Autotransporter proteins and virulence of Y. pestis

自转运蛋白和鼠疫耶尔森氏菌的毒力

• 批准号: 7017714
• 负责人: VIRGINIA L MILLER
• 金额: $ 29.88万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-03-01 至 2009-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7017714
• 关键词: Escherichia coli; Yersinia pestis; Yersinia pestis disease; antibody; bacterial genetics; bacterial proteins; bioterrorism /chemical warfare; gene deletion mutation; gene expression; histopathology; laboratory mouse; membrane proteins; polymerase chain reaction; protein localization; protein structure function; virulence; western blottings

DESCRIPTION (provided by applicant): Yersinia pestis is a human pathogen that is the causative agent of plague. Unfortunately, due to the high mortality rates and severe disease caused by Y. pestis, Y. pestis has emerged as a potential agent of biological warfare and bioterrorism. Although environmental outbreaks of plague largely have been controlled through the use of modern public health measures, the potential use of this pathogen as an agent of warfare and terror necessitates a more detailed understanding of the pathogenesis of Y. pestis to facilitate the identification of targets for vaccine development and treatment. No vaccine is currently available for either the bubonic or pneumonic form of plague. Examination of the recently published genome of Y. pestis indicates that Y. pestis potentially could express six different autotransporter (AT) proteins (designated Yap for Yersinia autotransporter protein). AT proteins have been identified in a wide variety of pathogens and as their name suggests the primary sequence of the protein is sufficient to direct transport across the outer membrane of the bacterium. In addition, in cases where the function of the AT is known these proteins are uniformly associated with virulence. The presence of six such sequences in the Y. pestis genome is of interest from the standpoint of understanding the interaction of Y. pestis with its host as well as potential vaccine candidates. Thus, to begin to study these ATs of Y. pestis we propose the following specific aims: Aim 1. Analysis of expression and localization of the Yaps in Y. pestis. Six different Yaps are predicted from the genome sequence and they may not all be expressed at the same time or localized in the same manner. A better understanding of the conditions under which each of the yaps is expressed and localized will provide valuable clues as to its potential function. Aim 2. Analysis of the role of Yaps in the virulence of Y. pestis. Mutants of each of the yap genes will be constructed in the CO92 strain of Y. pestis and then tested for virulence using the mouse model of infection. Aim 3. Functional analysis of the Yaps. We will begin to investigate the function of the Yaps by testing E. coli expressing individual Yaps for a variety of known functions previously associated with autotransporter proteins.

描述（由申请人提供）：鼠疫耶尔森氏菌是一种人类病原体，是鼠疫的病原体。不幸的是，由于鼠疫耶尔森氏菌引起的高死亡率和严重疾病，鼠疫耶尔森氏菌已成为生物战和生物恐怖主义的潜在媒介。尽管鼠疫的环境暴发在很大程度上已通过使用现代公共卫生措施得到控制，但这种病原体可能被用作战争和恐怖媒介，因此需要更详细地了解鼠疫耶尔森氏菌的发病机制，以便于确定防治目标。疫苗开发和治疗。目前还没有针对腺鼠疫或肺炎鼠疫的疫苗。对最近发表的鼠疫耶尔森氏菌基因组的检查表明，鼠疫耶尔森氏菌可能表达六种不同的自转运蛋白（AT）（称为耶尔森氏菌自转运蛋白）。 AT 蛋白已在多种病原体中被发现，正如其名称所示，该蛋白的一级序列足以直接转运穿过细菌的外膜。此外，在已知 AT 功能的情况下，这些蛋白质均与毒力相关。从了解鼠疫耶尔森氏菌与其宿主以及潜在候选疫苗相互作用的角度来看，鼠疫耶尔森氏菌基因组中存在六个此类序列是令人感兴趣的。因此，为了开始研究鼠疫耶尔森氏菌的这些 AT，我们提出以下具体目标： 目标 1. 分析鼠疫耶尔森氏菌中 Yaps 的表达和定位。根据基因组序列预测了六种不同的 Yaps，它们可能并非全部同时表达或以相同的方式定位。更好地了解每种 yaps 表达和定位的条件将为了解其潜在功能提供有价值的线索。目的 2. 分析 Yaps 在鼠疫耶尔森毒力中的作用。每个 yap 基因的突变体将在鼠疫耶尔森氏菌 CO92 菌株中构建，然后使用小鼠感染模型测试毒力。目标 3. Yaps 的功能分析。我们将通过测试表达单个 Yaps 的大肠杆菌的各种先前与自转运蛋白相关的已知功能来开始研究 Yaps 的功能。