THE ARF FAMILY AS REGULATORS OF PROTEIN SECRETION AND OTHER CELLULAR PROCESSES

ARF 家族作为蛋白质分泌和其他细胞过程的调节剂

• 批准号: 3838040
• 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/3838040
• 关键词: ADP ribosylation; Golgi apparatus; carcinogenesis inhibitor; cell growth regulation; cellular pathology; cellular respiration; gene deletion mutation; genetic regulation; guanine nucleotide binding protein; guanosinetriphosphatases; human tissue; membrane fusion; membrane proteins; molecular cloning; nucleic acid sequence; oncoproteins; protein purification; protein structure function; protooncogene; secretion; suppressor mutations; yeasts

This section is involved in studies of the 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. The ARF proteins are a subfamily of structurally and functionally conserved 21 kDa GTP binding proteins. ARF has now been shown to have essential functions in the maintenance of the integrity and proper functioning of the Golgi complex. Current work has focused on characterizing the members of this family with regard to sequence, cellular location, and functions. We have discovered that in humans this family consists not only of the five previously identified gene products, but of more than 10 structurally related genes and their products. We are engaged in cloning and sequencing this newly enlarged family to begin to document its size, functions and importance to cell physiology and pathophysiology. In addition, as sequence information becomes available we continue to produce more specific antisera for diagnostic purposes and immunolocalization. We have continued to exploit simpler eukaryotic organisms, particularly yeast, to identify new functions for ARF proteins. We will perform functional screens to identify extragenic suppressors of ARF mutations in efforts to identify other components of the ARF pathway and potential regulators of ARF activities. ARF was found to be required for sporulation in this organism and ARF2 is much more potent than ARF1 in this activity. Expression of a number of mutants in yeast has also served as background for similar studies now underway to express wild-type and mutant ARF proteins in mammalian cells in culture. An exhaustive screen has now identified at least one temperature sensitive allele of ARF1 which will be used to assist in defining functions of ARF, consequences of an arf null cell, and functional screens for extragenic suppressors of the arf null cell.

本部分涉及功能和机制的研究 调节性 GTP 结合蛋白超家族成员的作用； 特别是 ADP-核糖基化因子 (ARF) 家族的成员。 ARF 蛋白是结构和功能上的一个亚家族 保守的 21 kDa GTP 结合蛋白。 ARF现已被证明具有 维护完整性和正确性的基本职能 高尔基复合体的功能。 目前的工作重点是 描述该家族成员的序列特征， 细胞的位置和功能。 我们发现，在人类中，这个家族不仅由 之前已鉴定出 5 种基因产物，但数量超过 10 种 结构相关的基因及其产物。 我们从事克隆 对这个新扩大的家族进行测序以开始记录其规模， 功能以及对细胞生理学和病理生理学的重要性。 在 此外，随着序列信息的出现，我们将继续 生产用于诊断目的的更特异的抗血清 免疫定位。 我们继续开发更简单的真核生物，特别是 酵母，以确定 ARF 蛋白的新功能。 我们将表演 功能筛选以鉴定 ARF 突变的基因外抑制因子 努力确定 ARF 途径的其他组成部分和潜力 ARF 活动的监管机构。 发现需要 ARF 在这种生物体中形成孢子，ARF2 比 ARF1 更有效 这项活动。 许多突变体在酵母中的表达也已 作为目前正在进行的类似研究的背景，以表达 培养的哺乳动物细胞中的野生型和突变型 ARF 蛋白。 一个 详尽的屏幕现已识别出至少一种温度敏感的 ARF1 的等位基因将用于协助定义 ARF 的功能， arf 无效细胞的后果以及外源性的功能筛选 arf 无效细胞的抑制子。