Discovery of novel signaling components targeted by Vibrio

描述（由申请人提供）： 这项建议的总体目标是确定新的主机信号组件，在肠道感染的胃肠道病原体副溶血性弧菌的目标。副溶血性弧菌是导致胃肠炎爆发的主要病原体，与太平洋和大西洋发现的受污染海鲜消费有关。了解病原体是如何破坏正常的体内平衡和削弱宿主的防御反应是至关重要的，我们了解的信号转导途径所利用的胃肠道细胞。最近对副溶血性弧菌基因组的测序揭示了编码许多毒素的致病性岛（PAII）、III型分泌系统（TTSS）、一种可识别的效应物VopA和许多其他被认为编码新效应物的未表征的开放阅读框架的存在。先前对效应子的研究表明，这些蛋白质是通过捕获或模仿真核生物的调节功能而进化的，这些调节功能对宿主细胞的调节至关重要。这些效应物中的每一个所利用的机制的发现提供了对真核细胞信号传导的基本机制的见解。我们的初步研究集中在属于YopJ样毒力因子家族的效应子VopA的表征上。我们使用VopA作为副溶血性弧菌的代表性效应子，设计了一个分析PAII内编码的未知效应子的系统。通过鉴定副溶血性弧菌表达的新毒力因子并揭示其在真核细胞中的靶点，我们将发现宿主信号通路的新的关键组分。这笔赠款将用于资助参与开发体外副溶血性弧菌感染系统的研究，以研究导致全球食物中毒的革兰氏阴性海洋细菌产生的致病效应物。本次申请的结果将用作提交未来RO1申请的初步数据。

DESCRIPTION(provided by applicant): The overall goal of this proposal is to identify novel host signaling components that are targeted during infection of the gut by the gastrointestinal pathogen Vibrio parahaemolyticus. V. parahaemolyticus is a major agent responsible for gastroenteritis outbreaks associated with the consumption of contaminated seafood found in both the Pacific and Atlantic oceans. Understanding how pathogens disrupt normal homeostasis and cripple the host defense response is critical for our understanding of signaling pathways utilized by gastrointestinal cells. Recent sequencing of the V. parahaemolyticus genome has revealed the presence of a pathogenicity island (PAII) that encodes a number of toxins, a type III secretion system (TTSS), one recognizable effector, VopA, and a number of other uncharacterized open reading frames thought to encode novel effectors. Previous studies on effectors demonstrate that these proteins have evolved by capturing or mimicking eukaryotic regulatory functions that are critical for host cell regulation. The discovery of the mechanism utilized by each of these effectors has provided insights into the essential mechanisms of eukaryotic cellular signaling. Our initial studies have focused on the characterization of the effector VopA that belongs to the YopJ-like family of virulence factors. We are using VopA as a representative effector from V. parahaemolyticus to design a system for analysis of unknown effectors encoded within PAII. By identifying the novel virulence factors expressed by V. parahaemolyticus and revealing their targets in a eukaryotic cell, we will uncover new critical components of host signaling pathways. This grant will be used to fund research involved in developing an in vitro V. parahaemolyticus infection system to study pathogenic effectors produced by a gram-negative marine bacterium that is responsible for causing food poisoning worldwide. Results from this application will be used as preliminary data for the submission of a future RO1 application.