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.

项目摘要/摘要 在自然栖息地中，细菌受到被感染的病毒，称为噬菌体的轰击。感染宿主后，噬菌体 必须承担两种生活方式之一：裂解性的，噬菌体保留在宿主中并传给 噬菌体复制的后代或裂解，杀死宿主并扩散到新细胞。噬噬菌体已经被想到 仅仅是针对宿主应力和DNA损伤的响应，从裂解性过渡到裂解。来自的新研究 巴斯勒实验室透露，噬菌体可以监视宿主通信分子，称为 自动诱导仪。在称为Quorum传感器的过程中，细菌产生，释放和检测自动诱导剂，并在 响应，编排小组行为。检测到宿主产生的自动诱导剂的群体感应响应噬菌体 并利用他们获得的信息来推动其裂解赖以散失的生活方式过渡。这些最近的发现 定位我，发现噬菌体如何操纵细菌宿主以及对宿主的后果，多 宿主是成员的物种细菌群落，以及所有实体的真核生物宿主 居住。我的研究的总体目标是定义颤音之间的跨域通信 VP882，第一个发现在法定人数上“窃听”的噬菌体及其宿主，全球病原体颤音 霍乱，推出了噬菌体裂解周期。结合遗传，生化和结构 方法，我将确定这种宿主阶段化学通信的基础的分子机制 过程。首先，我将从巴斯勒实验室的专家那里学习细菌遗传学的技能，并进行遗传 屏幕以识别法定感应诱导的噬菌体裂解周期的复制品。第二，我将使用生化 定量表征Quorum-两个关键信号分量之间相互作用的方法 感应引起的噬菌体裂解途径。最后，我将依靠我的结构生物学背景来解决 这些相同信号分量的结构单独和复杂，使原子级分辨率启用 了解噬菌体经历生活方式过渡所需的相互作用。理想的结果 我的研究是对域间化学交流的机械理解和新的可能性 噬菌体疗法的发展。磨练我在细菌遗传学，蛋白质生物化学和 在我的博士后培训过程中，大分子晶体学将使我能够发起 顶级研究机构的独立研究计划。