Copper Homeostasis in Escherichia coli

描述：(申请人提供)作为所有生物的基本要素 在电池中，当铜被允许积累超过 手机的需求。细胞内游离铜浓度控制在以下范围内 两种铜外排ATPase的毒性水平 受金属敏感转录因子调控，以及金属配位配位酮 引导铜进入靶蛋白并保护这种高度活性的元素 不受意外行为的伤害。然而，对分子和分子的了解很少 这些过程的机械原理。简单的生物体，如大肠杆菌 Coli为研究细胞内铜的动态平衡提供了一个很好的模型系统。 最近的研究表明，在大肠杆菌中，铜的反应转录 调节者，即CueR，负责主体的基因表达 铜出口商，COPA。这项建议侧重于 CueR介导的CopA基因转录调控及分子和 CueR在金属识别中的结构基础。通过校准铜 CueR/CopA基因相互作用的敏感性 一种简便的测定细胞内游离铜浓度的探针 大肠埃希菌。这些结果可能为实验证据的重要性提供实验证据 原核细胞中的金属配位体，并可能导致发现 第一个在大肠杆菌中发现的铜伴侣。这些研究为以下工作奠定了基础 两种原核生物金属动态平衡的基本原理 和真核细胞。

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.