Autotransporter proteins and virulence of Y. pestis

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，由于高死亡率和严重的疾病引起的Y。pestis，Y.鼠疫已成为生物战和生物恐怖主义的潜在病原体。虽然鼠疫的环境爆发在很大程度上已通过使用现代公共卫生措施得到控制，但这种病原体作为战争和恐怖媒介的潜在用途需要更详细地了解Y.鼠疫，以促进确定疫苗开发和治疗的目标。目前还没有针对淋巴腺鼠疫或肺鼠疫的疫苗。对最近发表的Y. pestis表明Y.鼠疫菌可能表达六种不同的自转运蛋白（AT）（命名为耶尔森氏菌自转运蛋白的雅普）。AT蛋白已经在多种病原体中被鉴定，并且正如其名称所暗示的，蛋白质的一级序列足以指导穿过细菌的外膜的运输。此外，在已知AT功能的情况下，这些蛋白质一致地与毒力相关。在Y.鼠疫菌基因组的研究对于理解鼠疫菌与鼠疫菌的相互作用具有重要意义。鼠疫及其宿主以及潜在的疫苗候选者。因此，开始研究Y.我们提出以下具体目标：目标1。Yaps基因在Y.鼠疫从基因组序列预测六种不同的Yap，并且它们可能不都同时表达或以相同的方式定位。更好地理解每一个yap表达和定位的条件将为其潜在功能提供有价值的线索。目标2.分析Yaps在Y.t.毒力中的作用。鼠疫将在Y的CO92菌株中构建每个雅普基因的突变体。鼠疫杆菌，然后使用小鼠感染模型测试毒力。目标3。雅浦人的功能分析。我们将开始通过测试E.大肠杆菌表达单个Yaps的各种已知功能，以前与自转运蛋白。

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.