CHARACTERIZATION OF THE ARF FAMILY OF REGULATORY GTP-BINDING PROTEINS

GTP 调节蛋白 ARF 家族的表征

• 批准号: 3752321
• 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/3752321
• 关键词: ADP ribosylation; Golgi apparatus; Saccharomyces cerevisiae; alleles; biological signal transduction; cellular respiration; endoplasmic reticulum; eukaryote; fatty acylation; fungal genetics; genetic regulation; guanine nucleotide binding protein; human genetic material tag; human tissue; lethal genes; molecular cloning; myristates; northern blottings; nucleic acid sequence; phospholipase D; protein structure function; sporogenesis; transcription factor

This section is involved in studies of the structure, function and mechanism 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. The cloning and sequencing of novel members of the ARF family in humans and the yeast, S. cerevisiae, continues. There are currently 6 mammalian ARF proteins and 6 mammalian ARF-like proteins fully cloned and sequenced. In addition, we have partial sequence of at least five more members in yeast, and several more in humans. Yeast genetics continues to provide new clues to the actions of ARF proteins in eukaryotes. A conditional lethal allele of ARF1 revealed a role for ARF in sporulation. Analysis of another allele of arf1 led to the observation that ARF function is also required for respiration. It is likely that the roles for ARF proteins in these essential cellular processes provide important clues toward understanding the mechanisms of action of these critical regulatory proteins. The two-hybrid technique was used to identify other components in an ARF signaling pathway. This approach led to the cloning of 6 human genes whose products interact with human ARF3. Northern blots revealed that five of these 6 genes are expressed in a wide variety of human tissues, and one with a more limited tissue distribution. 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.

本部分主要研究了植物的结构、功能和 调节性GTP超家族成员的作用机制 结合蛋白;特别是ADP-核糖基化因子的成员 (ARF)家人 ARF已被证明具有重要的功能， 维持内质网的完整性和正常功能 内质网、高尔基复合体和内体隔室。 ARF蛋白还 刺激磷脂酶D，可能作为直接激活剂， 效应器ARF家族新成员的克隆和测序 人和酵母菌，S.酿酒，继续。 目前有6 哺乳动物ARF蛋白和6种哺乳动物ARF样蛋白完全克隆， 测序 另外，我们还有至少五个以上的部分序列 酵母中的成员，以及人类中的几个成员。酵母遗传学继续 为研究ARF蛋白在真核生物中的作用提供了新的线索。 一 ARF 1的条件致死等位基因揭示了ARF在孢子形成中的作用。 对arf 1的另一个等位基因的分析表明， 呼吸功能也需要。 很可能这些角色 对于ARF蛋白在这些基本细胞过程中提供重要的 了解这些关键因素的作用机制的线索 调节蛋白 双杂交技术用于鉴定其他 ARF信号通路中的一种成分。 这种方法导致了克隆 6个人类基因，其产物与人类ARF 3相互作用。 北方印迹法 揭示了这6个基因中的5个在各种各样的 一种是人体组织，另一种是组织分布更有限的组织。三 求解了ARF~GDP的三维晶体结构。 工作仍在继续 活性，ARF~GTP和豆蔻酰化ARF结构，因为每一个都应该有助于 在确定关键功能域和设计特定的 抑制剂的