Molecular Genetics Group, Groningen Biomolecular Sciences and Biotechnology Institute, University of Groningen, Groningen, The Netherlands.
PLoS One. 2013;8(1):e53471. doi: 10.1371/journal.pone.0053471. Epub 2013 Jan 3.
The genes for DNA uptake and recombination in Bacilli are commonly regulated by the transcriptional factor ComK. We have identified a ComK homologue in Bacillus coagulans, an industrial relevant organism that is recalcitrant for transformation. Introduction of B. coagulans comK gene under its own promoter region into Bacillus subtilis comK strain results in low transcriptional induction of the late competence gene comGA, but lacking bistable expression. The promoter regions of B. coagulans comK and the comGA genes are recognized in B. subtilis and expression from these promoters is activated by B. subtilis ComK. Purified ComK protein of B. coagulans showed DNA-binding ability in gel retardation assays with B. subtilis- and B. coagulans-derived probes. These experiments suggest that the function of B. coagulans ComK is similar to that of ComK of B. subtilis. When its own comK is overexpressed in B. coagulans the comGA gene expression increases 40-fold, while the expression of another late competence gene, comC is not elevated and no reproducible DNA-uptake could be observed under these conditions. Our results demonstrate that B. coagulans ComK can recognize several B.subtilis comK-responsive elements, and vice versa, but indicate that the activation of the transcription of complete sets of genes coding for a putative DNA uptake apparatus in B. coagulans might differ from that of B. subtilis.
芽孢杆菌中的 DNA 摄取和重组基因通常受转录因子 ComK 调控。我们在凝结芽孢杆菌(一种工业上相关的转化抗性生物体)中鉴定出了 ComK 同源物。在芽孢杆菌枯草亚种的 comK 菌株中,引入凝结芽孢杆菌 comK 基因及其自身启动子区域,导致晚期感受态基因 comGA 的转录诱导水平较低,但缺乏双稳态表达。凝结芽孢杆菌 comK 和 comGA 基因的启动子区域在枯草芽孢杆菌中被识别,并且这些启动子的表达被枯草芽孢杆菌 ComK 激活。凝结芽孢杆菌的 ComK 蛋白在凝胶阻滞实验中显示出与枯草芽孢杆菌和凝结芽孢杆菌衍生探针的 DNA 结合能力。这些实验表明,凝结芽孢杆菌 ComK 的功能类似于枯草芽孢杆菌的 ComK。当凝结芽孢杆菌自身的 comK 过表达时,comGA 基因的表达增加了 40 倍,而另一个晚期感受态基因 comC 的表达并没有升高,并且在这些条件下没有观察到可重复的 DNA 摄取。我们的结果表明,凝结芽孢杆菌 ComK 可以识别几个枯草芽孢杆菌 comK 响应元件,反之亦然,但表明凝结芽孢杆菌中完整的一组编码假定 DNA 摄取装置的基因的转录激活可能与枯草芽孢杆菌不同。