Homerova Dagmar, Bischoff Markus, Dumolin Alexis, Kormanec Jan
Institute of Molecular Biology, Center of Excellence for Molecular Medicine, Slovak Academy of Sciences, Dubravska cesta 21, 845 51 Bratislava, Slovak Republic.
FEMS Microbiol Lett. 2004 Mar 19;232(2):173-9. doi: 10.1016/S0378-1097(04)00063-1.
We optimized a previously established two-plasmid system for the identification of Staphylococcus aureus promoters that are recognized by the alternative transcription factor sigma(B). The method allowed the identification of 18 S. aureus sigma(B)-dependent promoters, 12 of which are reported here for the first time to be sigma(B)-dependent. S1-nuclease mapping of the respective transcriptional start points revealed that all the promoters contained sequences exhibiting high similarity to the consensus sequence of Bacillus subtilis sigma(B)-dependent promoters. The promoters governed expression of genes encoding proteins proposed to be involved in various cellular functions, including the stress response genes and virulence-associated clfA gene for fibrinogen-binding clumping factor. Comparison of the nucleotide sequences upstream of the identified transcription start points identified a sigma(B) consensus promoter (GttTaa-N(12-15)-gGGTAt) that is highly homologous to that of sigma(B) of B. subtilis.
我们优化了先前建立的双质粒系统，用于鉴定由替代转录因子σ(B)识别的金黄色葡萄球菌启动子。该方法能够鉴定出18个金黄色葡萄球菌σ(B)依赖性启动子，其中12个首次被报道为σ(B)依赖性。对各个转录起始点的S1核酸酶作图显示，所有启动子都含有与枯草芽孢杆菌σ(B)依赖性启动子的共有序列高度相似的序列。这些启动子调控着编码参与各种细胞功能的蛋白质的基因的表达，包括应激反应基因和与毒力相关的纤维蛋白原结合聚集因子clfA基因。对已鉴定的转录起始点上游核苷酸序列的比较确定了一个σ(B)共有启动子(GttTaa-N(12 - 15)-gGGTAt)，它与枯草芽孢杆菌的σ(B)高度同源。
