STUDIES OF BACTERIAL MINICELL MUTANTS

本项目的长期目标是了解 细菌细胞用来将分裂位点定位在适当的位置 在midcell。 为了实现这一目标，细菌小细胞基因座 （minB）将是研究，因为我们以前已经表明，三个min 基因产物在正常细胞周期中起重要作用， 确保分裂隔在中间细胞的正确位置。 这是 通过分裂抑制剂的联合作用完成，取决于 MinC和MinD蛋白的存在，以及拓扑特异性 MinE蛋白。 遗传学、免疫化学和 免疫电镜和生物化学将被使用， 以下目标：（1）确定三个细胞的定位 在正常条件下以及在 间隔位置的拓扑特异性受到干扰;（2） 确定哪些蛋白质与细胞内的Min蛋白质相互作用; (3)确定MinE中负责其 为位点选择过程提供拓扑特异性的能力， 并利用遗传学手段来鉴定细胞蛋白质， 在MinE的拓扑特异性功能中的作用;（4）利用遗传学手段 为了鉴定在MinC介导的 （5）测定MinC-ATP酶活性 介导的分隔阻滞;（5）测定ATP酶活性， MinD与其在选址过程中的作用相关联。

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.