SITE-SPECIFIC ENERGETICS OF PROTEIN-DNA INTERACTIONS

蛋白质-DNA 相互作用的位点特异性能量

• 批准号: 3296230
• 负责人: Gary K Ackers
• 金额: $ 12.47万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1989
• 资助国家: 美国
• 起止时间: 1989-07-01 至 1994-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3296230
• 关键词: DNA; DNA footprinting; acidity /alkalinity; autoradiography; bacteriophage lambda; bioenergetics; chemical binding; gel filtration chromatography; genetic regulation; image processing; ionic strengths; mathematical model; mutant; proteins; temperature; thermodynamics; transcription factor

The goal of this research program is to elucidate the physical mechanism of genetic control processes mediated by the interactions of proteins at specific sites on DNA. Components of the program include: (a) development of new techniques for the study of site- specific protein-DNA interactions, (b) theoretical work on the thermodynamic of coupled individual-site binding reactions in multi-site gene control systems, (c) experimental studies of functional energetics in specific gene control systems, (d) correlation of the energetic properties with structural features of the interacting molecules, (e) development and testing of physical-chemical models for gene control systems. We plan to extend our previous studies on the repressor-operator control system of bacteriophage lambda, which is a prototype for a large family of cooperative multi-site gene control systems. Using the Quantitative DNase Footprint Titration technique that we have recently developed, we will carry out a detailed study of the effects of temperature, pH, and ionic interactions on the site- specific energetics (including cooperativity effects) of cI and cro repressor binding to the DNA operator sites. The results will provide new insights into the detailed roles of proton binding, cation binding, and the various non-covalent forces responsible for the regulatory interactions. We will study the biological role of cooperative interactions between operator-bound repressors by in vivo and in vitro studies of mutant repressors that have altered cooperativity, using a physical-chemical model of gene control. We also plan further development of theory and techniques for understanding site-specific protein-DNA interactions.

该研究计划的目标是阐明身体 相互作用介导的遗传控制过程机理 DNA上特定位点的蛋白质的蛋白质。 程序的组成部分 包括：（a）开发新技术以研究站点 - 特定的蛋白质-DNA相互作用，（b）关于 耦合的单位结合反应的热力学 多站点基因控制系统，（c）实验研究 特定基因控制系统中的功能能力，（d） 能量特性与结构特征的相关性 相互作用的分子，（e）开发和测试 基因控制系统的物理化学模型。 我们计划扩展以前的有关阻遏操作员的研究 噬菌体lambda的控制系统，这是一个原型 一个合作多站点基因控制系统的大家族。 使用我们的定量DNase足迹滴定技术 最近开发了，我们将对 温度，pH和离子相互作用对位点的影响 CI和CRO的特定能量学（包括协同效应） 阻遏物与DNA操作员位点结合。结果将 提供有关质子结合的详细作用的新见解， 阳离子结合以及负责的各种非共价力 监管相互作用。 我们将研究合作互动的生物学作用 在体内和体外研究之间结合操作员的阻遏物之间 使用A的突变阻遏物的使用 基因控制的物理化学模型。 我们还计划进一步发展理论和技术 了解位点特异性蛋白-DNA相互作用。