Studies of E. coli Min proteins

大肠杆菌 Min 蛋白的研究

• 批准号: 7039043
• 负责人: LAWRENCE I ROTHFIELD
• 金额: $ 35.4万
• 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 2009-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7039043
• 关键词: Escherichia coli; bacterial genetics; bacterial pigment; bacterial proteins; biochemistry; cell cycle; chromosome movement; cytoskeleton; fluorescence microscopy; gene mutation; protein localization; protein protein interaction; protein structure; protein structure function

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.

描述（由申请人提供）：该项目的长期目标是确定细菌在中细胞中选择适当细胞分裂位点的机制。三种Min蛋白，MinC， MinD和MinE，是正确选择位点所必需的。蛋白质具有独特的细胞定位模式，并经历独特的极对极振荡周期。我们最近表明，MinCDE蛋白被组织成螺旋状细丝，在细胞两极之间缠绕在细胞骨架状结构中，这种结构与MreB蛋白形成的卷曲结构截然不同。在拟议的资助期间，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：1 .最小蛋白质结构和功能-确定干扰MinCDE系统拓扑特异性功能的突变的影响；Min系统的远程组织-定义细胞内MinDE细胞骨架样元件的组织，确定Min和MreB螺旋丝是否为独立结构，表征与MinD和MinE相互作用的蛋白质，并确定Min螺旋丝与染色体分离之间是否存在关系。MreB-分离MreB和Min细胞骨架样结构，表征与MreB相互作用的蛋白质，并定义MreB细胞骨架样结构在细胞周期过程中的行为。