Molecular mechanism of general stress signaling in Brucella abortus

流产布鲁氏菌一般应激信号传导的分子机制

• 批准号: 8694631
• 负责人: Sean Crosson
• 金额: $ 43.98万
• 依托单位: UNIVERSITY OF CHICAGO
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-02-01 至 2019-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8694631
• 关键词: Address; Affect; Affinity; Agriculture; Animals; Bacteria; Binding; Binding Sites; Biochemical; Biochemistry; Biological; Biophysics; Brucella; Brucella abortus; Brucellosis; Cells; Cellular biology; Chemicals; Chicago; Chronic; Chronic Phase; Computing Methodologies; Containment; Cues; Data; Detection; Disease; Environment; Flavins; Gene Expression; Genetic; Genetic Transcription; Goals; Growth; Harvest; Health; Human; In Vitro; Infection; Investigation; Laboratories; Measures; Modeling; Molecular; Mus; N-terminal; Peptide Hydrolases; Phosphorylation; Phosphotransferases; Positioning Attribute; Postdoctoral Fellow; Proteins; Proteobacteria; Proteolysis; Regulation; Research; Resistance; Role; Sensory; Signal Pathway; Signal Transduction; Stress; Stress Response Signaling; Structure; System; Testing; Tissues; Transcriptional Activation; Transcriptional Regulation; Universities; Virulence; Work; base; biodefense; biological adaptation to stress; experience; genetic regulatory protein; global health; in vivo; novel; pathogen; pathogenic bacteria; programs; protein function; protein-histidine kinase; public health relevance; reconstitution; release factor; research study; residence; response; sensor; sensor histidine kinase; spatiotemporal; stressor; yeast two hybrid system

DESCRIPTION (provided by applicant): The goal of this research program is to develop a mechanistic understanding of the general stress response (GSR) signaling system in the select agent pathogen, Brucella abortus. Our preliminary data demonstrate that this signaling system controls environmental stress adaptation and chronic infection in a mammalian infection model. B. abortus GSR signaling is controlled in part by the PhyR protein, a conserved regulator of adaptation to environmental stress that is broadly conserved in the ¿-proteobacterial clade. PhyR contains an amino terminal ECF ¿-like domain and a carboxy-terminal TCS receiver domain. Phosphorylation activates PhyR as an anti-anti-¿ factor and releases the classical ECF ¿ factor, ¿E1 to directly regulate gene expression during stress. Our multi-disciplinary investigation of PhyR, ¿E1 and other components of the GSR signaling system will define the mechanistic underpinnings of general stress signaling in a pathogen of significant human health, agricultural, and biodefense import. Moreover, the experiments proposed herein have the potential to inform new treatments for ¿-proteobacterial disease such as brucellosis. As mentioned above, this proposal is highly multi-disciplinary, spanning genetics, biochemistry, molecular biophysics, cell biology, and whole-animal infection studies. I will conduct these studies with a team of three experienced postdoctoral fellows, all of whom have expertise working on this BSL3 select agent in containment at the University of Chicago Howard Taylor Ricketts Regional Biocontainment Laboratory. The specific aims of this project are: 1) Define the role of the sensor histidine kinase, LovhK, as a GSR stress sensor, and characterize the molecular basis of stress signal detection by LovhK. 2) Define signals that activate the B. abortus general stress response in vitro and in a mammalian infection model. 3) Elucidate the structural mechanism of GSR activation.

描述（由申请人提供）：本研究计划的目标是对选择病原体布鲁氏菌（Brucella abortus）的一般应激反应（GSR）信号系统进行机制理解。我们的初步数据表明，在哺乳动物感染模型中，该信号系统控制着环境应激适应和慢性感染。B. abortus的GSR信号部分由PhyR蛋白控制，PhyR蛋白是一种保守的适应环境压力的调节剂，在蛋白质细菌分支中广泛保守。PhyR包含一个氨基端ECF¿样结构域和一个羧基端TCS接收器结构域。磷酸化激活PhyR作为抗抗因子，释放经典的ECF因子E1，直接调节应激过程中的基因表达。我们对PhyR，¿E1和GSR信号系统的其他组成部分的多学科研究将确定对人类健康，农业和生物防御具有重要意义的病原体中一般应激信号的机制基础。此外，本文提出的实验有可能为布鲁氏菌病等变形菌疾病的新治疗提供信息。如上所述，该方案是高度跨学科的，涵盖遗传学、生物化学、分子生物物理学、细胞生物学和全动物感染研究。我将与一个由三名经验丰富的博士后组成的团队一起进行这些研究，他们都在芝加哥大学霍华德·泰勒·里基茨地区生物防护实验室从事BSL3筛选剂的专业工作。本课题的具体目的是：1)明确传感器组氨酸激酶LovhK作为GSR应激传感器的作用，并表征LovhK检测应激信号的分子基础。2)在体外和哺乳动物感染模型中定义激活B. abortus一般应激反应的信号。3)阐明GSR活化的结构机制。