Genetic Dissection of Signaling and Cilia

信号传导和纤毛的基因剖析

• 批准号: 10809398
• 负责人: TAMARA J. CASPARY
• 金额: $ 1.16万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-01-01 至 2027-12-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10809398
• 关键词: Affect; Base Pairing; Cilia; Cilium Microtubule; Comprehension; Disease; Dissection; Engineering; Family; Genetic; Genetic Screening; Guanosine Triphosphate Phosphohydrolases; Individual; Knowledge; Link; Molecular Genetics; Mus; Mutant Strains Mice; Mutation; Organelles; Paint; Pathway interactions; Process; Proteins; Regulation; Resolution; Role; SHH gene; Scientist; Series; Signal Pathway; Signal Transduction; Structure; Variant; cell type; cilium biogenesis; experimental study; hedgehog signal transduction; in vivo; interest; mouse model; mutant; neurodevelopment; smoothened signaling pathway; virtual

Project Summary/Abstract Cilia sparked phenomenal interest as scientists recognized them as a fundamental cellular organelle required for signaling. Untangling the specific mechanisms that regulate Sonic hedgehog (Shh) signaling within the cilium is difficult since so many mutants that disrupt ciliogenesis also affect Hh signaling. We have long focused on a small ciliary GTPase, ARL13B, that we hypothesize integrates the regulation of ciliogenesis and Hh signaling through distinct effectors and their downstream pathways. As a GTPase, single basepair mutations within the GTPase domain of ARL13B are predicted to disrupt individual effector pathways. ARL13B is highly enriched in cilia. By engineering mouse expressing only an ARL13B variant that does not localize to cilia, we genetically uncoupled the role of ARL13B in ciliogenesis from its role in signaling. We focus on mammalian neural development and through forward genetic screens identified mouse mutants in several proteins related to ARL13B. Additionally, other proteins in the ARL family of GTPases are implicated in cilia and signaling through what appear to be ARL13B related mechanisms. In the next five years, we propose using mouse mutants to define the regulatory relationships among these players in vivo and in specific cell types. These experiments will unravel ARL13B function in ciliogenesis, traffic of proteins to/within cilia, and Shh signal transduction at unprecedented resolution. Thus, our proposal will generate a molecular genetic toolkit from which the field will be poised to distinguish the regulation of cilia from that of Hh signaling. This is important to our fundamental understanding of cilia, ciliogenesis and cilia structure, as well as our basic comprehension of the Hh pathway.

项目概要/摘要 纤毛引起了科学家的极大兴趣，因为科学家们认识到它们是一种基本的细胞器 信令所需。解开调节 Sonic Hedgehog (Shh) 信号传导的具体机制 纤毛的研究很困难，因为许多破坏纤毛发生的突变体也会影响 Hh 信号传导。我们有很长的 专注于一种小的纤毛 GTP 酶，ARL13B，我们假设它整合了纤毛发生的调节和 Hh 信号传导通过不同的效应器及其下游途径。作为 GTP 酶，单碱基对 ARL13B GTPase 结构域内的突变预计会破坏个体效应通路。 ARL13B 纤毛高度丰富。通过工程改造仅表达 ARL13B 变体的小鼠，该变体不定位于 纤毛，我们从基因上将 ARL13B 在纤毛发生中的作用与其在信号传导中的作用分开。我们专注于 哺乳动物神经发育并通过正向遗传筛选鉴定了小鼠的几个突变体 与 ARL13B 相关的蛋白质。此外，GTPase ARL 家族中的其他蛋白质也与纤毛有关 并通过似乎与 ARL13B 相关的机制发出信号。未来五年，我们建议 使用小鼠突变体来定义这些参与者在体内和特定细胞中的调节关系 类型。这些实验将揭示 ARL13B 在纤毛发生、纤毛内/纤毛内的蛋白质运输以及纤毛发生中的功能。 嘘信号转导具有前所未有的分辨率。因此，我们的建议将产生分子遗传学 工具包，该领域将准备区分纤毛的调节和 Hh 信号传导的调节。这是 对于我们对纤毛、纤毛发生和纤毛结构的基本理解以及我们的基本知识非常重要 Hh 途径的理解。