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.

项目概要/摘要 在自然栖息地中，细菌会受到称为噬菌体的感染病毒的轰炸。 必须采取两种生活方式之一： 溶原性，其中噬菌体保留在宿主体内并传递给 人们认为噬菌体会在后代或裂解中复制、杀死宿主并扩散到新细胞。 专门针对宿主应激和 DNA 损伤从溶源转变为裂解。 巴斯勒实验室发现噬菌体可以监测宿主通讯分子，称为 在称为群体感应的过程中，细菌产生、释放和检测自诱导剂。 群体感应响应噬菌体检测宿主产生的自诱导剂。 并利用他们获得的信息来推动他们的裂解-溶原生活方式转变。 让我能够发现噬菌体如何操纵细菌宿主以及对宿主、对多种微生物的影响 宿主是其成员的物种细菌群落，以及所有实体都存在于其中的真核宿主 我研究的首要目标是定义噬菌体之间的跨域通信。 VP882，第一个被发现“窃听”群体感应的噬菌体，及其宿主，全球病原体弧菌 霍乱，结合遗传、生化和结构启动噬菌体裂解循环。 方法，我将确定这种宿主噬菌体化学通讯背后的分子机制 首先，我会向巴斯勒实验室的专家学习细菌遗传学的技能，并进行遗传学研究。 筛选以确定群体感应诱导的噬菌体裂解循环的阻遏物。 其次，我将使用生化。 定量表征群体中两个关键信号成分之间相互作用的方法 最后，我将依靠我的结构生物学背景来解决这个问题。 这些相同信号成分的结构，无论是单独的还是复杂的，都可以实现原子级分辨率 了解噬菌体经历生活方式转变所需的相互作用。 我的研究是对域间化学通讯的机械理解以及新的可能性 磨练我在细菌遗传学、蛋白质生物化学和噬菌体疗法方面的技能。 在我的博士后培训过程中，大分子晶体学将使我能够启动一个 顶级研究机构的独立研究项目。