Jude Florence, Köhler Thilo, Branny Pavel, Perron Karl, Mayer Matthias P, Comte Rachel, van Delden Christian
Department of Genetics and Microbiology, Centre Médical Universitaire, University of Geneva, Geneva, Switzerland.
J Bacteriol. 2003 Jun;185(12):3558-66. doi: 10.1128/JB.185.12.3558-3566.2003.
Pseudomonas aeruginosa controls the secretion of extracellular virulence factors, including rhamnolipids and LasB elastase, by the las and rhl quorum-sensing systems. Here, we mutated the dksA gene of P. aeruginosa by insertion of an Omega-Hg cassette. The mutant displayed growth rates similar to that of the wild type in rich medium but was impaired in growth in defined minimal medium. Production of rhamnolipids and LasB elastase by the dksA mutant was only 4 and 10%, respectively, of wild-type levels. These defects could be partially complemented by introduction of the plasmid-encoded dksA genes from P. aeruginosa or Escherichia coli. In the dksA mutant, the expression of rhlI was increased early during exponential growth, but expression of other quorum-sensing regulator genes-lasR, lasI, and rhlR-was not affected. Although the transcription of the lasB and rhlAB genes was comparable between the dksA mutant and the wild-type strain in peptone tryptic soy broth medium, we observed reduced translation of both genes in the dksA mutant. Similarly, we found that full translation of lasB and rhlAB genes in E. coli also requires the dksA gene. DksA is therefore a novel regulator involved in the posttranscriptional control of extracellular virulence factor production in P. aeruginosa.
铜绿假单胞菌通过las和rhl群体感应系统控制包括鼠李糖脂和LasB弹性蛋白酶在内的细胞外毒力因子的分泌。在此,我们通过插入一个Omega-Hg盒对铜绿假单胞菌的dksA基因进行了突变。该突变体在丰富培养基中的生长速率与野生型相似,但在限定的基本培养基中生长受到损害。dksA突变体产生的鼠李糖脂和LasB弹性蛋白酶分别仅为野生型水平的4%和10%。通过引入来自铜绿假单胞菌或大肠杆菌的质粒编码dksA基因,这些缺陷可以得到部分互补。在dksA突变体中,rhlI的表达在指数生长早期增加,但其他群体感应调节基因lasR、lasI和rhlR的表达不受影响。尽管在蛋白胨胰蛋白酶大豆肉汤培养基中,dksA突变体和野生型菌株之间lasB和rhlAB基因的转录相当,但我们观察到dksA突变体中这两个基因的翻译减少。同样,我们发现大肠杆菌中lasB和rhlAB基因的完全翻译也需要dksA基因。因此,DksA是一种参与铜绿假单胞菌细胞外毒力因子产生转录后控制的新型调节因子。