Genetic Competence Apparatus of Bacillus Subtilis

枯草芽孢杆菌遗传能力仪

• 批准号: 7447970
• 负责人: DAVID A DUBNAU
• 金额: $ 19.12万
• 依托单位: UNIV OF MED/DENT OF NJ-NJ MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 1990
• 资助国家: 美国
• 起止时间: 1990-12-01 至 2007-12-18
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7447970
• 关键词: ATP phosphohydrolase; Bacillus subtilis; Bacteria; Binding; Binding Proteins; Biochemical; Biochemical Genetics; Cell membrane; Cell surface; Cells; Competence; Complex; DNA; DNA Binding; Disulfides; Endopeptidases; Genes; Genetic; Genetic Transcription; Helix (Snails); In Vitro; Individual; Ion Channel; Localized; Membrane; Molecular; Open Reading Frames; Oxidation-Reduction; Peptide Hydrolases; Physiological; Process; Property; Proteins; Role; Structure; Sulfhydryl Compounds; System; Thiol Disulfide Oxidoreductase; Transport Process; Vesicle; Work; base; disulfide bond; ds-DNA; ear helix; mutant; protein folding; receptor; uptake

Natural transformation in bacteria refers to the uptake and integration of exogenous DNA. Bacillus subtilis can be transformed by exogenous DNA when in a physiological state known as competence. Competent cells are able to bind double stranded DNA, fragment the DNA on the cell surface and transport a single strand across the cell membrane. Our long-term objective is to understand these processes on the molecular evel. This proposal explores the specific roles of proteins known to participate in the binding, processing and transport (internalization) of transforming DNA. Several of these proteins (the seven comG gene products, ComC and ComEA) are needed for the binding of DNA to the cell surface while others (ComEC, ComFA and ComEA) are required for transport of DNA across the cell membrane. The proposed work will explore interactions among these proteins and the role of the thiol-disulfide oxidoreductases BdbD and BdbC in the folding of these proteins and in the assembly of the DNA uptake apparatus. The formation and properties of a newly identified higher order ComGC complex will be elucidated and its role in DNA binding will be examined. The structure of the putative membrane channel protein ComEC and the in vitro biochemical activities of the transporter ATPase ComFA will be characterized further. The amounts of these various proteins in the cell will be determined. Several newly identifed transformation proteins will be studied. Finally an attempt will be made to develop a vesicle-based in vitro DNA transport system.

细菌的自然转化是指外源DNA的摄取和整合。枯草芽孢杆菌 当处于称为能力的生理状态时，可以被外源DNA转化。胜任的 细胞能够结合双链 DNA，将细胞表面的 DNA 片段化并运输单链 DNA。 链穿过细胞膜。我们的长期目标是了解这些过程 分子水平。该提案探讨了已知参与结合的蛋白质的具体作用， 转化 DNA 的加工和运输（内化）。其中一些蛋白质（七个 DNA 与细胞表面的结合需要 comG 基因产物（ComC 和 ComEA），而 DNA 跨细胞膜运输需要其他物质（ComEC、ComFA 和 ComEA）。这 拟议的工作将探索这些蛋白质之间的相互作用以及硫醇二硫化物的作用 氧化还原酶 BdbD 和 BdbC 参与这些蛋白质的折叠和 DNA 摄取的组装 设备。新鉴定的高阶 ComGC 复合物的形成和性质将是 阐明并检查其在 DNA 结合中的作用。假定的膜通道的结构 蛋白质 ComEC 和转运蛋白 ATPase ComFA 的体外生化活性将 进一步表征。细胞中这些不同蛋白质的含量将被测定。几款新 将研究已鉴定的转化蛋白。最后将尝试开发一个 基于囊泡的体外 DNA 运输系统。