Salunkhe Prabhakar, Töpfer Tanja, Buer Jan, Tümmler Burkhard
Clinical Reseach Group, OE 6710, Hannover Medical School, Carl Neuberg Str. 1, D-30625 Hannover, Germany.
J Bacteriol. 2005 Apr;187(8):2565-72. doi: 10.1128/JB.187.8.2565-2572.2005.
The steady-state response of Pseudomonas aeruginosa to the oxidative-stress-generating agent hydrogen peroxide was analyzed by PAO1 transcriptome profiling. In total, 694, 411, and 237 genes were upregulated and 668, 576, and 468 genes were downregulated in P. aeruginosa strains TB, 892, and PAO1, respectively. The expression profiles of the two variants of the TB clone were significantly more related to each other than the expression profile of either strain was to that of PAO1. Exposure to H(2)O(2) activated by more than 10-fold the expression of the cyoABCD operon, which is key for aerobic respiration, and of oxidative-stress response elements such as the catalase KatB, the alkyl hydroperoxide reductase AhpF, and the thioredoxin reductase 2 operon. Genes for iron and sulfur homeostasis were upregulated. Most enzymes necessary for the conversion of amino acids into the citric acid cycle were globally downregulated at the transcriptional level. Nitrate respiration and arginine fermentation were shut off in the clone TB strains and attenuated in the PAO strain. The transcriptional profiles indicate that the two clone TB strains are more proficient in coping with H(2)O(2)-mediated oxidative stress than the reference strain PAO. According to this data, we recommend study of the transcriptome of strain PAO1 in parallel with those of at least two strains of another clone in order to differentiate common responses from clone- and strain-specific responses and to minimize overinterpretations of microarray data.
通过PAO1转录组分析,研究了铜绿假单胞菌对产生氧化应激的过氧化氢的稳态反应。在铜绿假单胞菌菌株TB、892和PAO1中,分别有694、411和237个基因上调,668、576和468个基因下调。TB克隆的两个变体的表达谱彼此之间的相关性明显高于任何一个菌株与PAO1的表达谱的相关性。暴露于过氧化氢会使有氧呼吸关键的cyoABCD操纵子以及过氧化氢酶KatB、烷基过氧化氢还原酶AhpF和硫氧还蛋白还原酶2操纵子等氧化应激反应元件的表达增加10倍以上。铁和硫稳态相关基因上调。在转录水平上,氨基酸转化为柠檬酸循环所需的大多数酶整体下调。在克隆TB菌株中,硝酸盐呼吸和精氨酸发酵被关闭,在PAO菌株中则减弱。转录谱表明,两个克隆TB菌株比参考菌株PAO更擅长应对过氧化氢介导的氧化应激。根据这些数据,我们建议在研究PAO1菌株转录组的同时,至少平行研究另一克隆的两个菌株的转录组,以便区分共同反应与克隆和菌株特异性反应,并尽量减少对微阵列数据的过度解读。