Brzostek Katarzyna, Brzóstkowska Marta, Bukowska Iwona, Karwicka Ewa, Raczkowska Adrianna
Department of Applied Microbiology, Institute of Microbiology, Warsaw University, Miecznikowa 1, 02-096 Warsaw, Poland.
Microbiology (Reading). 2007 Aug;153(Pt 8):2416-2425. doi: 10.1099/mic.0.2006/003202-0.
Invasin, the major adhesion and invasion factor of Yersinia enterocolitica, is encoded by the inv gene, which is regulated by growth phase and in response to a variety of environmental conditions such as temperature, pH and osmolarity. So far, three proteins, RovA, H-NS and YmoA, have been identified as factors regulating the expression of the inv gene in enteropathogenic Yersinia. Here, data from inv' : : lacZYA chromosomal gene fusion studies are presented indicating that OmpR, the response regulator of the EnvZ/OmpR two-component system, acts to negatively regulate inv expression at the transcriptional level at 25 degrees C, and that high osmolarity enhances the inhibitory effect of this protein. In a strain lacking OmpR the expression of inv at 25 degrees C was increased sixfold, but at 37 degrees C, a temperature known to repress inv expression, this effect was not observed, suggesting that temperature regulation of inv is OmpR-independent. Furthermore, the expression of inv in the ompR background was no longer responsive to increased osmolarity. Complementation with the active ompR allele restored wild-type inv expression in the ompR mutant. In silico analysis of the Y. enterocolitica O : 9 inv promoter sequence revealed the presence of an OmpR consensus binding site located in the -15 to -33 region. OmpR was able to specifically bind to a fragment of the inv promoter containing this putative binding site in electrophoretic mobility shift assays. Thus, OmpR seems to be a repressor of inv in Y. enterocolitica.
侵袭素是小肠结肠炎耶尔森菌的主要黏附与侵袭因子,由inv基因编码,该基因受生长阶段调控,并对多种环境条件作出反应,如温度、pH值和渗透压。到目前为止,已鉴定出三种蛋白质,即RovA、H-NS和YmoA,它们是调节致病性耶尔森菌中inv基因表达的因子。在此,我们展示了来自inv'::lacZYA染色体基因融合研究的数据,表明EnvZ/OmpR双组分系统的反应调节因子OmpR在25℃时在转录水平上对inv表达起负调控作用,并且高渗透压增强了该蛋白的抑制作用。在缺乏OmpR的菌株中,inv在25℃时的表达增加了6倍,但在37℃(已知该温度会抑制inv表达)时未观察到这种效应,这表明inv的温度调节不依赖于OmpR。此外,在ompR背景下inv的表达不再对渗透压升高作出反应。用活性ompR等位基因进行互补可恢复ompR突变体中的野生型inv表达。对小肠结肠炎耶尔森菌O:9 inv启动子序列的计算机分析揭示,在-15至-33区域存在一个OmpR共有结合位点。在电泳迁移率变动分析中,OmpR能够特异性结合包含该假定结合位点的inv启动子片段。因此,OmpR似乎是小肠结肠炎耶尔森菌中inv的一个阻遏物。