Discovery of novel signaling components targeted by Vibrio

描述（由申请人提供）：本提案的总体目标是鉴定在胃肠道病原体副溶血性弧菌感染肠道期间靶向的新型宿主信号成分。副溶血性弧菌是在太平洋和大西洋发现的与食用受污染海产品有关的肠胃炎暴发的主要病原体。了解病原体如何破坏正常的体内平衡并削弱宿主的防御反应对于我们理解胃肠道细胞利用的信号通路至关重要。最近对副溶血性v基因组的测序显示，存在一个编码多种毒素的致病性岛（PAII）、一个III型分泌系统（TTSS）、一个可识别的效应物VopA，以及一些被认为编码新型效应物的其他未表征的开放阅读框。先前对效应物的研究表明，这些蛋白通过捕获或模仿对宿主细胞调节至关重要的真核调节功能而进化。这些效应物利用的机制的发现为真核细胞信号传导的基本机制提供了见解。我们最初的研究集中在属于yopj样毒力因子家族的效应物VopA的表征上。我们正在使用VopA作为副溶血性弧菌的代表性效应器来设计一个系统，用于分析编码在PAII中的未知效应器。通过鉴定副溶血性弧菌表达的新型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关键成分。这笔拨款将用于资助涉及开发体外副溶血性弧菌感染系统的研究，以研究由一种革兰氏阴性海洋细菌产生的致病效应物，这种细菌在世界范围内导致食物中毒。此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.