Biology of the ARL13B GTPase

ARL13B GTP 酶的生物学

基本信息

批准号：
8932018
负责人：
TAMARA J. CASPARY
金额：
$ 42.75万
依托单位：
EMORY UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-09-30 至 2018-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8932018
关键词：
Affect Alleles Animal Model Animals Anions Bardet-Biedl Syndrome Binding Binding Sites Biochemical Biochemical Genetics Biochemistry Biology Brain Cell Culture Techniques Cell Line Cells Cilia Cilium Microtubule Complex Data Defect Development Diffusion Disease Disease model Embryo Erinaceidae Eukaryotic Cell Family Fibroblasts Functional disorder G-Protein-Coupled Receptors GTPase-Activating Proteins Genes Genetic Genotype Guanine Nucleotides Guanosine Triphosphate Phosphohydrolases Health Hereditary Disease Human Human Genetics Hydrolysis Joubert syndrome Kidney Knockout Mice Lead Location Maintenance Mammalian Cell Missense Mutation Modeling Molecular Models Monomeric GTP-Binding Proteins Movement Mus Mutate Mutation Nucleotides Organism Output Pathway interactions Patients Pattern Phenotype Phosphoric Monoester Hydrolases Play Process Property Proteins Research Resolution Role Signal Transduction Solid Source Specificity Structure Study models Substrate Specificity System Testing Therapeutic Intervention Tissues VDAC1 gene Variant Vertebrates Work base cell type ciliopathy cilium biogenesis human disease in vivo in vivo Model interest member molecular modeling mouse development neuronal cell body novel protein purification protein transport response voltage

项目摘要

DESCRIPTION (provided by applicant): Cilia have sparked keen interest because of their proven importance in many human genetic diseases (collectively termed ciliopathies and including Joubert syndrome (JS)) and in vertebrate/mouse development; appreciation of their central role in aspects of cell signaling is growing, as well. Primary, immotile cilia are microtubule-based projections found on virtually every eukaryotic cell. We focus on the small GTPase, ARL13B, because its absence leads to defects in (1) cilia structure, (2) movement and localization of specific proteins to and (3) within cilia, and (4) patients with mutations in ARL13 have Joubert syndrome (JS), which involves renal, ocular, and brain anomalies. This application has four specific aims, in which we propose to (1) define the biochemical properties of ARL13B as a GTPase and identify and characterize novel effectors, (2) identify, purify, and characterize ARL13B GTPase activating proteins (GAPs), (3) determine the level of complexity in downstream ARL13B signaling and identify one or more effectors, (4) determine the tissue-specific consequences of JS causing ARL13B mutations in mice. We will use the complementary expertise of the two PIs to develop models for ARL13B signaling at cilia, with direct relevance to disease processes. We will pair a novel genetic deletion system in mouse embryo fibroblasts with detailed biochemical studies and modeling of ARL13B as a regulatory GTPase to develop molecular models for ARL13B's actions in cells and animals. Together, these approaches will allow us to identify and rigorously test downstream effectors and GTPase activating proteins (GAPs) of ARL13B actions in the development of the pathways affected by JS. Finally, by defining the in vivo, tissue specific function of the JS causative ARL13B mutations, we will both model the disease and identify the mechanism underlying tissue-specific Arl13b function and patient phenotypic variability. Together these aims will provide a solid experimental basis for establishing models of Arl13b actions, with clear implications for many aspects of cilia function, cell signaling, mammalian development, and human disease.

描述（由申请人提供）：Cilia激发了敏锐的兴趣，因为它们在许多人类遗传疾病（共同称为纤毛病，包括Joubert综合征（JS））和脊椎动物/小鼠发育中具有证实的重要性；对它们在细胞信号转导方面的核心作用也正在增长。原发性纤毛是基于微管的投影，几乎是每个真核细胞。我们专注于小的GTPase ARL13B，因为它的缺失会导致（1）纤毛结构中的缺陷，（2）特异性蛋白质在纤毛内的运动和定位，以及（4）ARL13突变的患者joubert综合征（JS），涉及肾脏，肾脏，眼睛，脑，眼睛，脑，大脑和大脑。该应用具有四个具体目标，在其中我们建议（1）将ARL13B的生化特性定义为GTPase，并识别和识别和表征新颖的效应子，（2）识别，纯化和表征ARL13B GTPase激活蛋白（GAP）（GAP）（GAP）（GAP）（GAP），（3）确定一个S-Sprem and snstream Arl13B Signallie andl13B的级别（4 JS引起小鼠ARL13B突变的of。我们将使用两个PI的互补专业知识来开发纤毛的ARL13B信号传导模型，并与疾病过程直接相关。我们将在小鼠胚胎成纤维细胞中的新遗传缺失系统与详细的生化研究和ARL13B作为调节性GTPase的建模，以开发ARL13B在细胞和动物中的作用的分子模型。这些方法将使我们能够识别并严格测试ARL13B作用的下游效应子和GTPase激活蛋白（GAP）在受JS影响的途径中的开发中。最后，通过定义JS病因ARL13B突变的体内组织特异性功能，我们将既对疾病进行建模并确定组织特异性ARL13B功能和患者表型变异性的基本机制。这些目标共同为建立ARL13B动作模型提供了可靠的实验基础，对纤毛功能，细胞信号传导，哺乳动物发育和人类疾病的许多方面具有明显的影响。