Prince R W, Storey D G, Vasil A I, Vasil M L
Department of Microbiology and Immunology, University of Colorado Health Sciences Center, Denver 80262.
Mol Microbiol. 1991 Nov;5(11):2823-31. doi: 10.1111/j.1365-2958.1991.tb01991.x.
A multicopy plasmid containing the Escherichia coli fur gene was introduced into Pseudomonas aeruginosa strain PA103C. This strain contains a toxA-lacZ fusion integrated into its chromosome at the toxA locus. Beta-galactosidase synthesis in this strain is regulated by iron, as is seen for exotoxin A production. Beta-galactosidase synthesis and exotoxin A production in PA103C containing multiple copies of E. coli fur was still repressed in low iron conditions. The transcription of regA, a positive regulator of toxA, was also found to be inhibited by multiple copies of the E. coli fur gene. In addition, the ability of PA103C containing multiple copies of E. coli fur to produce protease was greatly reduced relative to PA103C containing a vector control. A polyclonal rabbit serum containing antibodies that recognize E. coli Fur was used to screen whole-cell extracts from Vibrio cholerae, Shigella flexneri, Salmonella typhimurium and Pseudomonas aeruginosa. All strains tested expressed a protein that was specifically recognized by the anti-Fur serum. These results and those described above suggest that Fur structure and function are conserved in a variety of distinct bacterial genera and that at least some of these different genera use this regulatory protein to control genes encoding virulence factors.
将含有大肠杆菌fur基因的多拷贝质粒导入铜绿假单胞菌PA103C菌株。该菌株含有一个整合到其染色体toxA位点的toxA - lacZ融合基因。该菌株中β - 半乳糖苷酶的合成受铁调控,外毒素A的产生也是如此。在低铁条件下,含有多个大肠杆菌fur拷贝的PA103C中β - 半乳糖苷酶的合成和外毒素A的产生仍受到抑制。toxA的正调控因子regA的转录也被发现受到多个大肠杆菌fur基因拷贝的抑制。此外,相对于含有载体对照的PA103C,含有多个大肠杆菌fur拷贝的PA103C产生蛋白酶的能力大大降低。使用含有识别大肠杆菌Fur抗体的兔多克隆血清筛选霍乱弧菌、福氏志贺菌、鼠伤寒沙门氏菌和铜绿假单胞菌的全细胞提取物。所有测试菌株都表达一种能被抗Fur血清特异性识别的蛋白质。这些结果以及上述结果表明,Fur的结构和功能在多种不同的细菌属中是保守的,并且这些不同属中的至少一些利用这种调节蛋白来控制编码毒力因子的基因。