Functoinal compartmentalization of hedgehog signal transduction in primary cilia

初级纤毛中刺猬信号转导的功能划分

• 批准号: 9388856
• 负责人: Tim Stearns
• 金额: $ 30.81万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-15 至 2021-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9388856
• 关键词: Adult; Behavior; Biochemical; Biochemical Genetics; Cells; Cellular biology; Centrosome; Cholesterol; Cilia; Competence; Complex; Congenital Abnormality; Defect; Development; Diffusion; Embryonic Development; Erinaceidae; Event; Excision; Genetic Epistasis; Goals; Image; In Vitro; Inherited; Label; Ligands; Link; Malignant Neoplasms; Medical; Membrane; Methods; Microscopy; Molecular; Molecular Genetics; Motion; Movement; Natural regeneration; Optics; Organ; Organelles; Pathway interactions; Phenotype; Physiology; Process; Proteins; Resolution; Role; Signal Transduction; Signaling Protein; Structure; Testing; Tissues; Transducers; Work; base; cell type; cellular imaging; ciliopathy; experimental study; extracellular; functional gain; functional loss; gain of function; genetic approach; hedgehog signal transduction; high resolution imaging; human disease; imaging genetics; in vivo; insight; kinetosome; lysosomal proteins; novel; particle; protein complex; protein function; residence; segregation; single molecule; smoothened signaling pathway; tissue regeneration; trafficking

Functional Compartmentalization of Hedgehog Signal Transduction in Primary Cilia Abstract Primary cilia are small, antenna-like organelles critical for vertebrate development and physiology. Defects in cilia result in human diseases called ciliopathies, characterized by a wide spectrum of phenotypes, highlighting their important role in multiple cell types and organs. Although not completely surrounded by a membrane, cilia form a distinct compartment that receives and transmits extracellular signals. Their function critically depends on the dynamic changes in protein composition and localization. The Hedgehog (Hh) signaling transduction, essential for embryonic development, adult tissue regeneration and cancer, largely takes place at primary cilia. The Hh-transduction proteins Smoothened (Smo), Patched1 (Ptch1), Suppressor of Fused (SuFu) and Gli2 shuttle into and out of cilia when cells receive the Hh signal, and changes in localization correlate with pathway activation. The molecular basis for most of these steps is not clear. In this proposal, we will build upon our previous work in Hh signal transduction and in the cell biology of cilia and centrosomes, to answer unresolved questions about protein compartmentalization in cilia and Hh signal transduction. We will apply high-resolution optical microscopy, molecular genetics and novel biochemical methods to determine how specific interactions with ciliary protein complexes control Hh-transducer dynamics in cilia, and, more broadly, how ciliary responsiveness is established; investigate how Ptch1 interactions within the cilium regulate Smo activation; finally, we will elucidate how removal of Hh-transducers from cilia is controlled, extending our recent discovery of Rilp-like proteins as regulators of this process. In summary, we propose to establish a high-resolution, spatially and temporally resolved picture of Hedgehog signaling in the primary cilium. We believe that only by having such an in- depth analysis will it be possible to understand the molecular basis of Hedgehog signaling. !

Hedgehog信号转导的功能区室化 初级纤毛 摘要 初级纤毛是小的触角状细胞器，对脊椎动物的发育至关重要， physiology.纤毛缺陷会导致称为纤毛病的人类疾病，其特征是广泛的 表型谱，突出了它们在多种细胞类型和器官中的重要作用。 虽然纤毛没有完全被膜包围，但纤毛形成了一个独特的隔室， 接收和传输细胞外信号。它们的功能主要取决于 蛋白质组成和定位的变化。Hedgehog（Hh）信号转导， 对于胚胎发育、成年组织再生和癌症至关重要， 初级纤毛Hh-转导蛋白Smoothened（Smo），Patched 1（Ptch 1）， 当细胞接受Hh时，SuFu和Gli 2穿梭进出纤毛 信号和定位变化与途径活化相关。的分子基础 这些步骤大多不清楚。在本提案中，我们将以Hh中以前的工作为基础， 信号转导和纤毛和中心体的细胞生物学，以回答悬而未决的 纤毛中蛋白质区室化和Hh信号转导的问题。我们将应用 高分辨率光学显微镜，分子遗传学和新的生化方法， 确定与纤毛蛋白复合物的特异性相互作用如何控制Hh-转换器 纤毛动力学，更广泛地说，纤毛反应是如何建立的;研究如何 纤毛内的Ptch 1相互作用调节Smo激活;最后，我们将阐明如何 从纤毛中去除Hh-转换器是受控的，扩展了我们最近发现的类HLP 蛋白质作为这一过程的调节剂。 总之，我们建议建立一个高分辨率，空间和时间分辨 初级纤毛中Hedgehog信号的图片。我们相信，只有拥有这样一个- 深入分析将有可能理解Hedgehog信号传导的分子基础。 !