ROLE OF YAPS IN Y. PESTIS PATHOGENESIS

雅普病在鼠疫杆菌发病机制中的作用

• 批准号: 7812704
• 负责人: VIRGINIA L MILLER
• 金额: $ 36.76万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-05-15 至 2011-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7812704
• 关键词: Acute; Address; Amino Acids; Animal Model; Antibodies; Bacterial Adhesins; Biological; Biological Assay; Bioterrorism; Blood; Cell surface; Computer Simulation; Country; Cytotoxin; Data; Disease; Disease Outbreaks; Enterobacteriaceae; Escherichia coli; Fever; Fleas; Future; Genome; Goals; Histology; Human; In Vitro; Individual; Infection; Insecta; Laboratories; Life Cycle Stages; Lung; Measures; Membrane; Modeling; Molecular; Molecular Genetics; Mus; Mutation; N-terminal; Organism; Outcome Study; Pathogenesis; Peptide Hydrolases; Peptide Signal Sequences; Phenotype; Plague; Play; Pneumonic Plague; Probability; Production; Proteins; Public Health; Reporter; Rodent; Role; Route; Septicemic plague; Sequence Analysis; Structure; Surface; System; Testing; Trees; Vaccines; Virulence; Virulent; Western Blotting; Work; Yersinia pestis; base; cytokine; genetic manipulation; human morbidity; in vivo; interest; mortality; mouse model; mutant; novel; pathogen; research study; sucking; therapeutic target; tool; transmission process; vaccine development

Yersinia pestis is a Gram-negative bacterial human pathogen that is the causative agent of plague. Historically Y. pestis has been responsible for significant human morbidity and mortality. However, Y. pestis is typically a pathogen of rodents, and is unique among the Enterobacteriaceae in using a blood-sucking insect (flea) for transmission. Y. pestis infection in humans is an acute febrile disease that can have a number of different presentations depending upon the route of inoculation. Although environmental outbreaks of plague in the developed world largely have been controlled, the possible use of this pathogen as an agent of bioterrorism, as well as the re-emergence of plague in several countries, 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. A group of proteins that have a high likelihood of contributing to Y. pestis pathogenesis are the autotransporter proteins (ATs). AT proteins consist of three basic domains: a N-terminal signal sequence, a ¿passenger domain¿ and finally a β-domain at the C-terminus which facilitates translocation of the passenger domain across the outer membrane. The passenger domain provides the functional activity and new activities are being attributed to these proteins as more of them are studied in detail. While the β-domains are relatively conserved between ATs the passenger domains can vary significantly. In silico analyses have identified ten AT proteins of Y. pestis (designated yaps/Yaps). The Yaps are of interest to study for a number of reasons. First, while many putative AT proteins have been identified by in silico analyses, relatively few have been studied experimentally and even fewer have been studied in detail. With Y. pestis we have the advantage of being able to pair sophisticated molecular genetic manipulations with an animal model that is actually a natural host for this pathogen. Second, many of the Yaps by sequence analysis are relatively divergent when evolutionary trees of ATs are constructed and thus, the likelihood of identifying new functions by studying this group of ATs is high. Third, analysis of the Yaps may help identify a useful therapeutic target or candidate for inclusion in upcoming Y. pestis vaccine cocktails. We have preliminary data indicating that all ten of the yaps are expressed not only during laboratory conditions but also during infection. We also have constructed deletion mutants in seven yaps in a fully virulent Y. pestis strain and have begun testing the effect of these mutations on virulence. Four yap mutants clearly have phenotypes in a bubonic plague model of infection; these results are consistent with our hypothesis that the yaps play a role in pathogenesis. Our long term goal is to understand at a molecular level the role(s) of the individual Yaps in disease and the Y. pestis life cycle. Thus, we propose the following specific aims: (1) Analysis of the localization and expression of Yaps in Y. pestis; (2) Analysis of the role of Yaps in virulence; (3) Analysis of potential functions of the Yaps.

耶尔森氏菌是一种革兰氏阴性细菌人类病原体，是瘟疫的灾难性药物。从历史上看，佩斯蒂斯（Y. Pestis）一直负责人类的大量发病率和死亡率。然而，Y. Pestis通常是啮齿动物的病原体，在肠杆菌科中使用血液吸血绝缘（跳蚤）进行透射是独一无二的。 Y. pestis infection in humans is an acute febrile disease that can have a number of different presentations depending upon the route Although environmental outbreaks of plague in the developed world largely have been controlled, the possible use of this pathogen as an agent of bioterrorism, as well as the re-emergence of plague in several countries, necessary to provide a more detailed understanding of the pathogenesis of Y. pestis to facilitate the identification of targets for疫苗开发和治疗。目前没有疫苗。一组蛋白质很有可能有助于Y. Pestis 发病机理是自转运蛋白（ATS）。在蛋白质中，由三个基本域组成：N末端信号序列，A乘客结构域和C-terminus处的β-域，该域的设施在整个外膜上的设施转移。乘客域提供了功能活动，并且由于其中更多的详细研究，因此将新活动归因于这些蛋白质。虽然β-域在ATS之间相对保守，但乘客域可能会有很大的变化。在计算机分析中，已经在Y. Pestis的蛋白质（指定的YAPS/YAPS）鉴定了十个。出于多种原因，yaps很感兴趣。首先，尽管在硅分析中已经确定了许多假定的蛋白质，但 对实验进行了相对较少的研究，详细研究了更少的研究。借助Y. Pestis，我们的优点是能够将复杂的分子遗传操作与动物模型配对，这实际上是该病原体的天然宿主。其次，当构建ATS的进化树时，许多按序列分析的YAP相对不同，因此，通过研究这组ATS来识别新功能的可能性很高。第三，对YAP的分析可能有助于确定有用的治疗靶标或候选者，以纳入即将到来的Y. Pestis疫苗鸡尾酒。我们有初步数据，表明所有十个YAP不仅在实验室条件下而且在感染过程中表达。我们还以完全毒性的y菌株构建了七个yap的缺失突变体，并开始测试效果 这些关于病毒的突变。四个YAP突变体在感染的气泡鼠疫模型中显然具有表型。这些结果与我们的假设是一致的，即YAP在发病机理中起作用。我们的长期目标是在分子水平上了解各个YAP在疾病和PESTIS生命周期中的作用。这是我们提出以下特定目的的：（1）分析Y. Pestis中YAP的定位和表达； （2）分析YAP在病毒中的作用； （3）分析YAP的潜在功能。