E COLI MIN PROTEINS

大肠杆菌低蛋白

• 批准号: 6351346
• 负责人: LAWRENCE I ROTHFIELD
• 金额: $ 24.49万
• 依托单位: UNIVERSITY OF CONNECTICUT SCH OF MED/DNT
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-02-01 至 2004-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6351346
• 关键词: Escherichia coli; bacterial genetics; bacterial proteins; cell cycle; fluorescence microscopy; gene mutation; genetic mapping; protein localization; protein structure

The long-range goal of this project is to determine the mechanism used by bacteria to select the proper division site at midcell in preference to other potential division sites that are located elsewhere within the cell. Since the products of the three min genes, minC, minD and minE, are responsible for this site-selection process, we will attempt to define the mechanism of action of each of the gene products in the site-selection process. Previous work has shown that MinD localizes to potential division sites at poles and midcell in the absence of the other Min proteins, and that coexpression of MinE or domains of MinE causes redistribution of membrane-associated Gfp-MinD into structures whose position depends on the presence of the MinE topological specificity domain. The major immediate aims of the proposal will be: i. To define the determinants in MinD that are responsible for its localization to poles and midcell, its role in formation of the MinE ring, and its ability to activate the MinC division inhibitor; ii. To determine the molecular basis of the different membrane-associated MinD structures that are induced by coexpression of Gfp-MinD with MinE and with the N-terminal region of MinE, using a combination of fluorescence microscopy, molecular genetics, and membrane-biochemistry; iii. To determine the localization pattern of MinC and the effects of MinC on localization of MinD and MinE; iv. To complete the high-resolution three-dimensional structure of the MinE topological specificity domain and to map topological specificity mutations of MinE to the three-dimensional structure; v. To characterize new minicell mutants that map outside of the minCDE locus and determine whether any of them code for the topological targets for MinD and MinE localization.

该项目的远距离目标是确定细菌用来选择Midcell的适当分裂位点的机制，而不是其他潜在的分裂位点，这些分裂位点位于细胞中的其他地方。 由于三分钟基因的产物Minc，Mind和Mine是负责此场地选择过程的原因，因此我们将尝试在场地选择过程中定义每个基因产物的作用机理。 先前的工作表明，在没有其他最小蛋白质的情况下，思维定位于极点和中部电池的潜在分裂位点，而矿山或矿山的共表达会导致膜相关的GFP的重新分布到其位置，其位置取决于地雷拓扑特异性域的存在。 该提案的主要目的是：i。为了定义负责其定位到两极和中间细胞的决定因素，其在矿环形成中的作用以及激活MINC分裂抑制剂的能力； ii。通过使用荧光显微镜，分子遗传学和膜生物化学的组合，通过与矿山和矿山的N末端区域共表达不同膜相关的思维结构的分子基础； iii。确定MINC的本地化模式以及MINC对思想和矿山定位的影响； iv。要完成矿山拓扑特异性域的高分辨率三维结构，并将我的拓扑特异性突变映射到三维结构； v。要表征新的微型突变体，这些突变体在Mincde基因座之外绘制映射，并确定是否有任何一个为思想和地雷定位的拓扑目标编码。