RECONSTITUTION OF PILUS BIOGENESIS IN A CELL-FREE SYSTEM

致病菌入侵的第一步是附着 它的主人。附着通常由毛发状的附属物介导， 从细菌表面放射出来的，称为皮利。了解菌毛 组装将提供一种重要的防御细菌的途径， 相关疾病，并将提供洞察的基本 细胞器生物发生的生物学过程。这项建议旨在 在无细胞系统中重建菌毛生物发生， 件. P皮利从尿路致病性大肠杆菌，一个充分研究 模型菌毛将用于复溶。P皮利是复杂的 结构由至少六个不同的亚基组成，需要两个 专门的装配蛋白：外膜引导和周质 监护人外膜引导器将并入脂质体中， 菌毛亚单位会被困在里面或被添加到外面 这些蛋白脂质体。分子伴侣-亚基复合物与 usher蛋白，菌毛亚结构的组装，以及完整的 检查皮利。菌毛生物发生的成功重建将 为分析突变体和确定 亚基和组装蛋白的功能。重组将 有助于理解菌毛与宿主细胞的相互作用， 有助于抗菌剂的设计和测试。

The first step in the invasion of a pathogenic bacterium is attachment to its host. Attachment is often mediated by hairlike appendages that radiate out from the bacterial surface, called pili. Understanding pilus assembly will provide an important avenue of defense against bacterial- related diseases, and will also provide insight into the fundamental biological process of organelle biogenesis. This proposal aims to reconstitute pilus biogenesis in a cell-free system using defined components. P pili from uropathogenic Escherichia coli, a well studied model pilus, will be used for reconstitution. P pili are complex structures composed of at least six different subunits, and require two specialized assembly proteins: an outer membrane usher and a periplasmic chaperone. The outer membrane usher will be incorporated into liposomes, and pilus subunits will then be trapped inside or added to the outside of these proteoliposomes. Binding of chaperone-subunit complexes to the usher protein, assembly of pilus substructures, and assembly of complete pili will be examined. Successful reconstitution of pilus biogenesis will provide a powerful tool for analyzing mutants and determining the function of the subunits and assembly proteins. Reconstitution will facilitate the understanding of pilus-host cell interactions, and will aid in the design and testing of antimicrobial agents.