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.

项目摘要/摘要 当科学家认为它们是基本的蜂窝细胞器时，纤毛引发了惊人的兴趣 信号所需的。解开调节声音刺猬（SHH）信号的特定机制 纤毛很困难，因为破坏纤毛发生的许多突变体也会影响HH信号传导。我们有很长的时间 专注于小型睫状GTPase ARL13B，我们假设我们整合了纤毛发生和 HH通过不同的效应子及其下游途径发出信号。作为GTPase，单基台 ARL13B的GTPase结构域内的突变预计会破坏单个效应器途径。 arl13b 在纤毛中高度丰富。通过工程鼠标仅表达不本地化的ARL13B变体 纤毛，我们从基因上脱离了Arl13b在纤毛发生中的作用，从其在信号传导中的作用中。我们专注于 哺乳动物神经发育并通过正向遗传筛选确定了几种小鼠突变体 与ARL13B有关的蛋白质。此外，GTPases ARL家族中的其他蛋白质与纤毛有关 并通过似乎与ARL13B相关的机制发出信号。在接下来的五年中，我们建议 使用小鼠突变体来定义这些玩家在体内和特定细胞中的调节关系 类型。这些实验将在纤毛生成，蛋白质到/纤毛内的流量以及 史无前例的分辨率下的SHH信号转导。因此，我们的建议将产生分子遗传 该田地将从该工具包中区分纤毛的调节与HH信号的调节。这是 对于我们对纤毛，纤毛生成和纤毛结构的基本理解以及我们的基本 HH通路的理解。