Härle C, Kim I, Angerer A, Braun V
Universität Tübingen, Germany.
EMBO J. 1995 Apr 3;14(7):1430-8. doi: 10.1002/j.1460-2075.1995.tb07129.x.
Transport of ferric citrate into cells of Escherichia coli K-12 involves two energy-coupled transport systems, one across the outer membrane and one across the cytoplasmic membrane. Previously, we have shown that ferric citrate does not have to enter the cytoplasm of E. coli K-12 to induce transcription of the fec ferric citrate transport genes. Here we demonstrate that ferric citrate uptake into the periplasmic space between the outer and the cytoplasmic membranes is not required for fec gene induction. Rather, FecA and the TonB, ExbB and ExbD proteins are involved in induction of the fec transport genes independent of their role in ferric citrate transport across the outer membrane. The uptake of ferric citrate into the periplasmic space of fecA and tonB mutants via diffusion through the porin channels did not induce transcription of fec transport genes. Point mutants in FecA displayed the constitutive expression of fec transport genes in the absence of ferric citrate but still required TonB, with the exception of one FecA mutant which showed a TonB-independent induction. The phenotype of the FecA mutants suggests a signal transduction mechanism across three compartments: the outer membrane, the periplasmic space and the cytoplasmic membrane. The signal is triggered upon the interaction of ferric citrate with FecA protein. It is postulated that FecA, TonB, ExbB and ExbD transfer the signal across the outer membrane, while the regulatory protein FecR transmits the signal across the cytoplasmic membrane to FecI in the cytoplasm. FecI serves as a sigma factor which facilitates binding of the RNA polymerase to the fec transport gene promoter upstream of fecA.(ABSTRACT TRUNCATED AT 250 WORDS)
柠檬酸铁转运到大肠杆菌K - 12细胞中涉及两个能量偶联转运系统,一个穿过外膜,另一个穿过细胞质膜。此前,我们已表明柠檬酸铁不必进入大肠杆菌K - 12的细胞质就能诱导fec柠檬酸铁转运基因的转录。在此我们证明,fec基因诱导并不需要柠檬酸铁摄取到外膜和细胞质膜之间的周质空间。相反,FecA以及TonB、ExbB和ExbD蛋白参与fec转运基因的诱导,这与其在柠檬酸铁跨外膜转运中的作用无关。通过孔蛋白通道扩散将柠檬酸铁摄取到fecA和tonB突变体的周质空间中并不会诱导fec转运基因的转录。FecA中的点突变体在没有柠檬酸铁的情况下表现出fec转运基因的组成型表达,但仍需要TonB,只有一个FecA突变体除外,它显示出不依赖TonB的诱导。FecA突变体的表型提示了一种跨越三个区室的信号转导机制:外膜、周质空间和细胞质膜。该信号在柠檬酸铁与FecA蛋白相互作用时触发。据推测,FecA、TonB、ExbB和ExbD将信号跨外膜传递,而调节蛋白FecR将信号跨细胞质膜传递给细胞质中的FecI。FecI作为一种sigma因子,促进RNA聚合酶与fecA上游的fec转运基因启动子结合。(摘要截短于250字)