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.

项目摘要/摘要 中心体的心脏是微管组织中心，由两个中心元素组成。 中心数不相等。两者中的年龄较大，称为母亲中心，在结构上与年轻人区分开 女儿Centriole。母亲中心特异性结构会议会议核能核能的独特能力 cilium，是一个细胞天线的细胞器。因此，中心蛋白的缺陷会导致 人纤毛病，由睫状功能中断引起的疾病。尽管是脊椎动物的通用特征 细胞，母女的中心含量如何不同以及如何建立中心元仍然神秘。我发现了 CEP90，MNR和OFD1积累的蛋白质复合物（我将其命名为Disco distal Centriole 复杂）适当的中心形态发生所需。迪斯科组件中的突变导致乔伯特和 Orofaciodigital综合征，大脑，面部和肢体发育的疾病。通过研究这个新颖的中心 复杂的是，我试图了解中心元素的建造方式，以及如何改建以支持纤毛 集会。使用扩展和结构化照明显微镜（EX-SIM）的创新组合，我将 定义这种复杂结构的远端中心的组成部分以及与人类疾病相关的如何 突变破坏了该子室（AIM 1）。 MNR和OFD1控制中心长度未知 机制。使用超级分辨的成像和生化测定，我将通过 MNR和OFD1建立了哪个中心长度（AIM 2）。 CEP90和MNR也是 中心卫星，不了解中心体周围的无膜颗粒。我发现 中心卫星显示相位的标志。使用现场模仿和体外生化 重组，我将检验以下假设：CEP90，MNR和OFD1通过相位被贩运到Centriole 分离的中心卫星以支持纤毛发生（AIM 3）。借助出色的咨询 委员会，我将培训高级成像和生物物理技术，这将使我能够解决基本 有关中心和纤毛的建造方式的问题。跨越修补阶段和独立阶段，这些 研究将阐明与人类疾病相关的蛋白质如何构建和修饰中心元素以允许纤毛 生物发生，并为研究中心的作用的独立研究职业创造了坚实的基础 和纤毛在人类发展和疾病中。