Molecular analysis of the promoter region of the Escherichia coli K-12 phoE gene : Identification of an element, upstream from the promoter, required for efficient expression of phoE protein

Abstract

The phoE gene of Escherichia coli codes for an outer membrane pore protein whose expression is induced under phosphate limitation. The promoter of this gene contains a 17 base-pair fragment, designated a pho box, which is present also in other phosphatecontrolled promoters. The mRNA start site was determined and found to be located downstream from the pho box, such that this element is located in the −35 region of the phoE promoter. A set of promoter deletions was generated in vitro and analysis of these deletions revealed that sequences upstream from the pho box are required for the efficient expression of phoE. The required upstream region is located (in part) between positions −106 and −121 relative to the mRNA start site, and contains sequences homologous to a pho box and a correctly spaced Pribnow box, but in the reversed orientation relative to the regular −35 and −10 regions. A proper spacing between this upstream region and the −35 region appears to be important, since an oligonucleotide insertion in the intervening region interferes with phoE expression. By cloning the upstream region in a lacZ operon fusion vector, a weak phosphate limitation-inducible promoter activity could be detected.