CHARACTERIZATION OF THE B SUBTILIS APASE GENE FAMILY

B。枯草芽孢杆菌具有至少5个APase基因的多基因APase家族。 一个 APase多基因家族以前仅在哺乳动物中观察到。 芽孢杆菌APase家族的生物化学和遗传学研究 基于磷酸盐诱导鉴定APase基因亚家族 饥饿或孢子形成诱导或两者。 这项建议的长期目标是了解生理 APase基因家族的每个成员的功能。 的事实 B的各种AP。枯草芽孢杆菌位于细胞中的不同位置， 在发育过程中的不同时期产生， 特异性活性和底物特异性意味着每种同工酶 在细胞中具有独特的功能。 芽孢杆菌提供了一个很好的模型 系统，将强大的遗传学与发展系统相结合， 磷酸酶的作用 正在分析每个APase亚家族的原型APase，以确定： 1)表达调控机制，2）最终目的地的表达， 成熟蛋白; 3）APase的蛋白结构。 调控 研究将确定在哪种细胞类型（孢子形成或营养）APase 具有功能性作用。 定位数据将指示在母细胞或前孢子中的作用 如果AP酶在孢子形成细胞中表达，或在细胞外或细胞外表达， 细胞内的作用，如果AP酶是在营养磷酸表达- 饥饿的细胞 APase结构的研究将允许设计实验来确定 蛋白质的表面结构域是否在蛋白质 定位或蛋白质-蛋白质相互作用， 底物特异性和生理功能。 另一个长期目标是描述pho调节子的组成 在芽孢杆菌中。 这些信息对于理解以下规则是必要的： APase基因家族，并确定磷酸调节子或Pi浓度 在孢子形成中起作用。

B. subtilis has a multigene APase family of at least 5 APase genes. An APase multigene family has previously been observed only in mammals. Biochemical and genetic studies of the Bacillus APase family have identified APase gene subfamilies based on induction by phosphate starvation or sporulation induction or both. The long-term goal of this proposal is to understand the physiological function of each member of the APase gene family. The fact that the various APases of B. subtilis are located in different places in the cell, are produced at different times during development, and have different specific activities and substrate specificities implies that each isozyme has a unique function in the cell. Bacillus provides an excellent model system, combining powerful genetics with a developmental system to assess the roles of phosphatases. A prototype APase for each APase subfamily is being analyzed to determine: 1) mechanism of regulation of expression, 2) the final destination of the mature protein, and 3) protein structure of the APase. The regulation studies will determine in which cell type (sporulating or vegetative) APase has a functional role. The localization data will indicate a role in the mother cell or forespore if the APase is expressed in the sporulating cell, or an extracellular or intracellular role if the APase is expressed in the vegetative phosphate- starved cell. Study of APase structure will allow design of experiments to determine whether the surface domains of the protein have roles in protein localization or protein-protein interactions, factors that could determine substrate specificity and physiological function. Another long-term goal is to characterize the components of the pho regulon in Bacillus. This information is necessary to understand the regulation of the APase gene family and determine if the pho regulon or Pi concentration has a role in sporulation.