Mazurakova Vladislava, Sevcikova Beatrica, Rezuchova Bronislava, Kormanec Jan
Institute of Molecular Biology, Center of Excellence for Molecular Medicine, Slovak Academy of Sciences, Dubravska cesta 21, 845 51, Bratislava, Slovak Republic.
Arch Microbiol. 2006 Dec;186(6):435-46. doi: 10.1007/s00203-006-0158-9. Epub 2006 Aug 15.
By using the previously established Escherichia coli two-plasmid system, we identified a promoter recognized by the Streptomyces coelicolor A3(2) stress-response sigma factor sigmaH. The promoter directed expression of the sigJ gene encoding an extracytoplasmic function (ECF) sigma factor. S1-nuclease mapping using RNA prepared from E. coli containing the two-plasmid system, and S. coelicolor A3(2) from various developmental stages identified an identical transcription start point in both strains, corresponding to the sigJp promoter. The sigJp promoter was induced during sporulation of aerial hyphae. The level of the transcript from sigJp was dramatically reduced in a S. coelicolor A3(2) sigH mutant and unaffected in a sigF mutant. The S. coelicolor A3(2) core RNA polymerase, after complementation with sigmaH, was able to recognize the sigJp promoter in vitro. A sigJ mutation had no obvious effect on growth, stress response, differentiation, and production of antibiotics. The results suggested that the S. coelicolor A3(2) sigJ gene is under the control of stress-response sigmaH, thus indicating a cascade of sigma factors in Streptomyces stress response and development. Considering the expression of sigJ and its direct dependence upon developmentally-regulated sigmaH, we assume that sigmaJ may play a role in the later stages of development of S. coelicolor A3(2).
通过使用先前建立的大肠杆菌双质粒系统,我们鉴定出了一个由天蓝色链霉菌A3(2)应激反应σ因子σH识别的启动子。该启动子指导编码胞外功能(ECF)σ因子的sigJ基因的表达。使用从含有双质粒系统的大肠杆菌以及处于不同发育阶段的天蓝色链霉菌A3(2)制备的RNA进行S1核酸酶图谱分析,确定了两种菌株中相同的转录起始点,对应于sigJp启动子。sigJp启动子在气生菌丝形成孢子的过程中被诱导。在天蓝色链霉菌A3(2)的sigH突变体中,来自sigJp的转录本水平显著降低,而在sigF突变体中则不受影响。天蓝色链霉菌A3(2)的核心RNA聚合酶在与σH互补后,能够在体外识别sigJp启动子。sigJ突变对生长、应激反应、分化和抗生素生产没有明显影响。结果表明,天蓝色链霉菌A3(2)的sigJ基因受应激反应σH的控制,从而表明链霉菌应激反应和发育过程中存在σ因子级联反应。考虑到sigJ的表达及其直接依赖于发育调控的σH,我们推测σJ可能在天蓝色链霉菌A3(2)发育的后期阶段发挥作用。