Regulation of glycerol utilization in Borrelia burgdorferi

项目总结/摘要 莱姆病是由伯氏疏螺旋体感染引起的。B。burgdorferi是维持在 在包括蜱媒介和脊椎动物宿主的动物流行周期中的自然界。螺旋体转移了它的 碳和能量从脊椎动物的葡萄糖到蜱虫的甘油：甘油的利用是一个关键 这种适应使螺旋体能够在蜱虫中持续存在并成功地传播给脊椎动物。我们假设 甘油代谢和糖酵解的界面，通过甘油-3-磷酸和 二羟丙酮磷酸盐，是导航地方性流行病循环的代谢关键。核心假设 这个应用程序是B。burgdorferi转录后调节甘油代谢的表达 (glp)操纵子利用甘油以便在其蜱虫载体中存活。我们奋进剖析 调节glp操纵子表达的机制，包括一种新的反义RNA的特殊作用， 环状二聚GMP（c-di-GMP）结合蛋白PlzA。提出了以下具体目标， 实现这一目标：1）确定参与甘油代谢的基因在地方性流行病中的作用， 周期; 2）确定glp操纵子调节的分子机制;和3）确定PlzA和 确定其在调节GLP操纵子中的作用。遗传、生物化学、分子和转录组学 方法，以及莱姆病的蜱鼠模型，将用于实验测试这些 假设在这个创新的应用程序中，多个PI处于独特的位置，拥有经验， 专业知识和实验模型，以加强对碳水化合物代谢机制的理解 以及在莱姆病病原体在其蜱媒中持续存在期间的调节。的长期目标 这个建议是为了理解允许B的调节机制。burgdorferi回应和适应 在蜱虫到哺乳动物的传播和莱姆病的发病机制中， 改善诊断，预防和治疗策略，因为甘油和葡萄糖的利用率 在蜱虫和脊椎动物感染期间分别对持久性至关重要;这与 为了减轻人类疾病，该机构“寻求基础知识”。

PROJECT SUMMARY/ABSTRACT Lyme disease is caused by infection with the spirochete Borrelia burgdorferi. B. burgdorferi is maintained in nature in an enzootic cycle comprising a tick vector and vertebrate host. The spirochete shifts its source of carbon and energy from glucose in the vertebrate to glycerol in the tick: utilization of glycerol is a crucial adaptation for the spirochete to persist in the tick and successfully transmit to a vertebrate. We hypothesize that the interface of glycerol metabolism and glycolysis, linked by interconversion of glycerol-3-phosphate and dihydroxyacetone phosphate, is the metabolic linchpin for navigating the enzootic cycle. The central hypothesis of this application is that B. burgdorferi post-transcriptionally regulates expression of the glycerol metabolism (glp) operon to utilize glycerol in order to survive in its tick vector. We endeavor to dissect the molecular mechanisms regulating expression of the glp operon, including the specific roles of a novel antisense RNA and the cyclic-dimeric GMP (c-di-GMP)-binding protein PlzA. The following specific aims are proposed toward achieving this objective: 1) define the role of the genes involved in glycerol metabolism during the enzootic cycle; 2) determine the molecular mechanism of glp operon regulation; and 3) define the function of PlzA and determine its role in regulating the glp operon. Genetic, biochemical, molecular, and transcriptomic approaches, as well as the tick-mouse model of Lyme disease, will be utilized to experimentally test these hypotheses. In this innovative application, the Multiple-PIs are in the unique position of having the experience, expertise, and experimental models to enhance the mechanistic understanding of carbohydrate metabolism and its regulation during the persistence of the Lyme disease agent in its tick vector. The long-term objective of this proposal is to understand the regulatory mechanisms that allow B. burgdorferi to respond and adapt to varied carbon sources in the tick-to-mammal transmission and in Lyme disease pathogenesis, which will lead to improved diagnostic, prevention, and treatment strategies because glycerol and glucose utilization are critical for persistence during tick and vertebrate infection, respectively; this is relevant to the mission of the agency to “seek fundamental knowledge” for the sake of alleviating human disease.