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

基本信息

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
项目类别：
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

项目摘要

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。这些病毒在基因组内容、感染机制、复制周期和功能方面差异很大。随着人们对使用噬菌体对抗抗生素耐药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；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。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—— 然而，它们在环境和临床有用的品质中似乎无处不在。被分离出来是因为它们通常不感染模型细菌大肠杆菌和相反，这些病毒通常针对致病菌福氏志贺氏菌进行分离。或机会性病原体，例如弗氏柠檬酸杆菌，因为这些病毒具有专性裂解性和特异性。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然而，目前我们缺乏食物或水中的细菌种类，或用于治疗抗生素耐药性感染的药物。对其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，以及这项工作的中心假设是这些病毒使用替代的衣壳和基因组大小。与更彻底表征的模型系统相比，感染和持续的策略。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，并组装成新的该提案的预期结果是： 1) 识别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。已知的噬菌体策略以及感染和持久性机制的全部内容；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。