Genetic Mechanisms in Borrelia burgdorferi Pathogenesis

伯氏疏螺旋体发病机制的遗传机制

• 批准号: 7589384
• 负责人: JON T SKARE
• 金额: $ 36.28万
• 依托单位: TEXAS A&M UNIVERSITY HEALTH SCIENCE CTR
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-04-01 至 2014-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7589384
• 关键词: Address; Affect; Arthropod Vectors; Arthropods; Avidity; Bacteria; Binding; Biochemical; Biology; Blood; Borrelia; Borrelia burgdorferi; Borrelia oxidative stress regulator; Case Study; Cells; Centers for Disease Control and Prevention (U.S.); Development; Disease; Drug Metabolic Detoxication; Emerging Communicable Diseases; Environment; Essential Genes; Funding; Gene Expression; Genes; Genetic; Goals; Homeostasis; Immune response; In Vitro; Infectious Agent; Isopropyl Thiogalactoside; Knock-out; Knowledge; Life Cycle Stages; Link; Lipase; Lyme Disease; Mammals; Mediating; Metals; Modeling; Modification; Molecular; Molecular Genetics; Morbidity - disease rate; Operon; Order Spirochaetales; Organism; Oxidation-Reduction; Oxidative Stress; Pathogenesis; Patients; Phospholipids; Physiological; Physiology; Process; Protein-Serine-Threonine Kinases; Proteins; Public Health; Reactive Oxygen Species; Regulation; Relative (related person); Research Personnel; Role; Signal Transduction; Staging; Structural Models; System; Temperature; Testing; Ticks; Transcript; United States; Virulence; Virulent; Work; acyl group; biological adaptation to stress; combat; design; expectation; genetic analysis; genetic regulatory protein; genome sequencing; innovation; insight; mutant; oxidation; pathogen; pathogenic bacteria; polyunsaturated fat; promoter; public health relevance; response; trafficking; transmission process; vector

DESCRIPTION (provided by applicant): Borrelia burgdorferi, the etiologic agent of Lyme disease, is the most common arthropod- borne infectious agent in the United States, and contributes to a significant amount of morbidity in persistently infected patients. B. burgdorferi is effective at colonizing both mammalian and arthropod hosts and, as such, must modulate gene expression quickly to adapt to these different environments. Although we know some of the molecular signals that alter gene expression in B. burgdorferi, we still understand little regarding how potential virulence determinants are regulated in this pathogen. In the past funding period, we have characterized a regulatory protein, designated BosR, which is involved in regulating the oxidative stress response in B. burgdorferi. Recently, we found that two genes linked to bosR, bb0646 and bb0648, share a transcript and thus comprise an operon. These genes encode for an exported lipase (bb0646) and a serine/threonine kinase (bb0648), respectively, which we suggest are involved in the oxidative stress response in B. burgdorferi. The central hypothesis is that BosR, and the linked genes bb0646 and bb0648, coordinate an important adaptive response that senses the redox status of the cell. To address this hypothesis, we propose the following Specific Aims: (1) Characterize the bosR operon in infectious B. burgdorferi. The working hypothesis is that bosR and its flanking genes, bb0646 and bb0648, respond appropriately to the redox status of the cell to combat toxic oxidizing compounds generated during the arthropod blood meal or the mammalian innate immune response. We have not yet been able to evaluate the role of BosR in infectious isolates, presumably since bosR regulates essential genes. Here we will use a recently developed tightly regulated inducible system to generate a conditional mutant in bosR in infectious B. burgdorferi; (2) Assess the infectivity deficit in conditional mutants and knockouts in BosR-regulated genes. The working hypothesis is that genes regulated by BosR are required for physiologically important processes related to oxidative stress and infectivity; (3) Determine the mechanism of BosR-mediated regulation. The working hypothesis is that BosR alters its regulatory activity via oxidation and metal binding, which changes its avidity for target sequences; and (4) Decipher the role of BB0646 and BB0648 in B. burgdorferi pathogenesis. Our working hypothesis is that both of these gene products are involved in host adaptation by modifying polyunsaturated lipid substrates and coordinating a global response to oxidative stress, respectively. The information from these studies will provide insight into how B. burgdorferi adapts to the redox status of the host via BosR, BB0646, and BB0648, and will help to determine how the ensuing response relates to the disease potential of this important pathogen. PUBLIC HEALTH RELEVANCE: Borrelia burgdorferi, the etiologic agent of Lyme disease, is the most common arthropod-borne infectious agent in the United States, and thus is an important Public Health issue. The studies described herein are designed to address how B. burgdorferi is able to adapt to both ticks and mammals, in the context of host mediated oxidation defenses, and how this adaptive response affects the ability of this bacterium to persist within the environment and cause disease.

描述（由申请方提供）：莱姆病的病原体伯氏疏螺旋体是美国最常见的节肢动物传播的传染性病原体，并导致持续感染患者的大量发病率。B。Burgdorferi在哺乳动物和节肢动物宿主中都是有效的，因此必须快速调节基因表达以适应这些不同的环境。虽然我们知道一些改变B基因表达的分子信号。尽管存在burgdorferi，但我们对这种病原体中潜在毒力决定因素如何调节仍然知之甚少。在过去的资助期间，我们已经确定了一种调节蛋白，命名为BosR，它参与调节B中的氧化应激反应。burgdorferi。最近，我们发现与bosR相关的两个基因bb 0646和bb 0648共享一个转录本，因此包含一个操纵子。这些基因编码的出口脂肪酶（bb 0646）和丝氨酸/苏氨酸激酶（bb 0648），分别，我们建议参与氧化应激反应在B。burgdorferi。核心假设是BosR和相关基因bb 0646和bb 0648协调了一种重要的适应性反应，这种反应可以感知细胞的氧化还原状态。为了解决这一假设，我们提出了以下具体目标：（1）表征传染性B中的bosR操纵子。burgdorferi。工作假设是，bosR及其侧翼基因，bb 0646和bb 0648，适当地响应细胞的氧化还原状态，以对抗节肢动物血餐或哺乳动物先天免疫反应过程中产生的有毒氧化化合物。我们尚未能够评估BosR在感染性分离株中的作用，大概是因为BosR调节必需基因。在这里，我们将使用一个最近开发的严格监管的诱导系统，以产生一个有条件的突变体在感染性B的bosR。（2）评估条件突变体的感染性缺陷和BosR调控基因的敲除。工作假设：BosR调控的基因是与氧化应激和感染性相关的重要生理过程所必需的;（3）确定BosR介导的调控机制。工作假设是BosR通过氧化和金属结合改变其调节活性，这改变了其对靶序列的亲合力;和（4）解密BB 0646和BB 0648在B中的作用。burgdorferi发病机制我们的工作假设是，这两种基因产物分别通过修饰多不饱和脂质底物和协调对氧化应激的全球反应参与宿主适应。这些研究的信息将提供对B。Burgdorferi通过BosR，BB 0646和BB 0648适应宿主的氧化还原状态，并将有助于确定随后的反应如何与这种重要病原体的疾病潜力相关。公共卫生相关性：莱姆病的病原体伯氏疏螺旋体（Borrelia burgdorferi）是美国最常见的节肢动物传播的传染性病原体，因此是重要的公共卫生问题。本文所述的研究旨在解决如何B。在宿主介导的氧化防御的背景下，伯氏菌能够适应蜱和哺乳动物，以及这种适应性反应如何影响这种细菌在环境中持续存在并引起疾病的能力。