Mechanisms of interaction between bacteriophage and their hosts throughout the infection cycle

整个感染周期中噬菌体与其宿主之间的相互作用机制

• 批准号: 10644004
• 负责人: Sarah M Doore
• 金额: $ 35.39万
• 依托单位: UNIVERSITY OF FLORIDA
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-06-13 至 2027-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10644004
• 关键词: Address; Affect; Affinity; Antibiotic Resistance; Bacteria; Bacterial Infections; Bacteriophages; Binding; Binding Proteins; Biochemical; Biological; Biological Assay; Biological Models; Biological Process; Biology; Capsid; Cells; Cessation of life; Characteristics; Citrobacter freundii; Clinic; Clinical; Clustered Regularly Interspaced Short Palindromic Repeats; Collaborations; Combating Antibiotic Resistant Bacteria; Communicable Diseases; Complication; Cryo-electron tomography; Cryoelectron Microscopy; Data; Data Analyses; Data Set; Developing Countries; Diarrhea; Disease; Dysentery; Engineering; Enteral; Environment; Escherichia coli; Etiology; Evolution; Florida; Food; Functional disorder; Gastrointestinal tract structure; Gene Expression; Gene Expression Regulation; Genes; Genetic; Genetic Transcription; Genome; Geometry; Goals; Gram-Negative Bacteria; Health; Human; Human body; In Vitro; Industrialization; Infection; Kinetics; Knock-out; Lipopolysaccharides; Location; Lytic; Mass Spectrum Analysis; Measures; Medical; Membrane Proteins; Michigan; Mission; Modeling; Multicenter Studies; Mutagenesis; National Institute of Allergy and Infectious Disease; Organism; Outcome; Pathway interactions; Phage Receptors; Planets; Preventive therapy; Preventive treatment; Production; Property; Protein Analysis; Proteins; RNA Sequences; Research; Resolution; Risk; Salmonella enterica; Series; Serotyping; Shigella; Shigella Infections; Shigella flexneri; Specificity; Stress; Structure; Surface; System; Tail; Techniques; Tertiary Protein Structure; Testing; Time; Transfer RNA; Universities; Viral; Virus; Water; Work; World Health Organization; antibiotic resistant infections; combat; crosslink; experimental study; fitness; gel electrophoresis; genetic analysis; gut bacteria; imaging modality; in vivo; innovation; interest; knock-down; mortality; mutant; next generation sequencing; novel; novel therapeutics; opportunistic pathogen; particle; pathogen; pathogenic bacteria; receptor; recruit; ribosome profiling; transcriptome sequencing

Bacterial viruses, known as bacteriophages or phages, are among the most abundant biological entities on the planet. These viruses vary broadly in terms of genome content, infection mechanisms, replication cycles, and structure. With an increasing interest in using bacteriophages to combat antibiotic resistant bacteria, it is necessary to understand mechanisms of how these viruses infect their hosts; replicate and regulate both their genes and their hosts’ genes; and how they persist in various environments, including the human body and/or storage conditions. However, our current model systems inherently only represent a fraction of bacteriophage diversity. The long-term goal of this work is to develop an additional bacteriophage system—the Moogleviruses— that appear to be ubiquitous in the environment and clinically useful qualities. They have, however, only recently been isolated because they do not commonly infect the model species of bacteria Escherichia coli and Salmonella enterica. Instead, these viruses are often isolated against the pathogenic bacteria Shigella flexneri or opportunistic pathogens such as Citrobacter freundii. Because these viruses are obligately lytic and specific to one of the most common etiological agents of diarrhea, S. flexneri, they could be used for controlling this species of bacteria in food or water, or for treating antibiotic-resistant infections. At this time, however, we lack sufficient understanding of their biological processes. Moogleviruses have distinct characteristics versus other bacteriophages, including a semi-specific host range, a large number of tRNAs encoded in their genomes, and uncommon capsid and genome sizes. The central hypothesis of this work is that these viruses use alternative strategies to infect and persist compared to more thoroughly characterized model systems. The objectives of this specific proposal are therefore to determine the mechanisms Shigella-infecting Moogleviruses use to identify their hosts, modulate the expression of viral and host genes on a translational level, and assemble into new particles that can persist in the environment. The expected outcomes of this proposal are: 1) the identification of critical interacting regions between the phage receptor-binding proteins and both primary and secondary receptors on the S. flexneri host, along with determining the kinetics of attachment and entry; 2) complete Ribo- seq and RNA-seq datasets from a variety of environments and infection conditions, leading to a mechanistic understanding of how phage infection alters translational efficiency of phage and host genes; and 3) a broadly resolved assembly pathway of the Mooglevirus capsid, with an indication of proteins or protein domains responsible for affecting capsid stability. This work will have impacts for both basic biology, expanding our repertoire of known bacteriophage strategies and mechanisms for infection and persistence; and medical application, informing how these phages themselves could be applied to combat S. flexneri infections in the clinic, or how their properties could be used to engineer novel phages for medical or industrial applications.

细菌病毒，称为噬菌体或噬菌体，是地球上最丰富的生物实体之一。 行星。这些病毒在基因组内容、感染机制、复制周期和 结构。随着人们对使用噬菌体对抗抗生素耐药性细菌的兴趣日益浓厚， 有必要了解这些病毒如何感染宿主的机制；复制并调节它们的 基因及其宿主的基因；以及它们如何在各种环境中持续存在，包括人体和/或 储存条件。然而，我们当前的模型系统本质上仅代表噬菌体的一小部分 多样性。这项工作的长期目标是开发一种额外的噬菌体系统——莫格尔病毒—— 这似乎在环境和临床上有用的品质中无处不在。然而，他们直到最近才 被分离出来是因为它们通常不感染模型细菌大肠杆菌和 肠道沙门氏菌。相反，这些病毒通常是针对致病菌福氏志贺氏菌进行分离的。 或机会致病菌，例如弗氏柠檬酸杆菌。因为这些病毒具有绝对裂解性和特异性 对于腹泻最常见的病原体之一福氏链球菌，它们可用于控制这种情况 食物或水中的细菌种类，或用于治疗抗生素耐药性感染。然而此时我们却缺乏 对它们的生物过程有足够的了解。莫格尔病毒与其他病毒相比具有独特的特征 噬菌体，包括半特异性宿主范围、其基因组中编码的大量 tRNA，以及 不常见的衣壳和基因组大小。这项工作的中心假设是这些病毒使用替代方法 与更彻底表征的模型系统相比，感染和持续的策略。的目标 因此，该具体建议是确定感染志贺氏菌的莫格尔病毒用于识别的机制 它们的宿主，在翻译水平上调节病毒和宿主基因的表达，并组装成新的 可以持续存在于环境中的颗粒。该提案的预期成果是： 1) 确定 噬菌体受体结合蛋白与初级和次级之间的关键相互作用区域 福氏链球菌宿主上的受体，以及确定附着和进入的动力学； 2) 完整的Ribo- 来自各种环境和感染条件的 seq 和 RNA-seq 数据集，从而产生了一种机制 了解噬菌体感染如何改变噬菌体和宿主基因的翻译效率； 3）广泛的 解析了莫格尔病毒衣壳的组装途径，并标明了蛋白质或蛋白质结构域 负责影响衣壳稳定性。这项工作将对基础生物学产生影响，扩大我们的研究范围 已知噬菌体策略以及感染和持久性机制的全部内容；和医疗 应用，告知如何应用这些噬菌体本身来对抗福氏链球菌感染 临床，或者如何利用它们的特性来设计用于医疗或工业应用的新型噬菌体。