CadC，金黄色葡萄球菌质粒pI258镉抗性系统的转录调节蛋白。
CadC, the transcriptional regulatory protein of the cadmium resistance system of Staphylococcus aureus plasmid pI258.
作者信息
Endo G, Silver S
机构信息
Department of Microbiology & Immunology, University of Illinois College of Medicine, Chicago 60612-7344, USA.
出版信息
J Bacteriol. 1995 Aug;177(15):4437-41. doi: 10.1128/jb.177.15.4437-4441.1995.
Abstract
The CadC protein from the cadA cadmium resistance operon of Staphylococcus aureus plasmid pI258 regulates transcription of this system in vitro. The CadC protein was overproduced in Escherichia coli cells and partially purified. Gel shift assays of the proposed cadA operator/promoter region DNA showed specific association with the CadC protein. Control arsenic resistance operator/promoter DNA from the same plasmid was not shifted by the CadC protein. Cd2+, Bi3+, and Pb2+ caused the release of CadC from DNA in gel retardation assays. DNase I footprinting measurements showed that the CadC protein specifically associated with and protected a region of operator/promoter DNA from nucleotide positions -7 to +14 relative to the start point of mRNA synthesis. Runoff transcription assays with the operator/promoter region of DNA (plus the first 69 nucleotides of the cadC gene) and purified E. coli RNA polymerase gave an mRNA product of the predicted size. Added CadC protein inhibited transcription in vitro.
摘要
金黄色葡萄球菌质粒pI258的cadA镉抗性操纵子中的CadC蛋白在体外调节该系统的转录。CadC蛋白在大肠杆菌细胞中过量表达并部分纯化。对拟议的cadA操纵子/启动子区域DNA进行凝胶迁移试验，结果显示其与CadC蛋白存在特异性结合。来自同一质粒的对照抗砷操纵子/启动子DNA未被CadC蛋白迁移。在凝胶阻滞试验中，Cd2+、Bi3+和Pb2+导致CadC从DNA上释放。DNase I足迹测量表明，CadC蛋白与操纵子/启动子DNA的一个区域特异性结合，并保护该区域免受相对于mRNA合成起始点核苷酸位置-7至+14的核酸酶切割。用DNA的操纵子/启动子区域（加上cadC基因的前69个核苷酸）和纯化的大肠杆菌RNA聚合酶进行的径流转录试验产生了预测大小的mRNA产物。添加的CadC蛋白在体外抑制转录。
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