OXIDATIVE STRESS RESPONSES IN PATHOGENIC PSEUDOMONAS SPECIES

致病性假单胞菌种的氧化应激反应

• 批准号: 8168312
• 负责人: JAMES Robert ALFANO
• 金额: $ 7.08万
• 依托单位: UNIVERSITY OF NEBRASKA LINCOLN
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-08-01 至 2011-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8168312
• 关键词: Animal Model; Animals; Antioxidants; Bacillus subtilis; Bacteria; Cells; Complex; Computer Retrieval of Information on Scientific Projects Database; Escherichia coli; Funding; Funding Opportunities; Gene Expression; Genome; Goals; Grant; Homeostasis; Immune response; Immune system; Individual; Institution; Life; Mediating; Metabolic; Molecular; Organism; Oxidation-Reduction; Oxidative Stress; Pathogenicity; Plants; Process; Proteins; Proteomics; Pseudomonas; Pseudomonas aeruginosa; Regulation; Research; Research Personnel; Resources; Role; Signal Transduction; Source; Stress; Stress Tests; System; Systems Biology; United States National Institutes of Health; biological adaptation to stress; biological systems; commensal microbes; pathogen; trait; transcriptomics

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Stress response is the ability of an organism to adjust to drastic changes in environmental parameters. All living organisms have genetically encoded stress response and adaptation systems. Oxidative stress is a common stress form at the cellular level. Recent revolutionary progress in high-throughput experimental and computational approaches offer an opportunity to characterize the molecular mechanisms of oxidative stress response at the level of organisms, cells, genomes, regulatory networks and individual components. We propose to characterize stress response and regulation by focusing on oxidative stress response mechanisms. We will examine Pseudomonas bacterial species under conditions of oxidative stress to (i) identify and quantify gene expression changes, (ii) analyze proteomic and metabolic changes, (iii) build a global transcriptomic and proteomics networks, and (iv) characterize oxidative stress response networks. Much of the proposed research is cutting edge, has not been performed in any biological system, and it will allow for a new avenue of research for the Alfano and Becker research groups and new grant funding opportunities. Research into the mechanisms by which model organisms maintain redox homeostasis have revealed it to be an intricate and complex process. Bacterial antioxidant mechanisms are best understood in commensal Escherichia coli and the Gram-positive model organism Bacillus subtilis (saprophyte) (41), however, there are significant gaps in our understanding of redox homeostasis in non-model organisms. Here we seek to use systems biology approaches to determine if the mechanisms by which bacteria that are exposed to the intense oxidative stress response of the innate immune system vary from that of free-living or commensal bacteria. Specifically, we will use the animal pathogen Pseudomonas aeruginosa and the plant pathogen P. syringae. Both species are exposed to endogenous oxidative stress and exposed to oxidative stress from their host's innate immune response. It will be informative to compare and contrast the importance of oxidative stress responses in pathogenicity of plants and animals. A systems biology approach will allow for a greater understanding of the divergent and convergent evolutionary traits that these bacteria have acquired. We anticipate that we will identify oxidative stress response mechanisms that are common to both species. Our long-term goal is to understand how redox signals from biotic stress are mediated in Gram-negative pathogens of plants and animals to elucidate mechanisms of oxidative stress protection. The Specific Aims of this application are as follows: Identify and quantitate gene expression changes in P. aeruginosa and P. syringae under different oxidative stress conditions; Analyze proteomic and metabolic changes during oxidative stress in P. aeruginosa and P. syringae; Build global network of transcriptomic and proteomic changes induced by oxidative stress; and test role of gene products in oxidative stress response.

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 应激反应是生物体适应环境参数剧烈变化的能力。所有生物体都有基因编码的应激反应和适应系统。氧化应激是细胞水平上常见的应激形式。最近在高通量实验和计算方法的革命性进展提供了一个机会，在生物体，细胞，基因组，调控网络和单个组件的水平上表征氧化应激反应的分子机制。我们建议通过关注氧化应激反应机制来表征应激反应和调节。我们将在氧化应激条件下检查假单胞菌属细菌物种，以（i）识别和量化基因表达变化，（ii）分析蛋白质组学和代谢变化，（iii）建立全球转录组学和蛋白质组学网络，（iv）表征氧化应激反应网络。大部分拟议的研究都是尖端的，尚未在任何生物系统中进行，它将为阿尔法诺和贝克尔研究小组提供新的研究途径和新的赠款资助机会。对模式生物维持氧化还原稳态的机制的研究表明，这是一个错综复杂的过程。细菌抗氧化机制在大肠杆菌和革兰氏阳性模式生物枯草芽孢杆菌（枯草芽孢杆菌）中得到了最好的理解（41），然而，我们对非模式生物中氧化还原稳态的理解存在重大差距。在这里，我们试图使用系统生物学方法来确定是否暴露于先天免疫系统的强烈氧化应激反应的细菌的机制不同于自由生活或寄生细菌。具体来说，我们将使用动物病原体铜绿假单胞菌和植物病原体丁香假单胞菌。这两个物种都暴露于内源性氧化应激和暴露于来自其宿主先天免疫反应的氧化应激。比较和对比氧化应激反应在植物和动物致病性中的重要性将是有益的。系统生物学方法将允许更好地理解这些细菌获得的发散和收敛的进化特征。我们预计，我们将确定氧化应激反应机制是共同的两个物种。我们的长期目标是了解生物胁迫的氧化还原信号是如何在植物和动物的革兰氏阴性病原体中介导的，以阐明氧化应激保护机制。 本申请的具体目的如下：鉴定和定量不同氧化应激条件下铜绿假单胞菌和P. erythingae中的基因表达变化;分析铜绿假单胞菌和P. erythingae中氧化应激期间的蛋白质组学和代谢变化;构建由氧化应激诱导的转录组学和蛋白质组学变化的全球网络;并测试基因产物在氧化应激反应中的作用。