Gertz S, Engelmann S, Schmid R, Ziebandt A K, Tischer K, Scharf C, Hacker J, Hecker M
Institut für Mikrobiologie und Molekularbiologie, D-17487 Greifswald, Germany.
J Bacteriol. 2000 Dec;182(24):6983-91. doi: 10.1128/JB.182.24.6983-6991.2000.
The sigma(B)-dependent stress regulon in gram-positive bacteria might fulfill a physiological role in stress response and virulence similar to that of the sigma(S) regulon in Escherichia coli and other gram-negative bacteria. In order to obtain evidence for the function of the sigma(B) regulon of Staphylococcus aureus, especially in virulence control, sigma(B)-dependent stress genes were identified. The two-dimensional protein pattern of wild-type cells of S. aureus COL was compared with that of an isogenic sigB mutant. By this approach, we found that the synthesis of about 27 cytoplasmic proteins seemed to be under the positive control of sigma(B). N-terminal sequencing of 18 proteins allowed the identification of their genes on the almost finished genome sequence of S. aureus COL and the analysis of the promoter structure. Transcriptional analyses of 11 of these genes confirmed their sigma(B) dependency, and moreover, about 7 additional sigma(B)-dependent genes were found which are cotranscribed with the newly detected genes, forming operons. Altogether, we identified 23 sigma(B)-dependent genes and their corresponding proteins. Among them are proteins probably involved in the generation of NADH or in membrane transport mechanisms. Furthermore, at least one clpC-homologous gene was localized on the S. aureus sequence solely transcribed by sigma(B). In contrast, a second clpC-homologous gene in S. aureus forming an operon with ctsR, yacH, and yacI was sigma(B) independently expressed.
革兰氏阳性菌中依赖σ(B)的应激调节子可能在应激反应和毒力方面发挥类似于大肠杆菌及其他革兰氏阴性菌中σ(S)调节子的生理作用。为了获得金黄色葡萄球菌σ(B)调节子功能的证据,尤其是在毒力控制方面的证据,我们鉴定了依赖σ(B)的应激基因。将金黄色葡萄球菌COL野生型细胞的二维蛋白质图谱与同基因sigB突变体的图谱进行了比较。通过这种方法,我们发现约27种细胞质蛋白的合成似乎受σ(B)的正调控。对18种蛋白质进行N端测序,从而在金黄色葡萄球菌COL几乎完成的基因组序列上鉴定出它们的基因,并对启动子结构进行分析。对其中11个基因的转录分析证实了它们对σ(B)的依赖性,此外,还发现了约7个与新检测到的基因共转录的依赖σ(B)的基因,形成了操纵子。我们总共鉴定出23个依赖σ(B)的基因及其相应的蛋白质。其中包括可能参与NADH生成或膜转运机制的蛋白质。此外,在金黄色葡萄球菌序列上至少有一个与clpC同源的基因仅由σ(B)转录。相比之下,金黄色葡萄球菌中与ctsR、yacH和yacI形成操纵子的第二个与clpC同源的基因是独立于σ(B)表达的。