Studies of E. coli Min proteins

该项目的长期目标是确定细菌选择合适细胞的机制 分裂位点在中细胞。三种Min蛋白，MinC，MinD和MinE，是合适位点所必需的 选择.这些蛋白质具有独特的细胞定位模式， 振荡周期我们最近发现MinCDE蛋白被组织成螺旋丝， 在两极之间缠绕在细胞周围，形成类似细胞器的结构， 由MreB蛋白形成的结构。在建议的资助期内，我们将使用以下组合： 荧光显微镜，生物化学和遗传学，以实现以下目标：i.最小蛋白质结构 和功能-为了确定干扰拓扑特异性功能的突变的影响， MinCDE系统，ii.民系统的远程组织--定义民系统的组织 MinDE细胞内的胞浆样分子，以确定Min和MreB是否为螺旋丝 是独立的结构，以表征与MinD和MinE相互作用的蛋白质，并确定是否 Min螺旋丝与染色体分离之间存在关系，iii. MreB--分离 MreB和Min细胞间隙样结构，以表征与MreB相互作用的蛋白质，并定义 在细胞周期过程中MreB细胞间隙样结构的行为。

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.