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.

拟议项目的目标是实现一个详细和准确的 了解遗传调控的分子机制，在一定程度上 这一机制运作中的具体变化可能是 预测和设计。预测和控制修改的能力 基因调控机制的功能将产生深远的影响 对治疗遗传性疾病和使用 机制作为合理药物设计的目标，但现有的知识 但仍不足以实现这些目标。 从E.选择大肠杆菌作为模型系统， 研究，因为它是一个最小和最简单的完整的监管 系统目前可用。从已知的 阻遏蛋白，有和没有激活剂L-色氨酸，以及 三元DNA-阻遏物-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)保持与L-trp配体的特异性接触，和（2）与 操纵子DNA和（3）传递调节DNA结合的信号 在结合位点之间。这将通过研究点和 已知多种突变会改变构象和动力学 该系统已经取得了非常有希望的初步成果 与一些突变体，为未来的研究提供了明确的方向。 这些发现的相关性应该有助于理解 规范和调节装置的设计。