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

基因

• 批准号: 3896312
• 负责人: R A KAHN
• 金额: --
• 依托单位: DIVISION OF CANCER TREATMENT
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/3896312
• 关键词: ADP ribosylation; Xenopus; adenylate cyclase; biological signal transduction; biotechnology; cell differentiation; cell growth regulation; gene expression; genetic library; genetic manipulation; guanine nucleotide binding protein; microinjections; molecular cloning; nucleic acid sequence; protein metabolism; protein structure function; yeasts

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.

ARF是一种新近鉴定的GTP结合调节蛋白 腺苷酸环化酶系统的组成部分。 这些研究旨在 在阐明ARF在信号转导中的作用时， 分化和增殖。 我们使用牛ARF 基因，从酵母中克隆两个同源基因ARFI和ARF 2， S. Cerevisiae. ARF 1的破坏导致生长缓慢，寒冷 敏感性和对氟化物的超敏感性。 我们计划 研究ARF 2破坏单独和 与ARF 1的破坏组合。 arf 1的抑制因子- 表型已经获得并将进行分析。 预计 其中一些抑制基因会编码其他蛋白质 在ARF通路中。 我们将确定人类ARF的能力， 拯救arf 1-表型以测试ARF功能的保守性。 此外，我们正在克隆和测序ARF从人类和 异光菌文库。 人类基因将被用来筛选 各种人类肿瘤中是否存在异常ARF 表情 非洲爪蟾基因将用于ARF的研究 在非洲爪蟾卵母细胞中的功能，其允许显微注射纯化的 ARF蛋白和由此产生的变化的生物化学测定， 细胞代谢