Cross talk between catabolic pathways in Pseudomonas putida: XylS-dependent and -independent activation of the TOL meta operon requires the same cis-acting sequences within the Pm promoter.

The Pm promoter of the meta cleavage operon in the TOL (toluene degradation) plasmid pWW0 of Pseudomonas putida becomes activated by the plasmid-encoded XylS regulator in the presence of benzoate and certain substituted analogs such as 3-methylbenzoate. In the absence of XylS, Pm was still responsive to unsubstituted benzoate but with induction kinetics and a range of transcriptional activity which differed substantially from those for the XylS-mediated activation. XylS-independent induction by benzoate did not occur in a rpoN genetic background. Pm was also silent while cells were actively growing in rich medium. However, XylS-dependent transcription and XylS-independent transcription were initiated at the same nucleotide, as determined with primer extension mapping. Furthermore, a series of deletions and mutations at the Pm promoter sequence showed the same overall pattern of responsiveness to benzoate with and without XylS, thus providing genetic evidence that the same promoter structure is recognized and activated by at least two different regulators. One of them is XylS, while the other, provided by the host bacterium, could be related to the chromosome-encoded benzoate degradation pathway.