MOLECULAR BASIS OF RAN MEDIATED NUCLEAR TRANSPORT

RAN 介导的核输运的分子基础

• 批准号: 2736959
• 负责人: ANITA H. CORBETT
• 金额: $ 21.65万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-02-01 至 2003-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2736959
• 关键词: G protein; Saccharomyces cerevisiae; biological signal transduction; fungal proteins; guanine nucleotide binding protein; guanosine diphosphate; guanosinetriphosphatases; intracellular transport; mutant; nuclear membrane; pore forming protein; protein protein interaction; protein structure function; protein transport; site directed mutagenesis; yeast two hybrid system

The broad long term objective of this proposal is to understand how the Ran GTPase drives bi-directional movement of macromolecules across the nuclear envelope. There is evidence to suggest that Ran cooperates with a number of accessory proteins to move proteins into and out of the nucleus. The specific aims of this proposal are: 1) to examine the in vivo interactions that are essential for Ran function; 2) to test the hypothesis that the essential cellular role of the Ran-GDP binding protein, nuclear transport factor 2 (NTF2) is to concentrate Ran-GDP at the nuclear pore, where Ran is required to initiate nuclear import; and 3) to use a genetic approach to define the site(s) on the nuclear pore that represent(s) the docking site for NTF2 and most probably the NTF2- Ran complex. The proposed studies use the budding yeast Saccharomyces cerevisiae as a model for in vivo genetic and cell biological experiments and extend to biochemical studies, in vivo functional studies, and cell biological experiments and extend to biochemical studies, in vivo functional studies, and structural studies of both the yeast proteins and their highly conserved human counterparts. The health-relatedness of this proposal is two-fold. First, activated signal transduction pathways send a signal to the nucleus in order to respond to stimuli and activate transcription. This is most often accomplished by the movement of a protein into the nucleus. This aspect of signaling is often ignored or trivialized, yet it may represent an unexploited targeted for blocking specific cellular signals as well as the unregulated signals that arise in transformed cells. Second, viruses that infect human cells exploit the endogenous nuclear transport machinery both to gain entry to the nucleus and later to rapidly export their own replicated genetic material. A more detailed understanding of the machinery that mediates nuclear transport may provide novel targets for anti-viral therapies.

该提案的广泛长期目标是了解如何 Ran GTPase 驱动大分子在整个细胞内的双向运动 核膜。有证据表明Ran与 许多辅助蛋白将蛋白质移入和移出 核。该提案的具体目标是： 1) 审查 Ran 功能所必需的体内相互作用； 2）测试 假设 Ran-GDP 结合的重要细胞作用 蛋白质，核转运因子2（NTF2）是将Ran-GDP浓缩在 核孔，需要 Ran 来启动核输入；和 3) 使用遗传方法来定义核孔上的位点 代表 NTF2 的对接位点，最有可能是 NTF2- 然复杂。拟议的研究使用芽殖酵母 Saccharomyces 酿酒酵母作为体内遗传和细胞生物学的模型 实验并扩展到生化研究、体内功能 研究和细胞生物学实验并扩展到生化 研究、体内功能研究和结构研究 酵母蛋白及其高度保守的人类对应物。这 该提案与健康的相关性有两个方面。一、激活信号 转导途径向细胞核发送信号以做出反应 刺激并激活转录。这通常是完成的 通过蛋白质进入细胞核的运动。信令的这方面 经常被忽视或轻视，但它可能代表着一种未开发的 旨在阻断特定的细胞信号以及 转化细胞中出现不受调节的信号。二、病毒 利用内源性核运输感染人类细胞 机械装置既可以进入核心，又可以快速输出 他们自己复制的遗传物质。更详细的了解 介导核运输的机制可能提供新的目标 用于抗病毒治疗。