Expression and Regulation of the Plasmid-encoded Hemolysin Determinant of Escherichia Coli

As a first approach towards studying the regulation of hemolysin synthesis in Escherichia coli, we have fused lacZ into the four hly genes (hlyC, hlyA, hlyBa and hlyBb) using the Mud-1 (Mu::lacZ, Y, Apr) phage. The sites of insertion of Mud-1 within the various hly genes of the Hly plasmid pHly152 were determined by the hemolytic phenotype of the Hly- mutants (Hly-ex/Hly-in or Hly-ex/Hly+in) and by complementation of these Hly- mutants with recombinant plasmids carrying cloned hly genes. It was found that hlyC, hlyA and hlyBa are transcribed from a relatively weak promoter (hlypL) located in front of hlyC. The activity of beta-galactosidase is considerably lower when Mud-1 is integrated in hlyBa than when it is inserted in hlyC, suggesting a considerable decline in hly gene expression from hlyC to hlyBa. The DNA sequence upstream of the coding region of hlyC was found to promote galK gene expression when a fragment covering this region was inserted into the promoter-probe vector pKO-11. A putative promoter sequence, which could correspond to hlypL, was identified in this sequence. The hlyBb gene appears to be transcribed from a different promoter and the direction of transcription seems to be opposite to that of the hlyC, A, Ba operon. The strength of this promoter (hlypR), based on the level of beta-galactosidase activity of Mud-1 insertion mutants in hlyBb, is considerably higher than that of hlypL.(ABSTRACT TRUNCATED AT 250 WORDS)