1887

Abstract

Three putative alkaline phosphatase genes, , and , were identified in the genome of by homology with the amino acid sequence obtained from the PhoA protein of . PhoA and PhoC correspond to broad-spectrum alkaline phosphatases whereas PhoD is similar to a Ca-dependent phospholipase D of . The and genes were efficiently expressed in R5 medium under phosphate-limited conditions, as shown by studies using the reporter gene, whereas was poorly transcribed under the same conditions. Expression of was clearly PhoP-dependent since it was not transcribed in the Δ mutant and was strongly activated under low phosphate concentrations. Similarly, expression of was PhoP-dependent and highly sensitive to phosphate availability. By contrast, expression of was not PhoP-dependent. Electrophoretic mobility shift assays showed that PhoP binds to the and promoters, but not to that of . Footprinting studies with GST–PhoP revealed the presence of a PHO box (two direct 11 nt repeats) in the promoter and two PHO boxes in the promoter of . The transcription start points of the three promoters were identified by primer extension. The transcription start point of coincides with the G of its translation start codon, indicating that this gene is transcribed as a leaderless mRNA. The deduced −10 and −35 regions of (but not those of ) overlapped with the PHO boxes in this promoter, suggesting that an excess of PhoP interferes with binding of the RNA polymerase to this promoter. In summary, the three promoters showed clear differences in the modulation of their expression by PhoP.

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2007-10-01
2020-04-07
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