Regulatory mechanism of novel host-relevant biofilm formation protein in non-Cholera Vibrio species

项目总结/摘要 非霍乱弧菌（NCV）物种对人类健康和食品安全构成显著且日益严重的威胁。 由弧菌属物种形成的生物膜是稳健的并且与宿主感染高度相关。生物膜紧密地 基质相关细菌的调节群落，是细菌致病的主要成分 包括抗药性。由于估计有75%的细菌感染涉及生物膜，因此必须更好地 了解生物膜是如何形成的在这个应用中，我建议阐明一个分子机制， 控制NCV生物膜形成所必需的不寻常的调节蛋白。该蛋白在NCV中是保守的 并控制共生多糖（Syp）的产生，Syp是生物膜基质的一种成分， 宿主感染基于初步的发现，这种蛋白质具有一种非典型的作用模式， 特征同系物。我的基本假设是，这种NCV生物膜调节蛋白使用一种新的 控制生物膜形成的机制，并将挑战我们目前对这一蛋白质家族的理解。 我的假设将通过两个具体的目标来解决：1）使用生物化学方法探测蛋白质的功能， 和结构表征研究，以及2）通过识别其在监管网络中的位置 有约束力的伙伴这项拟议中的研究是创新的，因为它关注的是一个对生物膜至关重要的系统。 形成，迄今为止，该系统仅使用细胞和遗传方法进行了研究。的 本文提出的结构-功能方法将为填补我们的知识空白提供必要的信息。 该项目是重要的，因为它将提供一个更深层次的了解，调节主机相关的 NCV感染的威胁日益增加。最终，拟议的工作将回答有关的关键问题， 在一个关键的和保守的途径，调节宿主相关的蛋白质的分子机制， 生物膜K22奖的支持将帮助我过渡到独立调查员， 有资源产生数据和出版物，这将加强我的竞争力，为未来的资金。

PROJECT SUMMARY/ABSTRACT Non-cholera Vibrio (NCV) species represent a notable and increasing threat to human health and food safety. The biofilms formed by Vibrio species are robust and highly relevant to host infection. Biofilms are tightly regulated communities of matrix-associated bacteria and are a major component of bacterial pathogenesis including drug resistance. Since an estimated 75% of bacterial infections involve biofilms, it is crucial to better understand how biofilms are formed. In this application, I propose to elucidate the molecular mechanism of an unusual regulatory protein necessary for controlling NCV biofilm formation. This protein is conserved in NCVs and controls the production of symbiosis polysaccharide (Syp), a component of the biofilm matrix involved in host infection. Based on preliminary findings, this protein has an atypical mode of action compared to well characterized homologs. My fundamental hypothesis is that this NCV biofilm regulator protein uses a novel mechanism to control biofilm formation and will challenge our current understanding of this family of proteins. My hypothesis will be addressed through two specific aims: 1) probing the protein’s function using biochemical and structural characterization studies, and 2) uncovering its position in the regulatory network by identifying binding partners. The proposed research is innovative because it focuses on a system that is crucial for biofilm formation, and to date, the system has only been investigated using cellular and genetic approaches. The structure-function approach proposed here will provide essential information needed to fill our knowledge gaps. The project is significant because it will provide a deeper understanding of the regulation of host-associated biofilms from the rising threat of NCV infection. Ultimately, the work proposed will answer key questions related to the molecular mechanisms of a protein within a critical and conserved pathway that regulates host-relevant biofilms. Support from this K22 award will facilitate my transition to an independent investigator by providing me with resources to generate data and publications that will strengthen my competitiveness for future funding.