Anaerobic Expression of Fumarate Reductase in E. coli

富马酸还原酶在大肠杆菌中的厌氧表达

• 批准号: 7214841
• 负责人: ROBERT P GUNSALUS
• 金额: $ 36.02万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 1986
• 资助国家: 美国
• 起止时间: 1986-09-01 至 2009-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7214841
• 关键词: Anabolism; Animals; Area; Biochemical; Cell Maintenance; Cells; Complex; DNA; Ensure; Enteral; Enterobacteriaceae; Environment; Enzymes; Escherichia; Escherichia coli; Gene Expression; Gene Family; Generations; Genes; Harvest; Methods; Microarray Analysis; Microbe; Modeling; Molecular; Nitrate Reductases; Nitrates; Nitrite Reductase; Nitrites; Organism; Pathway interactions; Physiology; Plants; Process; Production; Proteins; Reaction; Research Project Grants; Role; Signal Transduction; Succinate Dehydrogenase; Supporting Cell; Tissue-Specific Gene Expression; TorA protein; base; cell growth; environmental change; insight; microorganism; molybdate; nitrate; nitrate reductase; pathogen; respiratory; response

DESCRIPTION (provided by applicant): The major aims of this research project are to further elucidate the regulatory mechanisms required for nitrate- and molybdate-dependent gene expression in Escherichia coli. This enteric bacterium is a model pathogenic and commensal enteric microorganism: it can detect nitrate and a variety of other respiratory substrates in the environment and respond accordingly by synthesizing different respiratory complexes in response to changing environmental signals. This strategy ensures optimal production of energy to support of cell maintenance reactions, biosynthesis, and cell growth. In the studies proposed herein, we will use several model nitrate/nitrite-controlled genes in this organism (e.g., fumarate reductase (frdABCD), NarG and NapF nitrate reductase (narGHJI, and napA) nitrite reductase (nrf and nirB), and DMSO/TMAO reductase (dmsABC) genes to examine how the alternative environmental signals are detected and how the resulting information is then utilized to modulate gene expression. Since these processes are common to many facultative animal and plant pathogens, the proposed molecular and biochemical studies should provide insight concerning how less well understood microbes manage the anaerobic transitions with respect to nitrate. We will examine the basis for nitrate, nitrite, and molybdate-dependent control of gene expression by the Nar and ModE regulatory circuits. We will extend our studies to further examine how the NarX, NarQ, NarP, and NarL proteins detect and transduce signals to modulate gene control. We will examine how the ModE protein provides for the molybdate-dependent control molybdate-containing enzymes to fine-tune expression of the anaerobic respiratory pathway genes. The Nar and Mode gene families will be identified by microarray analysis. Finally, the physiology of nitrate control will be examined by use of continuous culture methods.

描述（由申请人提供）：本研究项目的主要目的是进一步阐明大肠杆菌中硝酸盐和钼酸盐依赖性基因表达所需的调控机制。这种肠道细菌是一种典型的病原共生肠道微生物：它可以检测环境中的硝酸盐和其他多种呼吸底物，并通过合成不同的呼吸复合物来响应变化的环境信号。这种策略确保了能量的最佳生产，以支持细胞维持反应、生物合成和细胞生长。在本文提出的研究中，我们将使用该生物体中几种硝酸盐/亚硝酸盐控制的模型基因(例如富马酸还原酶（frdABCD）， NarG和NapF硝酸盐还原酶（narGHJI和napA）亚硝酸盐还原酶（nrf和nirB）以及DMSO/TMAO还原酶（dmsABC）基因来研究如何检测替代环境信号以及如何利用所得信息来调节基因表达。由于这些过程在许多兼性动物和植物病原体中是常见的，因此提出的分子和生化研究应该提供关于微生物如何管理硝酸盐厌氧转变的见解。我们将研究硝酸盐、亚硝酸盐和钼酸盐依赖于Nar和ModE调控回路的基因表达控制的基础。我们将进一步研究NarX， NarQ， NarP和NarL蛋白如何检测和转导信号以调节基因控制。我们将研究ModE蛋白如何提供钼酸盐依赖性控制含钼酸盐酶来微调厌氧呼吸途径基因的表达。Nar和Mode基因家族将通过微阵列分析进行鉴定。最后，利用连续培养方法研究硝酸盐控制的生理学。