Molecular mechanisms of ARF family GTPases

项目概要/摘要 调节性GTP酶的ARF家族成员在细胞信号传导中起节点的作用， 协调基本细胞过程;包括膜运输，能量代谢， 纤毛发生和细胞骨架。我研究了第一个ARF和后来的ARF样（ARL）蛋白， 30多年来，利用生物化学、细胞和分子生物学、遗传学和 系统发育的方法，并建议继续这些研究，重点是他们的行动 与纤毛和中心体相关，以便对五个子集进行更详细的机制研究 这个家族的30个GTP酶我们将使用基因组编辑来产生一个敲除的集合， 细胞系来破译所研究的蛋白质的信号。我们将使用经典的蛋白质 生物化学来纯化和发现这些途径中的新组分。我们都要学习 这五种GTP酶中的每一种在细胞中的功能，并与它们的体外生物化学研究相关联。 酶学我们将探索细胞信号传导的串扰或更高层次有序化的可能性， 并为非典型GTP酶开发新的单一作用，作为确定共享 或是家庭中的独特机制更好地了解这些系统将揭示 对细胞生物学基本方面的新见解，以及为 干预以改变人类疾病的进程;包括但不限于癌症，心脏病， 疾病、神经变性、睫状体病、视网膜变性和耳聋。

Project Summary/Abstract Members of the ARF family of regulatory GTPases function as nodes in cell signaling to coordinate essential cell processes; including membrane traffic, energy metabolism, ciliogenesis, and the cytoskeleton. I have studied first ARF and later ARF-like (ARL) proteins for over 30 years, using a combination of biochemical, cell and molecular biological, genetic, and phylogenetic approaches and propose to continue these studies with a focus on their actions linked to cilia and centrosomes to allow more detailed mechanistic studies of a subset of five of the 30 GTPases in this family. We will use genome editing to generate a collection of knockout cell lines to decipher signaling by the proteins under study. And we will use classical protein biochemistry to purify and discover novel components in these pathways. We will both study functions of each of these five GTPases in cells, and link to in vitro biochemical studies of their enzymology. We will explore the potential for cross-talk or higher level ordering of cell signaling, and also develop novel, single actions for atypical GTPases as a means of determining shared or unique mechanisms within the family. A better understanding of these systems will reveal novel insights into fundamental aspects of cell biology as well as providing potential targets for intervention to alter the course of human diseases; including but not limited to cancer, heart disease, neurodegeneration, ciliopathies, retinal degeneration, and deafness.