New insights into the regulation of the pac gene from Escherichia coli W ATCC 11105.
The regulation of the pac gene encoding the penicillin G acylase of Escherichia coli W ATCC 11015 has been investigated by a molecular approach using lacZ as a reporter gene. This analysis revealed that a region of 170 bp located upstream of the pac structural gene contains the regulatory sequences that control its expression. The cAMP receptor protein is involved not only in the catabolite repression of penicillin G acylase production caused by glucose but also in the induction of pac gene expression by phenylacetic acid. Primer extension analyses have demonstrated that the transcription of the pac gene can be initiated from at least three different promoters. Although all these promoters are functional, their relative activity depends on the transcribed gene, the P1 and P3 promoters being more active in the presence of the pac gene, whereas the P2 promoter was stronger when the upstream region of the pac gene was fused to the lacZ reporter. A deletion of the region surrounding the -10 box of the P3 promoter produced a constitutive expression of the fused gene indicating that this sequence is required for phenylacetic acid induction and suggesting that the expression of the pac gene is regulated by a repression mechanism. This work reveals that the regulation of the pac gene is more complex than previously envisioned and provides new clues to investigate further this interesting regulatory system.
Roa
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Department of Molecular Microbiology, Consejo Superior de Investigaciones Científicas, Madrid, Spain.
García
J L
JL
D001483:Base Sequence; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics*; D015964:Gene Expression Regulation, Bacterial; D005798:Genes, Bacterial; D017930:Genes, Reporter; D007700:Kinetics; D008969:Molecular Sequence Data; D010399:Penicillin Amidase / Q000235:genetics* / Q000378:metabolism*; D010957:Plasmids; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D001616:beta-Galactosidase / Q000235:genetics / Q000378:metabolism
D011993:Recombinant Fusion Proteins; D001616:beta-Galactosidase; D010399:Penicillin Amidase