Generation of Diverse Centrosomes, Cilia and Flagellae During Development

发育过程中多种中心体、纤毛和鞭毛的产生

• 批准号: 10590581
• 负责人: David M Glover
• 金额: $ 50.21万
• 依托单位: CALIFORNIA INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-05-15 至 2025-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10590581
• 关键词: Affect; Applied Genetics; Biology; Brain; Cell Cycle; Cell Line; Cell division; Cells; Centrioles; Centrosome; Cerebral cortex; Cilia; Cilium Microtubule; Complex; Cultured Cells; Cytoplasm; Defect; Development; Disease; Drosophila genus; Eukaryota; Flagella; Foundations; Functional disorder; Future; G2 Phase; Generations; Genetic; Germ Lines; Growth; Heritability; Human; Inherited; Interphase; Lead; Leg; Length; Light; Malignant Neoplasms; Meiosis; Microcephaly; Microtubule Triplet; Microtubules; Mitosis; Molecular; Molecular Biology; Motion; Mutagenesis; Mutation; Nomenclature; Organ; Organelles; Pathology; Pathway interactions; Patients; Phosphorylation; Physiology; Pilot Projects; Procentriole; Process; Prognosis; Proteins; Research; Role; Set protein; Signal Transduction; Somatic Cell; Specific qualifier value; Spermatocytes; Spermatogenesis; Structure; Testing; Tissues; Work; brain abnormalities; cell type; ciliopathy; cilium biogenesis; genetic approach; human disease; imaginal disc; kinetosome; male; man; neurosensory; organizational structure; protein function; recruit; sperm cell; stem cells; structural biology

PROJECT SUMMARY Centrioles are required to template cilia and flagella and to organize centrosomes that nucleate cytoplasmic microtubules in interphase and dividing cells. Centrosomes lose their ability to organise cytoplasmic microtubules when they become basal bodies, which can template the formation of primary cilia for signalling or flagellae for propelling sperm. To understand these diverse roles, it necessary to study centrioles in the development of different cell types. Yet, their replication, structure and function has been largely studied in cultured cells. Here we take advantage of the genetic and developmental advantages of Drosophila to study centriole function in two cell types; cells destined to make neurosensory cilia in the leg imaginal disc and male germ line cells destined to generate sperm. Our focus is on the roles of a network of centriole proteins in which Ana3, counterpart of human ROTATIN and Rcd4, PPP1R35 in human cells, interact with each other and with a set of proteins required for various steps of the elongation and maturation of the centriole necessary for it to become a basal body. Specifically, we will: (i) apply molecular and structural biology approaches to determine how the Ana3 and Rcd4 proteins interact with each other and with other centriole proteins to specify different aspects of centriole assembly and function and how this is regulated by phosphorylation. We will test the functions of these interactions through directed mutagenesis. (ii) use genetic approaches to determine inter-dependencies of the networks that regulate centriole elongation in the conventional cell division cycles of cultured cells and the leg imaginal disc and in cells that develop neurosensory cilia. (iii) determine the differential roles of Rcd4 and Ana3 and their interacting network of proteins in the multiple steps leading to formation of the giant centrioles of spermatocytes and their sequential roles in generating primary cilia and subsequently sperm flagellae., Because centriole duplication pathways are highly conserved between Drosophila and man, our findings will shed light onto a wide range of human diseases including cancer, where centrosome defects indicate poor prognosis; heritable ciliopathies, which affect centrioles and cilia in a wide range of tissues; and in microcephaly, where defects in proteins such as ROTATIN, whose counterpart we study here, lead to abnormal brain development.

项目摘要 中心粒是纤毛和鞭毛的模板，并组织成核的中心体 间期和分裂细胞中的细胞质微管。中心体失去了组织能力 细胞质微管，当他们成为基体，这可以模板的形成， 初级纤毛发出信号或鞭毛推进精子。为了理解这些不同的角色， 研究中心粒在不同细胞类型发育中的作用。然而，它们的复制，结构 在培养的细胞中大量研究了其功能。在这里，我们利用遗传和 果蝇的发育优势，研究中心粒功能的两种细胞类型;细胞注定要 使腿上的神经感觉纤毛成为成虫盘，使雄性生殖细胞注定要产生精子。 我们的重点是一个中心粒蛋白质网络的作用，其中Ana 3，对应的人类 ROTATIN和Rcd 4，PPP 1 R35在人类细胞中相互作用，并与一组蛋白质相互作用 所需的各种步骤的延伸和成熟的中心粒所必需的，它成为 基体具体而言，我们将：（i）应用分子和结构生物学方法来确定 Ana 3和Rcd 4蛋白如何相互作用以及与其他中心粒蛋白相互作用， 中心粒组装和功能的不同方面以及磷酸化如何调节中心粒组装和功能。我们 将通过定向诱变来测试这些相互作用的功能。(ii)使用遗传学方法 为了确定在传统的细胞内调节中心粒伸长的网络的相互依赖性， 培养细胞的细胞分裂周期和腿成虫盘以及发育神经感觉的细胞 纤毛。(iii)确定Rcd 4和Ana 3及其相互作用的蛋白质网络在 精母细胞巨大中心粒形成的多个步骤及其顺序 在产生初级纤毛和随后的精子鞭毛中的作用， 由于中心粒复制途径在果蝇和人类之间高度保守， 将为包括癌症在内的一系列人类疾病提供线索，其中中心体缺陷表明 预后不良;遗传性纤毛病，影响广泛组织中的中心粒和纤毛;以及 小头畸形，其中蛋白质缺陷，如旋转蛋白，我们在这里研究的对应物，导致 大脑发育异常