Liljeström P, Laamanen I, Palva E T
Department of Molecular Biology, Karolinska Institute, Huddinge University Hospital, Sweden.
J Mol Biol. 1988 Jun 20;201(4):663-73. doi: 10.1016/0022-2836(88)90465-2.
The ompB operon of Salmonella typhimurium encodes a positive transcriptional regulator OmpR and an inner membrane protein EnvZ. Both proteins are needed for the proper expression of the outer membrane proteins OmpC and OmpF. We have determined the nucleotide sequence of the ompB locus and its adjacent regions. A comparison between the S. typhimurium and Escherichia coli sequences revealed that the ompB locus is highly conserved. The sequence data also showed that ompR and envZ form an operon, where the coding regions overlap by four base-pairs. Utilizing ompR-lacZ and envZ-lacZ gene fusions, the translational levels of expression of these two genes were measured, showing that ompR is considerably more efficiently expressed than envZ. Analysis of ompR frameshift mutations showed that translation of envZ is almost totally dependent on the translation of the upstream gene ompR. The mechanism of this translational coupling appears to be a reinitiation of the ribosome at the overlapping region of the two genes. The characteristics of the OmpR and EnvZ proteins were in agreement with the known functions and cellular locations of these proteins. OmpR was found to contain a putative DNA binding site, while EnvZ contained two hydrophobic stretches typical of transmembrane regions. Both OmpR and EnvZ show extensive homologies with many proteins from a number of different origins, all of which function in pairs and through which environmental signals modulate gene expression. Hence, the tightly coupled synthesis of these proteins seems to be essential in eliciting a proper response in the transmembrane regulation of gene expression.
鼠伤寒沙门氏菌的ompB操纵子编码一种正向转录调节因子OmpR和一种内膜蛋白EnvZ。这两种蛋白都是外膜蛋白OmpC和OmpF正常表达所必需的。我们已经确定了ompB基因座及其相邻区域的核苷酸序列。鼠伤寒沙门氏菌和大肠杆菌序列的比较显示,ompB基因座高度保守。序列数据还表明,ompR和envZ形成一个操纵子,其编码区域重叠四个碱基对。利用ompR-lacZ和envZ-lacZ基因融合体,测量了这两个基因的翻译水平,结果表明ompR的表达效率比envZ高得多。对ompR移码突变的分析表明,envZ的翻译几乎完全依赖于上游基因ompR的翻译。这种翻译偶联的机制似乎是核糖体在两个基因的重叠区域重新起始。OmpR和EnvZ蛋白的特性与这些蛋白已知的功能和细胞定位一致。发现OmpR含有一个假定的DNA结合位点,而EnvZ含有两个典型的跨膜区域疏水片段。OmpR和EnvZ都与许多来自不同来源的蛋白具有广泛的同源性,所有这些蛋白都是成对发挥作用的,环境信号通过它们来调节基因表达。因此,这些蛋白紧密偶联的合成似乎对于在基因表达的跨膜调节中引发适当的反应至关重要。