Diversity and evolution of P syringae type III effectors

丁香假单胞菌III型效应子的多样性和进化

基本信息

批准号：
7158585
负责人：
JEFFERY L. DANGL
金额：
$ 27.62万
依托单位：
UNIV OF NORTH CAROLINA CHAPEL HILL
依托单位国家：
美国
项目类别：
财政年份：
2004
资助国家：
美国
起止时间：
2004-01-01 至 2008-05-04
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7158585
关键词：
Affect Agriculture Alleles Animal Model Animals Arabidopsis Arabidopsis Proteins Bacteria Biochemical Bioinformatics Biological Assay C-terminal Cell Separation Cell physiology Cells Cellular biology DNA Microarray Chip DNA Sequence Data Disease Disease Resistance Dissection Ensure Evolution Facility Construction Funding Category Flow Cytometry Gene Family Genes Genetic Genetic Screening Genetic Transcription Genome Genomics Green Fluorescent Proteins Health Host Defense Host resistance Human Immune response Injection of therapeutic agent Libraries Metabolism Methodology Microbiology Modeling Molecular Evolution Monitor N-terminal Open Reading Frames Operon Pathogenesis Pathogenicity Phenotype Phylogenetic Analysis Plants Proteins Pseudomonas syringae Role Salmonella Sequence Analysis Shigella Signal Transduction Southern Blotting System Testing Transgenic Organisms Type III Secretion System Pathway Virulence Virulence Factors Virulent Work Yersinia base biodefense defense response enteropathogenic Escherichia coli fluorescence activated cell sorter device pathogen pathogenic Escherichia coli pathogenic bacteria promoter response tool

项目摘要

DESCRIPTION (provided by applicant): Identification of virulence factors is a necessary first step to study the mechanisms and evolution of pathogenicity. We propose to compare the type III disease effector proteins from a phylogenetically diverse set of plant pathogenic Pseudomonads chosen to represent the most broad distribution of the species possible. P. syringae is an ideal model organism to study the distribution and evolution of type III effectors. Type III effectors interact with cellular host targets, and modulate host defense responses or metabolism in a manner conducive to pathogen proliferation. Most of the P. syringae effectors have not yet been characterized, and we remain naive as to the collective diversity of host cellular functions they manipulate. Although the functions of P. syringae effector proteins during disease remain poorly understood, they can often be monitored via their phenotypes following in planta expression of a given type III effector. Bacterial pathogens of both animals, such as Salmonella spp., Yersinia spp., Shigella spp. and pathogenic E. coli also rely on type III secretion systems for pathogenesis. Thus, our results will inform studies of these bacteria and their animal hosts, including humans. Because P. syringae is pathogenic on a variety of distantly related plant hosts, many of which can be genetically manipulated, our system has advantages over models of type III pathogenesis of animals, which generally focus only on strains pathogenic on phylogenetically related mammalian hosts. We have devised and implemented a high throughput Fluorescence Activated Cell Sorter (FACS) based experimental approach to capture all of the type III effectors from the genome of any given P. syringae isolate. We chose 13 P syringae isolates that are pathogens of an evolutionarily diverse set of host plants. We intend to sieve through these 13 genomes to describe their suites of type III effectors. We further intend to begin dissection of their effects on host cell biology, using the easily manipulated Arabidopsis plant as a model where appropriate. This proposal is highly interdisciplinary, drawing on methodologies and expertise in microbiology, bioinformatics and plant-pathogen interactions in conjunction with high throughput bacterial cell sorting using the FACS. Our work will broadly impact the understanding of a widely distributed pathogenicity mechanism that affects both human health and agriculture. Additionally, our work may have relevance in biodefense with respect to basic understanding of pathogenesis in bacteria that can potentially be weaponized.

描述（由申请人提供）：识别毒力因子是研究致病性机制和演变的必要第一步。我们建议比较从系统发育多样的植物致病性假单胞菌中选择的III型疾病效应蛋白，以代表可能的物种最广泛的分布。丁香假单胞菌是研究III型效应子的分布和演变的理想模型生物。 III型效应子与细胞宿主靶标相互作用，并以有利于病原体增殖的方式调节宿主防御反应或代谢。尚未表征大多数丁香假单胞菌效应子，并且对于它们操纵的宿主细胞功能的集体多样性，我们仍然很幼稚。尽管在疾病期间，在疾病期间的春季假单胞菌效应子蛋白的功能仍然很少了解，但通常可以通过其表型来监测它们在给定III型效应子的植物表达中。两种动物的细菌病原体，例如沙门氏菌属，耶尔森氏菌，志贺氏菌。病原大肠杆菌还依赖于III型分泌系统进行发病机理。因此，我们的结果将为包括人类在内的这些细菌及其动物宿主的研究提供信息。由于丁香假单胞菌对各种远距离相关的植物宿主具有致病性，其中许多宿主可以通过遗传操纵，因此我们的系统比动物的III型发病机理的模型具有优势，这些模型通常仅着重于对系统上相关的哺乳动物宿主的致病性菌株。我们已经设计并实施了基于高吞吐量的荧光活化细胞分落蛋白（FACS）的实验方法，以从任何给定的丁香假单胞菌分离株的基因组中捕获所有III型效应子。我们选择了13种P丁香分离株，这些分离株是一组进化上多样的宿主植物的病原体。我们打算筛选这13个基因组，以描述其III型效应子的套件。我们进一步打算使用易于操纵的拟南芥植物作为适当的模型开始解剖其对宿主细胞生物学的影响。该建议是高度跨学科的，借鉴了微生物学，生物信息学和植物病原体相互作用的方法和专业知识，并结合使用FACS的高吞吐量细菌细胞分选。我们的工作将广泛影响对影响人类健康和农业的广泛分布的致病机制的理解。此外，我们的工作可能与生物陷入困境有关，就可能被武器化的细菌中的发病机理的基本理解而言。