NMR STUDIES OF THE MECHANISMS OF ACTION OF TRP-REPRESSOR

TRP 阻遏物作用机制的 NMR 研究

• 批准号: 2177013
• 负责人: OLEG JARDETZKY
• 金额: $ 22.05万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1985
• 资助国家: 美国
• 起止时间: 1985-07-01 至 1997-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2177013
• 关键词: Escherichia coli; bacterial DNA; chemical binding; genetic operator element; genetic regulation; microorganism genetics; molecular dynamics; molecular genetics; mutant; nuclear magnetic resonance spectroscopy; nucleic acid structure; point mutation; transcription factor; tryptophan

The objective of the proposed project is to achieve a detailed and precise understanding of the molecular mechanism of genetic regulation, at a level at which specific changes in the operation of this mechanism could be predicted and engineered. The ability to predict and control modifications to the function of genetic regulatory mechanisms will have far reaching consequences to the ability to treat genetic diseases and to use the mechanisms as targets for rational drug design, but existing knowledge is as yet insufficient to reach these goals. The trp-repressor from E. coli was chosen as a model system for detailed study, because it is one of the smallest and simplest complete regulatory systems available today. Starting with the known structures of the repressor protein, with and without the activator L-tryptophan, and of the ternary DNA-repressor-L-trp complex, it is possible to systematically investigate the role of each structural element (amino acid residue) in (l) maintaining specific contacts with the L-trp ligand and (2) with the operator DNA and (3) transmitting the signal regulating DNA binding between the binding sites. This will be achieved by studying point and multiple mutations which are known to alter the conformation and dynamics of the system. Very promising initial results have already been obtained with some of the mutants, providing clear directions for future studies. Correlation of the findings should allow an understanding of the signaling code and the design of the regulatory apparatus.

拟议项目的目的是实现详细而精确的 了解遗传调节的分子机制，在一个水平上 该机制的操作的具体变化可能是 预测和设计。预测和控制修改的能力 遗传调节机制的功能将达到遥远 对治疗遗传疾病和使用的能力的后果 机制是理性药物设计的目标，但现有知识是 尚未足够实现这些目标。 从大肠杆菌中选择了来自大肠杆菌的TRP抑制器作为详细的模型系统 研究，因为它是最小，最简单的完整监管之一 今天可用的系统。从已知的结构开始 抑制剂蛋白，带有和没有激活剂L-色氨酸和 三元DNA-Repressor-L-Trp复合物，有可能系统地 研究每个结构元件（氨基酸残基）在 （l）与L-TRP配体保持特定接触，（2）与 操作员DNA和（3）传输调节DNA结合的信号 在绑定位点之间。这将通过研究点和 已知会改变构象和动力学的多个突变 系统。已经获得了非常有希望的初始结果 有了一些突变体，为将来的研究提供了明确的方向。 调查结果的相关性应允许对信号的理解 代码和监管设备的设计。