP 90，MNR和OFD 1被贩运到中心粒的阶段- 分离中心粒卫星以支持纤毛发生（Aim 3）。在一位杰出顾问的帮助下 委员会，我将培训先进的成像和生物物理技术，使我能够解决基本的 中心粒和纤毛是如何形成的跨越辅导和独立阶段，这些 研究将阐明人类疾病相关蛋白如何构建和修饰中心粒， 生物起源，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。