Studies of E. coli Min proteins

这个项目的长期目标是确定细菌用来选择合适细胞的机制 分裂部位在中层细胞。Minc、Mind和My这三种Min蛋白是合适的位点所必需的 选择。这些蛋白质具有独特的细胞定位模式，并经历独特的从一极到另一极的定位 振荡周期。我们最近发现MinCDE蛋白被组织成螺旋状的细丝 在两极之间的细胞周围缠绕，形成类似细胞骨架的结构，看起来与卷曲的结构不同 由MreB蛋白形成的结构。在拟议的授权期内，我们将使用以下组合 利用荧光显微镜、生物化学和遗传学来实现以下目标：i.MIN蛋白质结构 和功能--确定干扰拓扑特异性功能的突变的影响 MinCDE系统，II。MIN系统的远程组织--定义 Minde细胞内的细胞骨架样分子，以确定Min和MreB是否为螺旋细丝 是不同的结构，以表征与Mind和Mine相互作用的蛋白质，并确定 Min螺旋细丝与染色体分离之间存在一定的关系。MreB--分离 MreB和Min细胞骨架样结构，以表征与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。

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, biochemistrry 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.