Scholten M, Janssen R, Bogaarts C, van Strien J, Tommassen J
Department of Molecular Cell Biology, Utrecht University, The Netherlands.
Mol Microbiol. 1995 Jan;15(2):247-54. doi: 10.1111/j.1365-2958.1995.tb02239.x.
Growth of Escherichia coli K-12 in low-phosphate conditions results in the induction of the synthesis of many proteins, including the outer membrane porin PhoE, alkaline phosphatase, and the Pst system for the transport of phosphate (P1). This response is controlled by a two-component regulatory system of which PhoB and PhoR are the response-regulator and the sensor/kinase, respectively. When Shigella flexneri was starved for P1, neither PhoE nor alkaline phosphatase was produced. However, induction of the synthesis of the PstS protein was observed, indicating that S. flexneri contains a functional PhoB/PhoR regulatory system. Consistent with this notion, the introduction of the E. coli phoA gene in S. flexneri resulted in the induction of alkaline phosphatase synthesis under phosphate limitation. However, introduction of phoE on a plasmid did not lead to the expression of PhoE protein, indicating that S. flexneri PhoB does not recognize the phoE promoter region. The phoB gene was cloned and sequenced and in the deduced amino acid sequence two deviations from that of E. coli PhoB were detected. Site-directed mutagenesis revealed that one of these deviations, i.e. Leu-172, which is Arg in E. coli PhoB, is responsible for the lack of expression of the PhoE protein in S. flexneri.
大肠杆菌K-12在低磷酸盐条件下生长会导致许多蛋白质合成的诱导,包括外膜孔蛋白PhoE、碱性磷酸酶以及用于磷酸盐(P1)转运的Pst系统。这种反应由一个双组分调节系统控制,其中PhoB和PhoR分别是反应调节因子和传感器/激酶。当福氏志贺菌缺乏P1时,PhoE和碱性磷酸酶均未产生。然而,观察到PstS蛋白合成的诱导,表明福氏志贺菌含有功能性的PhoB/PhoR调节系统。与此观点一致,在福氏志贺菌中引入大肠杆菌phoA基因导致在磷酸盐限制下碱性磷酸酶合成的诱导。然而,在质粒上引入phoE并未导致PhoE蛋白的表达,表明福氏志贺菌PhoB不能识别phoE启动子区域。phoB基因被克隆并测序,在推导的氨基酸序列中检测到与大肠杆菌PhoB的两个偏差。定点诱变表明,这些偏差之一,即大肠杆菌PhoB中的Arg在福氏志贺菌PhoB中为Leu-172,是福氏志贺菌中PhoE蛋白缺乏表达的原因。