Sentchilo V, Ravatn R, Werlen C, Zehnder A J B, van der Meer J R
Process of Environmental Microbiology and Molecular Ecotoxicology, Swiss Federal Institute for Environmental Science and Technology, CH 8600 Dübendorf, Switzerland.
J Bacteriol. 2003 Aug;185(15):4530-8. doi: 10.1128/JB.185.15.4530-4538.2003.
An unusual type of gene expression from an integrase promoter was found in cultures of the bacterium Pseudomonas sp. strain B13. The promoter controls expression of the intB13 integrase gene, which is present near the right end of a 105-kb conjugative genomic island (the clc element) encoding catabolism of aromatic compounds. The enzymatic activity of integrase IntB13 is essential for site-specific integration of the clc element into the bacterial host's chromosome. By creating transcription fusions between the intB13 promoter and the gfp gene, we showed that integrase expression in strain B13 was inducible under stationary-phase conditions but, strangely, occurred in only a small proportion of individual bacterial cells rather than equally in the whole population. Integrase expression was significantly stimulated by growing cultures on 3-chlorobenzoate. High cell density, heat shock, osmotic shock, UV irradiation, and treatment with alcohol did not result in measurable integrase expression. The occurrence of the excised form of the clc element and an increase in the rates of clc element transfer in conjugation experiments correlated with the observed induction of the intB13'-gfp fusion in stationary phase and in the presence of 3-chlorobenzoate. This suggested that activation of the intB13 promoter is the first step in stimulation of clc transfer. To our knowledge, this is the first report of a chlorinated compound's stimulating horizontal transfer of the genes encoding its very metabolism.
在假单胞菌属菌株B13的培养物中发现了一种来自整合酶启动子的异常基因表达类型。该启动子控制intB13整合酶基因的表达,该基因位于一个105 kb的接合型基因组岛(clc元件)的右端附近,该基因组岛编码芳香族化合物的分解代谢。整合酶IntB13的酶活性对于clc元件位点特异性整合到细菌宿主染色体中至关重要。通过在intB13启动子和gfp基因之间创建转录融合体,我们发现菌株B13中的整合酶表达在稳定期条件下是可诱导的,但奇怪的是,仅在一小部分单个细菌细胞中发生,而不是在整个群体中均匀发生。在3-氯苯甲酸上培养培养物可显著刺激整合酶表达。高细胞密度、热休克、渗透休克、紫外线照射和酒精处理均未导致可测量的整合酶表达。在接合实验中,clc元件切除形式的出现以及clc元件转移率的增加与在稳定期和存在3-氯苯甲酸的情况下观察到的intB13'-gfp融合体的诱导相关。这表明intB13启动子的激活是刺激clc转移的第一步。据我们所知,这是关于一种氯化化合物刺激其自身代谢相关基因水平转移的首次报道。