FUNCTIONAL CHARACTERIZATION OF ARF REGULATORY FACTORS

这个项目的主要目标是阐明细胞 腺苷二磷酸核糖化因子家族的功能 它们的调控机制。正如Brefeldin A(BFA)最近 被证明影响ARF功能和细胞分布，我们最近 扩大了我们的努力，包括更好地了解该机制 博鳌亚洲论坛的行动。 已经开发了ARF活性的特定抑制剂来提供 ARF蛋白在多种细胞内的作用的证据 膜事件。抗ARF多肽被发现是一种有效的抑制剂。 内质网高尔基体转运、高尔基体内转运和内体-内体 核聚变。此外，每种化验方法的灵敏度以及 核膜融合到GTP-GammaS被证明是由ARF介导的 蛋白质。 对ARF活动监管机构的搜索发现了一项活动 高尔基体膜似乎能激活ARF。此ARF正在激活 蛋白质(AAP)正被研究为一种潜在的作用部位 Brefeldin A及其作为蛋白质调节因子的潜在作用 分泌途径。Brefeldin A改变Brefeldin结构的能力 特异性外壳蛋白(ARF和Beta-COP)的高尔基体和定位是 正在与Brefeldin A作为一种 潜在的抗癌剂。 一种能够与ARF特异性结合并促进 ARF1的GTP酶活性也已被鉴定，并部分地 纯净的。这个ARF-GAP(GTP酶激活蛋白) 研究既是ARF的潜在效应因素，也是其下调因素 行为。

The main objectives of this project are the elucidation of the cellular functions of the family of ADP-ribosylation factors (ARFs) and the mechanism of their regulation. As Brefeldin A (BFA) has recently been shown to affect ARF functions and cellular distribution, we have recently expanded our efforts to include a better understanding of the mechanism of BFA action. Specific inhibitors of ARF activities have been exploited to provide evidence for the role of ARF proteins in a variety of intracellular membrane events. Anti-ARF peptides were found to be potent inhibitors of ER-Golgi transport, intra-Golgi transport, and endosome-endosome fusion. In addition, sensitivity of each of these assays, as well as nuclear membrane fusion, to GTPgammaS was shown to be mediated by ARF proteins. A search for regulators of ARF activity has identified an activity on Golgi membranes which appears to activate ARF. This ARF activating protein (AAP) is being investigated as a potential site of action of Brefeldin A and for its potential role as regulator of the protein secretory pathway. The ability of Brefeldin A to alter the structure of the Golgi and location of specific coat proteins (ARF and Beta-COP) is being investigated in concert with the development of Brefeldin A as a potential anti-cancer agent. A protein with the ability to specifically bind ARF and promote the GTPase activity by ARF1 has also been identified and is partially purified. This ARF-GAP (GTPase activating protein) is under investigation as both potential effector and down-regulator of ARF actions.