Quorum-sensing mediated communication between pandemic Vibrio cholerae and phage VP882

群体感应介导大流行霍乱弧菌和噬菌体 VP882 之间的通讯

• 批准号: 10601559
• 负责人: Grace Beggs
• 金额: $ 6.91万
• 依托单位: PRINCETON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-02-01 至 2025-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10601559
• 关键词: Bacteria; Bacteriophages; Behavior; Binding; Biochemical; Cells; Chemicals; Chromosomes; Communication; Communities; Complex; Crystallization; Crystallography; Cues; Cytolysis; DNA Damage; Detection; Development; Engineering; Foundations; Genes; Genetic; Genetic Screening; Genetic study; Goals; Habitats; Individual; Industry; Infection; Institution; Intercept; Investigation; Label; Laboratories; Learning Skill; Length; Life Style; Light; Lysogeny; Lytic; Lytic Phase; Maintenance; Mass Spectrum Analysis; Mediating; Methods; Molecular; Monitor; Mutagenesis; Outcome; Pathway interactions; Phage Receptors; Physiological; Positioning Attribute; Process; Production; Protein Biochemistry; Proteins; Recombinants; Reporter; Repression; Repressor Proteins; Research; Resolution; Role; Sensory; Signal Transduction; Stress; Structure; Vibrio cholerae; Vibrio cholerae infection; Virus; Work; arms race; bacterial community; bacterial genetics; combat; follow-up; human disease; imaging study; in vivo; insight; member; offspring; overexpression; pandemic disease; pathogen; post-doctoral training; programs; promoter; quorum sensing; response; skills; structural biology; therapy development

PROJECT SUMMARY/ABSTRACT Bacteria are bombarded by infecting viruses, called phages, in natural habitats. Upon infection of a host, phages must undertake one of two lifestyles: lysogeny where the phage remains in the host and is passed down to offspring, or lysis where the phage replicates, kills the host, and spreads to new cells. Phages have been thought to transition from lysogeny to lysis exclusively in response to host stress and DNA damage. New research from the Bassler laboratory has revealed that phages can monitor host communication molecules, called autoinducers. In a process called quorum sensing, bacteria produce, release, and detect autoinducers, and in response, orchestrate group behaviors. Quorum-sensing-responsive phages detect host-produced autoinducers and exploit the information they garner to drive their lysis-lysogeny lifestyle transitions. These recent findings position me to discover how phages manipulate bacterial hosts and the consequences to the host, to the multi- species bacterial community of which the host is a member, and to the eukaryotic host in which all the entities reside. The overarching goal of my research is to define how cross-domain communication between vibriophage VP882, the first phage discovered to “eavesdrop” on quorum sensing, and its host, the global pathogen Vibrio cholerae, launches the phage lytic cycle. Using a combination of genetic, biochemical, and structural approaches, I will identify the molecular mechanisms underlying this host-phage chemical communication process. First, I will learn skills in bacterial genetics from experts in the Bassler laboratory and conduct a genetic screen to identify the repressor of the quorum-sensing-induced phage lytic cycle. Second, I will use biochemical methods to quantitatively characterize interactions between two key signaling components in the quorum- sensing-induced phage lysis pathway. Lastly, I will rely on my background in structural biology to solve the structures of these same signaling components, individually and in complex, enabling atomic-level-resolution understanding of the interactions required for the phage to undergo lifestyle transitions. The ideal outcomes of my research are a mechanistic understanding of inter-domain chemical communication and new possibilities for development of phage therapies. Honing my skills in bacterial genetics, protein biochemistry, and macromolecular crystallography over the course of my postdoctoral training will enable me to launch an independent research program at a top-tier research institution.

项目总结/摘要 在自然环境中，细菌受到感染性病毒的轰击。在感染宿主后， 必须采取两种生活方式之一：溶原性，其中噬菌体留在宿主体内，并传递给 噬菌体复制、杀死宿主并传播到新细胞的裂解。噬菌体一直被认为 从溶原性到溶解性的转变仅仅是对宿主应激和DNA损伤的反应。的新研究 Bassler实验室已经发现，Escherichia coli可以监控宿主的通讯分子， 自体诱导物。在一个称为群体感应的过程中，细菌产生，释放和检测自动诱导物，并在 回应，协调群体行为。群体感应反应性DNA检测宿主产生的自诱导物 并利用他们收集的信息来驱动他们的溶细胞-溶原性生活方式的转变。这些最新的发现 我的位置，以发现如何操纵细菌宿主和后果的主机，以多- 物种细菌群落，其中宿主是一个成员，并真核宿主，其中所有实体 居住。我研究的首要目标是确定弧菌噬菌体之间的跨域通信 VP 882，发现的第一个“窃听”群体感应的噬菌体，及其宿主，全球病原体弧菌 霍乱毒素启动噬菌体裂解周期。综合运用遗传、生化和结构 方法，我将确定这种宿主-噬菌体化学通讯的分子机制 过程首先，我将向巴斯勒实验室的专家学习细菌遗传学方面的技能，并进行遗传学研究。 筛选以鉴定群体感应诱导的噬菌体裂解周期的阻遏物。第二，我会用生化 定量表征群体中两个关键信号组分之间相互作用的方法， 感测诱导的噬菌体裂解途径。最后，我将依靠我在结构生物学方面的背景来解决这个问题。 这些相同的信号成分的结构，单独和复杂，使原子级分辨率 了解噬菌体经历生活方式转变所需的相互作用。理想的结果 我的研究是对域间化学通讯的机械理解， 噬菌体疗法的发展。磨练我在细菌遗传学，蛋白质生物化学， 在我的博士后培训过程中，大分子晶体学将使我能够开展一项 在顶级研究机构的独立研究项目。