ABRB BINDING TO DEVELOPMENTALLY CONTROLLED PROMOTERS

ABRB 与发育控制启动子的结合

• 批准号: 2184197
• 负责人: MARK A STRAUCH
• 金额: $ 19.28万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 1992
• 资助国家: 美国
• 起止时间: 1992-02-01 至 1997-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2184197
• 关键词: Bacillus subtilis; DNA binding protein; DNA directed RNA polymerase; DNA footprinting; X ray crystallography; bacterial proteins; binding proteins; chemical fingerprinting; cilium /flagellum motility; conformation; developmental genetics; gene deletion mutation; gene expression; genetic promoter element; microorganism culture; mutant; protein structure function; regulatory gene; transcription factor

The AbrB protein of Bacillus subtilis regulates a number of genes, including some essential for the sporulation process, that are expressed during the transition state between balanced exponential growth and stationary phase. It can act as either a positive or negative regulator of transcription, depending upon the particular target gene. It is a DNA-binding protein with specificity toward the promoters it controls. The determinants of this specificity are largely unknown but appear to be related to some type of three-dimensional DNA structure that can be assumed by a variety of different base sequences. We propose to examine the AbrB-DNA interaction in greater detail with the goal to understand more fully how AbrB discriminates between the promoters it controls and those it does not. These studies will include obtaining detailed In vitro footprints of AbrB binding to DNA and correlating this information to the intracellular situation by the use of in vivo footprinting techniques where possible. We will also isolate mutant binding sites and construct in vitro deletion and insertion variants to further delimit elements necessary for recognition. Second site mutants affecting AbrB activity will be searched for in order to more fully understand any other factors that impinge upon AbrB-mediated regulation. Where possible, we will assay the effects that other regulatory proteins may have upon AbrB activity. As one possible mechanism for AbrB-mediated regulation is through its effect upon the localized structure of the promoter, we will examine the effects of AbrB binding upon changes in DNA bending. We will also initiate a crystallographic analysis of the AbrB protein with the ultimate goal of relating structure to function.

枯草芽孢杆菌的AbrB蛋白调节许多基因， 包括一些对孢子形成过程至关重要的物质， 在平衡指数增长和 固定相 它可以作为一个积极或消极的调节剂， 这取决于特定的靶基因。 这是一个 对它所控制的启动子具有特异性的DNA结合蛋白。 的 这种特异性的决定因素在很大程度上是未知的，但似乎是 与某种三维DNA结构有关， 由不同的碱基序列组成 我们建议研究 AbrB-DNA相互作用的更详细的目标，以了解更多 AbrB如何充分区分它控制的启动子和那些它控制的启动子， 并没有。 这些研究将包括获得详细的体外 AbrB与DNA结合的足迹，并将此信息与 细胞内的情况，通过使用体内足迹技术，其中 可能 我们还将分离突变体结合位点并在体外构建 删除和插入变体，以进一步界定 识别. 将搜索影响AbrB活性的第二位点突变体 因为为了更全面地了解任何其他影响 AbrB介导的调节。 在可能的情况下，我们将分析 其它调节蛋白可能对AbrB具有活性。 作为一个可能 AbrB介导的调节机制是通过其对 启动子的局部结构，我们将检查AbrB的影响， 结合在DNA弯曲的变化上。 我们还将发起一项 AbrB蛋白的晶体学分析，最终目标是 将结构与功能联系起来。