CHARACTERIZATION OF THE B SUBTILIS APASE GENE FAMILY

枯草芽孢杆菌至少有5个APase基因组成的多基因APase家族。一个 APase多基因家族以前只在哺乳动物中观察到。 对芽孢杆菌APase家族的生化和遗传学研究 基于磷酸盐诱导的APase基因亚家族鉴定 饥饿或产孢量诱导，或两者兼而有之。 这项提议的长期目标是理解生理上的 APase基因家族中每个成员的功能。事实是， 枯草杆菌的各种APase位于细胞内的不同位置， 在开发过程中的不同时间产生，并且具有不同的 比活力和底物专一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极好的模型 系统，结合强大的遗传学和发育系统来评估 磷酸酶的作用。 正在分析每个APase亚家族的原型APase，以确定： 1)表达的调控机制；2)表达的最终归宿 成熟蛋白；3)APase的蛋白质结构。这项规定 研究将确定哪种细胞类型(产孢子或营养)的APase 具有职能作用。 定位数据将指示母细胞或前孔中的角色 如果APase在孢子形成细胞或胞外或 如果APase在营养磷酸盐中表达，在细胞内的作用- 饥饿的细胞。 对APase结构的研究将使实验设计能够确定 蛋白质的表面域是否在蛋白质中起作用 定位或蛋白质-蛋白质相互作用，这些因素可能决定 底物专一性和生理功能。 另一个长期目标是表征pho调节子的组成部分。 在芽孢杆菌中。这些信息对于理解 APase基因家族，并确定pho调节子或PI浓度 在产孢子过程中起作用。

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.