Enz S, Braun V, Crosa J H
Department of Molecular Microbiology and Immunology, School of Medicine, Oregon Health Sciences University, Portland 97201-3098, USA.
Gene. 1995 Sep 22;163(1):13-8. doi: 10.1016/0378-1119(95)00380-o.
We have analyzed the molecular mechanism of regulation of the ferric dicitrate transport system in Escherichia coli (Ec), by studying the transcription of the regulatory and structural genes under various environmental conditions, and by determining the location of their transcriptional start points and promoter regions. We report here that the main species observed in Northern hybridization analyses were a 2.5-kb mRNA, encoded by the outer membrane protein receptor gene fecA, and a 1.5-kb mRNA encoded by a region including the fecIR genes. The synthesis of the 2.5-kb fecA mRNA is regulated by both citrate and iron. Furthermore, transcription of fecA is dependent on the presence of FecI. The promoter region for the fecA mRNA, a likely site of action for FecI, is not related to the consensus promoter region for sigma 70 RNA polymerase in Ec K-12. However, it shows greatest similarity with promoters of genes regulated by a new sub-family of sigma factors, i.e., the extracytoplasmic function (ECF) sigma factors, which are associated with the expression of genes involved in extracytoplasmic functions, suggesting that FecI may act as a specialized sigma factor. We also show that the fecB,C,D,E transport genes are linked in operon fashion to fecA. Since the levels of the fecB,C,D,E RNAs are extremely low, as compared to the level of fecA mRNA, it is likely that processing from the 3' end must occur and stop near the end of fecA where a hairpin structure is located.
我们通过研究各种环境条件下调控基因和结构基因的转录,并确定其转录起始点和启动子区域的位置,分析了大肠杆菌(Ec)中铁柠檬酸转运系统的分子调控机制。我们在此报告,Northern杂交分析中观察到的主要转录本是由外膜蛋白受体基因fecA编码的2.5 kb mRNA,以及由包括fecIR基因的区域编码的1.5 kb mRNA。2.5 kb fecA mRNA的合成受柠檬酸盐和铁的双重调控。此外,fecA的转录依赖于FecI的存在。fecA mRNA的启动子区域可能是FecI的作用位点,与Ec K-12中σ70 RNA聚合酶的共有启动子区域无关。然而,它与由一个新的σ因子亚家族(即胞外功能(ECF)σ因子)调控的基因启动子具有最大的相似性,这些σ因子与参与胞外功能的基因表达相关,这表明FecI可能作为一种特殊的σ因子发挥作用。我们还表明,fecB、C、D、E转运基因以操纵子形式与fecA相连。由于与fecA mRNA的水平相比,fecB、C、D、E RNA的水平极低,因此很可能必须从3'端进行加工,并在fecA末端附近的发夹结构处停止。