Mills S D, Jasalavich C A, Cooksey D A
Department of Plant Pathology, University of California, Riverside 92521-0122.
J Bacteriol. 1993 Mar;175(6):1656-64. doi: 10.1128/jb.175.6.1656-1664.1993.
Specific induction of the copper resistance operon (cop) promoter from Pseudomonas syringae was measured by beta-galactosidase production from a cop promoter-lacZ fusion. Induction of the cop promoter in P. syringae pv. syringae required trans-acting factors from copper resistance plasmid pPT23D, from which cop was originally cloned. Tn5 mutagenesis of pPT23D was used to localize two complementation groups immediately downstream from copABCD. Cloning and sequencing of the DNA in this region revealed two genes, copR and copS, expressed in the same orientation as the cop operon but from a separate constitutive promoter. The amino acid sequence deduced from these genes showed distinct similarities to known two-component regulatory systems, including PhoB-PhoR and OmpR-EnvZ. In addition, CopR showed strong similarity to copper resistance activator protein PcoR from Escherichia coli. Functional chromosomal homologs to copRS activated the cop promoter, in a copper-inducible manner, in copper-resistant or -sensitive strains of P. syringae pv. tomato and other Pseudomonas species. This implies that copper-inducible gene regulation is associated with a common chromosomally encoded function, as well as plasmid-borne copper resistance, in Pseudomonas spp.
通过铜抗性操纵子(cop)启动子与lacZ的融合体产生β-半乳糖苷酶,来测定丁香假单胞菌铜抗性操纵子(cop)启动子的特异性诱导情况。丁香假单胞菌丁香致病变种中cop启动子的诱导需要来自铜抗性质粒pPT23D的反式作用因子,cop最初就是从该质粒中克隆得到的。利用pPT23D的Tn5诱变来定位copABCD下游紧邻的两个互补组。对该区域的DNA进行克隆和测序,发现了两个基因,copR和copS,它们与cop操纵子的方向相同,但来自一个独立的组成型启动子。从这些基因推导的氨基酸序列与已知的双组分调节系统,包括PhoB-PhoR和OmpR-EnvZ,有明显的相似性。此外,CopR与大肠杆菌的铜抗性激活蛋白PcoR有很强的相似性。copRS的功能性染色体同源物能以铜诱导的方式激活丁香假单胞菌番茄致病变种以及其他假单胞菌属的铜抗性或铜敏感菌株中的cop启动子。这意味着在假单胞菌属中,铜诱导基因调控与一种常见的染色体编码功能以及质粒携带的铜抗性相关。