Bernardini M L, Fontaine A, Sansonetti P J
Unité de Pathogénie Microbienne Moléculaire, Institut National de la Santé et de la Recherche Médicale, Institut Pasteur, Paris, France.
J Bacteriol. 1990 Nov;172(11):6274-81. doi: 10.1128/jb.172.11.6274-6281.1990.
In Shigella flexneri, the ompB locus (containing the ompR and envZ genes) was found to modulate expression of the vir genes, which are responsible for invasion of epithelial cells. vir gene expression was markedly enhanced under conditions of high osmolarity (300 mosM), similar to that encountered in tissues both extra- and intracellularly. Two ompB mutants were constructed and tested for virulence and for osmotic regulation of vir genes. An envZ::Tn10 mutant remained invasive, although its virulence was significantly decreased as a result of its inability to survive intracellularly. By using a vir::lac operon fusion, this mutation was shown to decrease beta-galactosidase expression both in low- and high-osmolarity conditions but did not affect vir expression in response to changes in osmolarity. A delta ompB deletion mutant was also constructed via allelic exchange with an in vitro-mutagenized ompB locus of Escherichia coli. This mutation severely impaired virulence and abolished expression of the vir::lac fusion in both low- and high-osmolarity conditions. Therefore, a two-component regulatory system modulates virulence according to environmental conditions. In addition, the mutation affecting a spontaneous avirulent variant of S. flexneri serotype 5, M90T, has been mapped at the ompB locus and was complemented by the cloned E. coli ompB locus. Introduction of the vir::lac fusion into this mutant did not result in the expression of beta-galactosidase (Lac-).
在福氏志贺菌中,发现ompB位点(包含ompR和envZ基因)可调节负责侵袭上皮细胞的vir基因的表达。在高渗透压(300 mosM)条件下,vir基因的表达显著增强,类似于细胞内外组织中遇到的情况。构建了两个ompB突变体,并对其毒力和vir基因的渗透调节进行了测试。一个envZ::Tn10突变体仍具有侵袭性,尽管由于其无法在细胞内存活,其毒力显著降低。通过使用vir::lac操纵子融合,该突变在低渗透压和高渗透压条件下均显示出β-半乳糖苷酶表达降低,但不影响vir基因对渗透压变化的表达。还通过与体外诱变的大肠杆菌ompB位点进行等位基因交换构建了一个ΔompB缺失突变体。该突变严重损害了毒力,并在低渗透压和高渗透压条件下均消除了vir::lac融合的表达。因此,一个双组分调节系统根据环境条件调节毒力。此外,影响福氏志贺菌血清型5的自发无毒变体M90T的突变已定位在ompB位点,并由克隆的大肠杆菌ompB位点互补。将vir::lac融合引入该突变体不会导致β-半乳糖苷酶的表达(Lac-)。