Studies of E. coli Min proteins

描述(申请人提供)：这个项目的长期目标是确定细菌用来选择合适的中细胞分裂部位的机制。Minc、Mind和My这三种Min蛋白是选择合适的位点所必需的。这些蛋白质具有独特的细胞定位模式，并经历独特的极点到极点振荡周期。我们最近发现，MinCDE蛋白被组织成螺旋丝，缠绕在细胞两极之间的细胞骨架样结构中，似乎不同于MreB蛋白形成的卷曲结构。在拟议的赠款期间，我们将使用荧光显微镜、生物化学和遗传学的组合来实现以下目标：i.Min蛋白质结构和功能--确定干扰MinCDE系统的拓扑特异性功能的突变的影响，i.Min系统的长期组织--确定细胞内Minde类细胞骨架元件的组织，确定Min和MreB螺旋细丝是否是独立的结构，确定与Mind和MreB相互作用的蛋白质的特征，并确定Min螺旋细丝和染色体分离之间是否存在关系，3.MreB--分离MreB和Min类细胞骨架结构，鉴定与MreB相互作用的蛋白质，并确定MreB类细胞骨架结构在细胞周期过程中的行为。

DESCRIPTION (provided by applicant): The long-range goal of this project is to determine the mechanism used by bacteria to select the proper cell division site at midcell. The three Min proteins, MinC, MinD and MinE, are required for proper site selection. The proteins have a unique cellular localization pattern and undergo a unique pole-to-pole oscillatory cycle. We have recently shown that the MinCDE proteins are organized into spiral filaments that wind around the cell between the two poles in a cytoskeletal-like structure that appears distinct from a coiled structure formed by the MreB protein. During the proposed grant period we will use a combination of fluorescence microscopy, biochemistry and genetics to achieve the following aims: i. Min protein structure and function-- To determine the effects of mutations that interfere with the topological specificity function of the MinCDE system, ii. Long-range organization of the Min system-- To define the organization of the MinDE cytoskeletal-like elements within the cell, to determine whether the Min and MreB spiral filaments are separate structures, to characterize proteins that interact with MinD and MinE, and to determine whether there is a relationship between the Min spiral filaments and chromosome segregation, iii. MreB-- To isolate the MreB and Min cytoskeletal-like structures, to characterize proteins that interact with MreB, and to define the behavior of the MreB cytoskeletal-like structures during the course of the cell cycle.