GENETIC & IMMUNOLOGIC ANALYSES OF THE ADP-RIBOSYLATION FACTOR, ARF

ARF是最近发现的一种GTP结合调节蛋白 腺苷环化酶系统的组成部分。这些研究的目的是 在阐明ARF在信号转导中的作用时， 分化和增殖。我们已经使用了牛ARF 从酵母中克隆两个同源基因ARFI和ARF2， 酿酒酵母。ARF1基因中断导致生长缓慢，寒冷 对氟化物的敏感性和超敏性。我们计划 调查ARF2中断单独和在 与ARF1的干扰相结合。ARF1的抑制子- 已获得表型，并将进行分析。这是意料之中的 其中一些抑制基因将编码其他蛋白质 在ARF途径中。我们将确定人类ARF的能力 挽救arf1表型以检测ARF功能的保守性。 此外，我们正在克隆和测序人类和人类的ARF Xenoous laevis图书馆。人类基因将被用于筛查 多种人类肿瘤中存在的异常ARF 表情。非洲爪哇基因将用于ARF的研究 非洲爪哇卵母细胞的功能允许微量注射纯化的 ARF蛋白及其所致变化的生化分析 细胞新陈代谢。

ARF is a recently characterized GTP-binding regulatory protein component of the adenylate cyclase system. These studies are aimed at the elucidation of the role of ARF in signal transduction, differentiation, and proliferation. We have used the bovine ARF gene to clone two homologous genes, ARFI and ARF2, from the yeast, S. Cerevisiae. Disruption of ARF1 leads to slow growth, cold sensitivity, and supersensitivity to fluoride. We plan to investigate the consequences of ARF2 disruption both alone and in combination with disruption of ARF1. Suppressors of the arf1- phenotype have been obtained and will be analyzed. It is expected that some of these suppressor genes will code for other proteins in the ARF pathway. We will determine the ability of human ARF to rescue the arf1- phenotype to test for conservation f ARF function. In addition, we are cloning and sequencing ARF from both human and Xenoous laevis libraries. The human gene will be used to screen a variety of human tumors for the presence of abnormal ARF expression. The Xenopus gene will be used in studies of ARF function in Xenopus oocytes which allow microinjection of purified ARF proteins and biochemical assays of resultant changes in cellular metabolism.