THE PENICILLIN-BINDING PROTEINS OF BACILLUS SUBTILIS

本项目的目的是检查和评估 不同培养条件下枯草芽孢杆菌青霉素结合蛋白的研究 生理条件。长期目标是将 这些蛋白质的活性与生长等生物学过程有关 细胞壁，在细胞分裂或产孢期形成隔膜， 发育中的前孔皮质层合成。因为所有的 据推测，这些事件容易受到青霉素的抑制。 至少部分项目管理方的行动必须对他们的 正常完成。将作出重大努力来评估PBP的变化 发生在产孢期，因为有两种青霉素敏感 在这个发展过程中的几个步骤，因为早期的 孢子形成与营养细胞中发生的事件非常相似 组织。PBP波动的性质，它们的物理位置， 他们的时机，以及他们的普遍性将被审查。作为对…的补充 在这些研究中，植物多氯联苯的地形分布 细胞膜的确定将通过比较来自 小细胞(主要是细胞末端)和那些来自杆状细胞(主要是 牢房侧壁)。此外，2个菌株的表型特征 现有的PBP缺陷突变体将被用来寻找一些生物学上的 可归因于特定PBP缺陷的属性或功能。 这些PBP突变体和其他可能在我们建议的调查中发现的 枯草杆菌现有的细胞形态和细胞分裂突变体将是 基因分析的对象开始绘制PBP基因图谱， 确定它们之间的联系，并将它们与特定的 表型特征。这里提出的各种方法 应该大大增加我们对体内角色的稀疏知识 这些多氯联苯。这些信息将补充和扩展 我们的理解主要来自体外生化分析。 这些结果应该会为我们提供一些重要和有用的见解 细胞分裂和孢子形成的机制、它们的调节和它们的 青霉素抑制作用。在另一个层面上，关于以下方面的附加信息 产孢子最终使我们能够更好地操纵这一过程 生产各种产孢量特定的产品，如蛋白水解物 工业和医疗用的酶、外毒素和抗生素。

The purpose of this project is to examine and evaluate the penicillin-binding proteins (PBPs) of Bacillus subtilis under a variety of physiological conditions. The long range goal is to correlate the activities of these proteins with such biological processes as growth of the cell wall, formation of the septum during cell division or sporulation, and synthesis of the cortex layer in the developing forespore. Since all these events are susceptible to inhibition by penicillin, it is assumed that the action of at least some of the PBPs must be essential for their normal completion. A major effort will be made to evaluate the PBP changes that occur during sporulation because there are 2 penicillin-sensitive steps in this developmental process and because the early stages of sporulation are very similar to events occuring in a vegetative cell division. The nature of the PBP fluctuations, their physical location, their timing, and their universality will be examined. As a complement to these studies, the topographical distribution of the vegetative PBPs within the cell membrane will be determined by a comparison of membranes from minicells (primarily cell ends) with those from rod-shaped cells (primarily cell sidewalls). Furthermore, a phenotypic characterization of the 2 existing PBP-deficient mutants will be done to look for some biological property or function that can be attributed to the specific PBP defect. These PBP mutants and others that may be found in our proposed survey of the available cell shape and cell division mutants of B. subtilis will be the subjects of genetic analysis to begin to map the PBP genes, to determine their linkage to one another, and to correlate them with specific phenotypic properties. The various approaches that are proposed here should significantly add to our sparse knowledge of the in vivo roles of these PBPs. This information will both complement and extend the understanding we have gained primarily from in vitro biochemical analyses. The results should provide some important and useful insights into the mechanisms of cell division and sporulation, their regulation, and their inhibition by penicillin. On another level, additional information about sporulation could ultimately enable us to manipulate the process better for the production of various sporulation-specific products such as proteolytic enzymes, exotoxins, and antibiotics for industrial and medical uses.