STUDIES OF BACTERIAL MINICELL MUTANTS

细菌小细胞突变体的研究

• 批准号: 3300501
• 负责人: LAWRENCE I ROTHFIELD
• 金额: $ 23.68万
• 依托单位: UNIVERSITY OF CONNECTICUT SCH OF MED/DNT
• 依托单位国家: 美国
• 财政年份: 1989
• 资助国家: 美国
• 起止时间: 1989-04-01 至 1996-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3300501
• 关键词: DNA replication; Escherichia coli; adenosinetriphosphatase; bacterial genetics; bacterial proteins; cell cycle; crosslink; gene deletion mutation; gene expression; immunocytochemistry; immunoelectron microscopy; laboratory rabbit; microorganism growth; operon; point mutation; suppressor mutations; temperature sensitive mutant

The long-range objective of this project is to understand the mechanism used by bacterial cells to place the division site at its proper location at midcell. To approach this objective, the bacterial minicell locus (minB) will be studies since we have previously shown that the three min gene products play an important role during the normal cell cycle to ensure the proper placement of the division septum at midcell. This is accomplished by the combined action of a division inhibitor, dependent on the presence of the MinC and MinD proteins, and a topological specificity factor, the MinE protein. A combination of genetics, immunochemistry and immunoelectron microscopy, and biochemistry will be used, with the following goals: (1) to determine the cellular localization of the three Min proteins under normal conditions and under conditions where the topological specificity of septal placement is perturbed; (2) to determine which proteins interact with the Min proteins within the cell; (3) to identify the domain within MinE that is responsible for its ability to give topological specificity to the site-selection process, and to use genetic means to identify cellular proteins that play a role in the topological specificity function of MinE; (4) to use genetic means to identify cellular proteins that play a role in the MinC-mediated septation block; (5) to determine how the ATPase activity of MinC- mediated septation block; (5) to determine how the ATPase activity of MinD is coupled to its role in the site-selection process.

该项目的长期目标是了解该机制 细菌细胞利用其将分裂位点放置在适当的位置 在中单元。 为了实现这一目标，细菌小细胞位点 （minB）将被研究，因为我们之前已经表明，三分钟 基因产物在正常细胞周期中发挥重要作用 确保分裂隔膜正确放置在中室。 这是 通过分裂抑制剂的联合作用来完成，取决于 MinC 和 MinD 蛋白的存在以及拓扑特异性 因子，MinE 蛋白。 遗传学、免疫化学和 将使用免疫电子显微镜和生物化学， 目标如下：（1）确定三者的细胞定位 正常条件下和条件下的最小蛋白质 间隔位置的拓扑特异性受到干扰； (2) 至 确定哪些蛋白质与细胞内的 Min 蛋白质相互作用； (3) 识别 MinE 内负责其的域 能够为选址过程提供拓扑特异性， 并使用遗传手段来识别发挥作用的细胞蛋白质 MinE 的拓扑特异性函数； (4)利用遗传手段 鉴定在 MinC 介导的细胞内发挥作用的细胞蛋白 分隔块； (5) 测定MinC-的ATPase活性如何 介导的分隔阻滞； (5) ATP酶活性的测定 MinD 与其在选址过程中的作用相结合。