Copper Homeostasis in Escherichia coli

大肠杆菌中的铜稳态

• 批准号: 6626264
• 负责人: KUI CHEN-HO
• 金额: $ 0.43万
• 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/6626264
• 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 细胞中，铜如果积累过量就会产生剧毒。 细胞需求。细胞内游离铜浓度控制在以下 两种铜流出 ATP 酶的毒性水平，其基因表达为 受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度活跃的元素 来自冒险行动。然而，人们对它的分子和性质知之甚少。 这些过程的机械原理。简单生物，如埃希氏菌 大肠杆菌提供了一个优秀的模型系统来研究细胞内的铜稳态。 最近的研究表明，在大肠杆菌中，铜反应性转录 调节子，即 CueR，负责主要基因的表达 铜出口国，CopA。该提案侧重于机制细节 CueR 介导的 CopA 基因转录调控及其分子和 CueR 在金属识别中的结构基础。通过校准铜 CueR/CopA 基因相互作用的体外敏感性，该提案提供了 一种方便的探针来测定细胞内游离铜的浓度 大肠杆菌。这些结果可能为证明其重要性提供实验证据 原核细胞中的金属伴侣，并可能导致发现 大肠杆菌中的第一个铜伴侣。这些研究奠定了基础 描述原核生物中金属稳态的基本原理 和真核细胞。