CHARACTERIZATION OF THE ARF FAMILY OF REGULATORY GTP-BINDING PROTEINS

这个项目涉及到对结构、功能和机制的研究。 调控GTP结合超家族成员的作用 蛋白质，特别是ADP核糖化因子(ARF)的成员 一家人。ARF已被证明在 维持内质的完整性和正常运作 网状结构、高尔基复合体和内吞体室。ARF蛋白也 刺激磷脂酶D，很可能是直接激活 这个效应器。 目前有6种哺乳动物ARF蛋白和6种哺乳动物ARF样蛋白 蛋白质完全克隆和测序。新奇基因的克隆和测序 这个家族的成员已经放慢了速度，以便有更多的时间来分析功能 已知成员的名单。我们利用了许多功能屏幕 为ARF蛋白在酵母中的作用提供新的线索 真核生物。这些筛查包括抑制者分析，一个 ARF1的条件致死筛选和双杂交筛选鉴定 ARF信号通路中的其他成分。这些方法中的每一种 导致了人类和酵母基因的克隆，这些基因的产物相互作用 与Arf蛋白结合。这些分析的结果可能会打开新的 对这一日益重要的细胞家族的研究领域 监管者。 解决了ArF国内生产总值的三维晶体结构。工作 继续研究活性的ARF GTP和肉豆蔻酰化的ARF结构 应有助于确定关键功能域和设计 不同Arf活性的特定抑制剂。

This project involves studies of the structures, functions and mechanisms of action of members of the superfamily of regulatory GTP binding proteins; particularly the members of the ADP-ribosylation factor (ARF) family. ARF has been shown to have essential functions in the maintenance of the integrity and proper functioning of the endoplasmic reticulum, Golgi complex, and endosome compartments. ARF proteins also stimulate phospholipase D, likely serving as the direct activators of this effector. There are currently 6 mammalian ARF proteins and 6 mammalian ARF-like proteins fully cloned and sequenced. The cloning and sequencing of novel members of this family has slowed to allow more time to analyze functions of the known members. We have utilized a number of functional screens in yeast to provide new clues to the actions of ARF proteins in eukaryotes. These screens have included suppressor analysis, a conditional lethal screen of ARF1, and two-hybrid screening to identify other components in ARF signaling pathways. Each of these approaches have led to the cloning of human and yeast genes whose products interact with Arf proteins. Results of these analysis are likely to open up new areas of research into this increasingly important family of cellular regulators. The three dimensional crystal structure of ARF GDP has been solved. Work continues on the active, ARF GTP and myristoylated ARF structures as each should aid in the determination of critical functional domains and design of specific inhibitors of different Arf activities.