QUANTITATIVE STUDIES OF THE BIOTIN REGULATORY SYSTEM

生物素调节系统的定量研究

• 批准号: 2184010
• 负责人: DOROTHY BECKETT
• 金额: $ 9.39万
• 依托单位: UNIVERSITY OF MARYLAND BALTIMORE COUNTY
• 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 1997-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2184010
• 关键词: DNA binding protein; DNA footprinting; Escherichia coli; SDS polyacrylamide gel electrophoresis; biological models; biophysics; biotin; chemical binding; chimeric proteins; dimer; fluorescence spectrometry; gene induction /repression; genetic operator element; genetic regulation; genetic transcription; intermolecular interaction; ionic strengths; monomer; mutant; nutrition related tag; protonation; stoichiometry; temperature; thermodynamics; transcription factor; transport proteins; vitamin biosynthesis

Regulation of gene expression in prokaryotes and eukaryotes is effected by complex macromolecular assemblies. Function of these assemblies can depend on protein-protein, protein-small molecule, and protein-DNA interactions. Regulation of biotin biosynthesis and retention in E. coli depends on all three types of interactions. A model for the biotin system, based on a qualitative understanding of the macromolecular processes that contribute to the regulation, has been proposed. The biotin system provides an ideal model system for relating the physical chemistry of individual macromolecular interactions to the biology of a complex gene regulatory system. The goals of this research proposal are to: 1. perform quantitative studies of the interactions that contribute to biotin regulation to define the functional energetics of the biotin system. 2. investigate the thermodynamic driving forces responsible for the regulatory interactions. 3. evaluate and revise the qualitative model for biotin regulation in light of the results of the physical studies and develop and test quantitative models for the system. The techniques that will be employed in these studies include a. the quantitative DNAse footprint technique to measure DNA binding, b. equilibrium sedimentation to determine of subunit assembly properties, and c. fluorescence spectroscopy to measure heterologous protein-protein interactions. These studies of the biotin system will help define the relationship between the physical chemistry of individual biotin regulatory interactions and the biology of the entire system and may reveal general principles that are relevant to understanding more complex gene regulatory systems. Thermodynamic characterization of the interactions will, moreover, yield information about the physical chemical forces that drive these interactions.

原核生物和真核生物中基因表达的调节是 由复杂的大分子组件产生。 这些功能 组件可以取决于蛋白质蛋白质，蛋白质 - 小分子， 和蛋白-DNA相互作用。 生物素生物合成的调节 大肠杆菌中的保留率取决于所有三种类型的 互动。 生物素系统的模型，基于 对大分子过程的定性理解 已提出有助于该法规。 生物素系统 提供了关联物理化学的理想模型系统 单个大分子相互作用与生物学的生物学 复杂的基因调节系统。 这项研究的目标 提案是： 1。对贡献的相互作用进行定量研究 为了定义生物素调节以定义 生物素系统。 2。研究负责的热力学驱动力 监管相互作用。 3。评估和修改生物素调节的定性模型 鉴于物理研究的结果并发展和 测试系统的定量模型。 这些研究将采用的技术包括a。 测量DNA结合的定量DNase足迹技术， b。平衡沉积以确定亚基组件 属性和c。荧光光谱以测量 异源蛋白质蛋白相互作用。 这些研究 生物素系统将有助于定义 单个生物素调节性相互作用的物理化学和 整个系统的生物学，可能揭示一般原理 与理解更复杂的基因调节有关的 系统。 相互作用的热力学表征将 此外，产生有关物理化学力的信息 驱动这些互动。