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

项目摘要/摘要 非霍乱弧菌(NCV)对人类健康和食品安全构成显著且日益严重的威胁。 弧菌所形成的生物膜具有很强的生命力，与宿主感染高度相关。生物膜被紧密地 基质相关细菌的调节群落，是细菌致病的主要组成部分 包括抗药性。由于估计75%的细菌感染涉及生物被膜，因此更好地 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。这种蛋白质在NCVS中是保守的 并控制共生多糖(SYP)的生产，SYP是生物膜基质的一种成分，参与 宿主感染。根据初步发现，与Well相比，这种蛋白质具有非典型的作用模式 表征了同系物。我的基本假设是这种NCV生物膜调节蛋白使用了一种新的 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)利用生物化学来探测蛋白质的功能 和结构特征研究，以及2)通过确定其在监管网络中的位置 有约束力的伙伴。这项拟议的研究具有创新性，因为它关注的是一种对生物膜至关重要的系统 到目前为止，这个系统只用细胞和遗传学方法进行了研究。这个 这里提出的结构-功能方法将提供填补我们知识空白所需的基本信息。 该项目意义重大，因为它将提供对宿主相关的监管的更深层次的理解 NCV感染的威胁不断上升，导致生物被膜。最终，提出的工作将回答有关的关键问题 对调节寄主相关的关键和保守途径中的蛋白质的分子机制 生物膜。K22奖项的支持将促进我过渡到独立调查员，因为它为我提供了 有资源来产生数据和出版物，这将增强我对未来资金的竞争力。

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.