Copper Homeostasis in Escherichia coli

大肠杆菌中的铜稳态

• 批准号: 6487915
• 负责人: KUI CHEN-HO
• 金额: $ 3.83万
• 依托单位: NORTHWESTERN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-05-01 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/6487915
• 关键词: Escherichia coli; adenosinetriphosphatase; analytical ultracentrifugation; bacterial genetics; bacterial proteins; biological signal transduction; biological transport; chromatography; copper; cytotoxicity; gene environment interaction; gene induction /repression; genetic promoter element; genetic regulation; membrane transport proteins; metal metabolism; microorganism metabolism; molecular chaperones; nucleic acid structure; postdoctoral investigator; protein folding; protein purification; protein structure function; site directed mutagenesis; spectrometry; transcription factor

DESCRIPTION: (provided by applicant) As an essential element to all living cells, copper can be highly toxic when allowed to accumulate in excess of cellular needs. The intracellular free copper concentration is controlled below the toxic level by both copper efflux ATPases whose gene expressions are regulated by metal-sensitive transcription factors, and metallochaperones that guide copper into the target proteins and protect this highly active element from adventious actions. However, little is known about the molecular and mechanistic principles of these processes. Simple organisms such as Escherichia coli provides an excellent model system to study copper homeostasis in cells. Recent studies have shown that in E. coli a copper-responsive transcription regulator, namely CueR, is responsible for gene expression of the principal copper-exporter, CopA. This proposal focuses on the mechanistic details of the CueR-mediated transcription regulation of the CopA gene and the molecular and structural bases of CueR in metal recognition. By calibrating the copper sensitivity of the CueR/CopA gene interaction in vitro, this proposal provides a convenient probe to determine the intracellular free copper concentration in E. coli. These results may provide experimental evidence for the importance of metallochaperones in prokaryotic cells, and may lead to the discovery of the first copper chaperone in E. coli. These studies lay the groundwork for delineating the fundamental principles of metal homeostasis in both prokaryotic and eukaryotic cells.

描述：（由申请人提供）作为所有生活的基本要素 细胞，铜可以是高度有毒的，当允许积累超过 细胞的需要。细胞内游离铜浓度控制在 两种铜外排ATP酶的毒性水平，其基因表达是 由金属敏感转录因子和金属伴侣调节， 引导铜进入目标蛋白质并保护这种高活性元素 从冒险的行动。然而，人们对分子和 这些过程的机械原理。简单的生物体，如埃希氏菌 coli为研究细胞内铜稳态提供了一个很好的模型系统。 最近的研究表明，在E. coliA铜应答转录 调节子，即CueR，负责主要的基因表达 铜出口商该建议侧重于机械的细节， CueR介导的CopA基因的转录调控以及分子和 CueR在金属识别中的结构基础。通过校准铜 体外CueR/CopA基因相互作用的敏感性，该提议提供了 一个方便的探针，以确定细胞内游离铜浓度， E.杆菌这些结果可能提供实验证据的重要性， 金属伴侣蛋白在原核细胞，并可能导致发现的 在E.杆菌这些研究奠定了基础， 描绘金属稳态的基本原则， 和真核细胞。