Molecular Mechanisms of the Type I Secretion System from Bacterial Pathogens

细菌病原体I型分泌系统的分子机制

基本信息

批准号：
10795448
负责人：
Wei Mi
金额：
$ 6.35万
依托单位：
YALE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-08-01 至 2026-07-31
项目状态：
未结题

项目摘要

ABSTRACT Protein secretion is an essential component of the arsenal used by many bacterial pathogens to invade hosts, damage tissues, and suppress immune responses. Gram-negative bacteria have evolved sophisticated machines to secrete proteins across two membranes: the inner membrane and outer membrane. Since discovered in 1979, the type I secretion system (T1SS) has been identified in more than 800 bacterial genomes, and many of its substrates contribute to the virulence of pathogens, examples being hemolysin HlyA in uropathogenic Escherichia coli, adhesin SiiE in Salmonella enterica, and adenylate cyclase CyaA in Bordetella pertussis. The T1SS secretes unfolded substrates from the cytoplasm to the extracellular milieu. It includes three components: an ATP-binding cassette transporter (ABC transporter), associated periplasmic adapter proteins located in the inner membrane, and a porin in the outer membrane. The canonical “alternating access” model cannot explain how the large protein substrates are transported by the T1SS ABC transporter across the inner membrane. Therefore, the mechanisms of substrate secretion at the molecular level is still obscure. The proposed research leverages single particle cryo-electron microscopy (cryoEM), bioinformatics, in vivo and in vitro assays to answer key questions in the field, including the structural basis of substrate recognition, the nature of the translocation pathway, the energetics for substrate translocation, and the regulation of the T1SS ABC transporter's activity. The studies will substantially increase the fundamental scientific knowledge about the mechanisms of the T1SS and provide a new basis for developing future therapeutic interventions against a broad range of bacterial infections.

抽象的蛋白质分泌是许多细菌病原体使用的武器库的重要组成部分侵入宿主、损害组织并抑制革兰氏阴性细菌的免疫反应。跨两个膜（内膜）分泌蛋白质的复杂机器自 1979 年发现以来，I 型分泌系统 (T1SS) 已被广泛研究。在 800 多个细菌基因组中发现，其许多底物有助于病原体的毒力，例如尿路致病性大肠杆菌中的溶血素 HlyA，肠沙门氏菌中的粘附素 SiiE 和百日咳博德特氏菌中的腺苷酸环化酶 CyaA。 T1SS 将未折叠的底物从细胞质分泌到细胞外环境，它包括三种。成分： ATP 结合盒转运蛋白（ABC 转运蛋白）、相关周质接头蛋白位于内膜，孔蛋白位于外膜。规范的“交替访问”模型无法解释大蛋白质底物是如何存在的由 T1SS ABC 转运蛋白跨内膜转运。分子水平上的底物分泌机制仍然不清楚。利用单粒子冷冻电子显微镜 (cryoEM)、生物信息学、体内和体外回答该领域关键问题的测定，包括底物识别的结构基础，易位途径的性质、底物易位的能量学以及 T1SS ABC 转运蛋白活性的调节这些研究将大大增加关于 T1SS 机制的基础科学知识并提供新的基础开发针对广泛细菌感染的未来治疗干预措施。