Oxidative and Nitrosative Stress in Burkholderia

本项目的目标是确定适应性反应的主要分子成分。 假马利氏伯克霍尔德氏菌的活性氧和氮胁迫。假腮腺杆菌是病原菌 类鼻疽的症状。高死亡率，困难的治疗方案，以及广泛的致病性 菌株导致假鼻疽杆菌被归类为潜在的生物恐怖主义病原体。更有效 需要治疗方法和有效的疫苗来防御假鼻疽的攻击。在.期间 在感染过程中，细胞内的病原体必须应对各种宿主介导的应激条件，在 特别是吞噬细胞的抗菌特性。B.假腮腺杆菌入侵并顽固存在 巨噬细胞。巨噬细胞产生抗微生物的活性氧和氮(ROS和RNS)。 细胞内病原体躲避这些抗菌剂的机制是它们生存的核心 在感染期间。假鼻疽杆菌基因组中与ROS抗性有关的基因 包括oxR和soxR基因，以及超氧化物歧化酶、过氧化氢酶和烷基基因 过氧化氢还原酶。我们的工作假设是：1)允许假单胞菌 抵抗ROS和RN的杀伤对于疾病期间的生存至关重要，因此是合乎逻辑的候选 抗微生物干预和2)耐药蛋白在疾病期间高度表达，因此是理想的 疫苗研发的候选对象。我们将研究假单胞菌对氧化和亚硝酸盐的适应 应激使用在我们的结核分枝杆菌研究中改进的全局表达谱方法。我们 将鉴定假鼻疽杆菌对ROS和RNS的转录反应，确定主要的 氧化应激调节剂(OxyR和SoxR)在防御ROS和RNS中的作用，并确定关键的抗性 用于疫苗和药物开发的蛋白质。除了中介绍的交互之外的项目交互 我们将与其他项目成员(例如，Vasil博士、Holmes博士和Vazquez-Torres博士)进行互动 Slayden博士(基因组学/蛋白质组学核心-CSU)在开发和使用微阵列技术和 治疗药物潜在靶蛋白的评估，罗宾逊博士(选择试剂档案-BYU) 获取具有代表性的假鼻疽杆菌分离株，以及Schweizer博士(微生物遗传学-CSU) 假鼻疽杆菌的遗传学研究。

Introduction The goal of this project is to identify the major molecular components of the adaptive response of Burkholderia pseudomalleiio reactive oxygen and nitrogen stress. B. pseudomallei is the causative agent of melioidosis. High mortality rates, difficult treatment regimens, and the wide availability of pathogenic strains have led to the classification of B. pseudomallei as a potential bioterrorism agent. More effective treatments and an efficacious vaccine are needed to defend against a B. pseudomallei attack. During the course of infection, intracellular pathogens must cope with a variety of host-mediated stress conditions, in particular, the antibacterial properties of phagocytic cells. B. pseudomallei invades and persists in macrophages. Macrophages produce antimicrobial reactive oxygen and nitrogen species (ROS and RNS). Mechanisms by which intracellular pathogens avoid these antibacterial agents are central to their survival during infection. Genes involved in resistance to ROS that are present in the B. pseudomallei genome include the oxyR and soxR genes, and the genes for superoxide dismutases, catalases, and alkyl hydroperoxide reductases. Our working hypotheses are: 1) mechanisms which allow B. pseudomallei to resist killing by ROS and RNS are critical for survival during disease and are thus logical candidates for antimicrobial intervention and 2) resistance proteins are highly expressed during disease and thus ideal candidates for vaccine development. We will study B. pseudomallei's adaptation to oxidative and nitrosative stress using global expression profiling methods refined in our studies of Mycobacterium tuberculosis. We will identify the transcriptional response of B. pseudomallei to ROS and RNS, determine the role of the major oxidative stress regulators (OxyR and SoxR) in defense against ROS and RNS, and identify key resistance proteins for vaccine and drug development. Project interactions In addition to the interactions described in this proposal with other project members (e.g. Drs. Vasil, Holmes, & Vazquez-Torres), we will interact with Dr. Slayden (Genomics/Proteomics Core-CSU) in the development and use of microarray technology and the evaluation of potential target proteins for therapeutic agents, Dr. Robinson (Select Agent Archive-BYU) for access to representative B. pseudomallei isolates, and Dr. Schweizer (Microbial Genetics-CSU) in terms of genetics of B. pseudomallei.