Quantitative Studies of the Biotin Regulatory System

生物素调节系统的定量研究

• 批准号: 6519465
• 负责人: DOROTHY BECKETT
• 金额: $ 23.22万
• 依托单位: UNIV OF MARYLAND, COLLEGE PARK
• 依托单位国家: 美国
• 财政年份: 1992
• 资助国家: 美国
• 起止时间: 1992-05-01 至 2005-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6519465
• 关键词: DNA footprinting; Escherichia coli; X ray crystallography; bacterial genetics; bacterial proteins; biotin; calorimetry; gene induction /repression; microorganism culture; operon; protein protein interaction; site directed mutagenesis; transcription factor; vitamin biosynthesis

DESCRIPTION (provided by applicant): Protein-protein interactions are critical to regulation of expression of genetic information. The strength of homologous protein-protein interactions dictates the level of occupancy of transcriptional regulatory sites on DNA. Patterns of heterologous protein-protein interactions determine the selection of DNA target site and/or the level of occupancy of a site. Through their influence on DNA binding these macromolecular interactions are fundamental to control of expression of genetic information. Elucidation of the details necessary for successful clinical intervention at the level of protein-protein interactions in control of gene expression requires combined solution physico-chemical studies of function as well as high-resolution structural information. The Escherichia coli biotin regulatory system provides an excellent model system for detailed studies of the control of a genetic regulatory switch via protein-protein interactions. In this proposal experiments designed to determine the structural and thermodynamic features of corepressor-induced assembly of a transcriptional repressor are described. Strategies to examine the control of gene expression via heterologous protein-protein interactions are also outlined. Methods utilized in the proposed work include site-directed and random mutagenesis, stopped-flow fluorescence kinetic measurements, sedimentation equilibrium, DNaseI footprinting, isothermal titration calorimetry and x-ray crystallography. Results of these combined studies will yield information of general significance to understanding the detailed chemistry of the protein-protein interactions essential to regulation of genetic expression.

描述（由申请人提供）：蛋白质-蛋白质相互作用至关重要 来调节遗传信息的表达。同源实力 蛋白质-蛋白质相互作用决定转录占据的水平 DNA 上的调控位点。异源蛋白质-蛋白质相互作用的模式 确定DNA靶位点的选择和/或a的占据水平 地点。通过它们对 DNA 结合的影响，这些大分子相互作用 是控制遗传信息表达的基础。阐明 成功临床干预所需的细节 控制基因表达的蛋白质-蛋白质相互作用需要结合 功能的溶液物理化学研究以及高分辨率 结构信息。大肠杆菌生物素调节系统提供 一个优秀的模型系统，用于详细研究遗传控制 通过蛋白质-蛋白质相互作用进行调节开关。在这个提案中 旨在确定结构和热力学特征的实验 描述了辅阻遏物诱导的转录阻遏物的组装。 通过异源检查基因表达控制的策略 还概述了蛋白质-蛋白质相互作用。中使用的方法 拟议的工作包括定点突变和随机突变、停流 荧光动力学测量、沉降平衡、DNaseI 足迹法、等温滴定量热法和 X 射线晶体学。 这些综合研究的结果将产生一般信息 对于理解蛋白质-蛋白质的详细化学性质具有重要意义 基因表达调控所必需的相互作用。