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

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

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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，称为细菌或噬菌体，是最丰富的生物学实体之一行星。这些病毒在基因组含量，感染机制，复制周期和结构。随着对使用细菌量对抗抗生素耐药细菌的兴趣越来越多，它是了解这些病毒如何感染宿主的机制所必需的；复制并调节他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/或存储条件。但是，我们当前的模型系统固有地仅代表噬菌体的一部分多样性。这项工作的长期目标是开发一个额外的细菌系统 - 摩格病毒 - 这在环境和临床上有用的品质似乎无处不在。但是，他们才最近我们之所以被孤立，是因为它们通常不感染细菌大肠杆菌的模型种类和沙门氏菌肠。取而代或机会主义的病原体，例如柠檬酸菌。因为这些病毒是规则裂解和特异性的对于腹泻最常见的病因学剂之一，S。flexneri，它们可用于控制这一点食物或水中的细菌种类，或用于治疗抗生素耐药性感染。但是，目前我们缺乏充分了解其生物过程。 Mooglevirus具有不同的特征与其他特征噬菌体，包括半特异性宿主范围，其基因组中编码的大量TRNA和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案与更彻底的模型系统相比，感染和持续存在的策略。目标的目标因此，该具体建议是为了确定志氏菌的机制用于识别穆格利病毒他们的宿主在翻译层面调节病毒和宿主基因的表达，然后组装成新的可以在环境中持续存在的粒子。该提案的预期结果是：1）噬菌体受体结合蛋白与原发性和次要的临界相互作用区域 S. flexneri宿主的受体，并确定附着和进入的动力学； 2）完整的肋骨 - 来自各种环境和感染条件的SEQ和RNA-SEQ数据集，导致机械了解噬菌体感染如何改变噬菌体和宿主基因的转化效率； 3）广泛 Mooglevirus Capsid的已解决的组装途径，具有蛋白质或蛋白质结构域的指示负责影响衣壳稳定性。这项工作将对基本生物学产生影响，扩大我们的已知的噬菌体策略以及感染和持久性的机制的曲目；和医疗应用程序，告知这些噬菌体本身如何应用于S. flexneri感染诊所，或者如何使用其特性来设计新颖的噬菌体，以用于医疗或工业应用。