Growth-phase-dependent expression of the Pseudomonas putida TOL plasmid pWW0 catabolic genes

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Abstract
Pseudomonas putida TOL plasmid pWW0 catabolic genes are clustered into two operons. The first, the upper operon, is controlled by the xylR regulatory gene, whereas the second, the meta operon, is controlled by the xylS regulatory gene. The xylS gene itself is subjected to control by xylR. In this study, we show that the TOL catabolic operons were poorly induced in cells growing at the early-exponential-growth phase but strongly induced in cells at late-exponential-growth phase. We constructed fusions of four TOL promoters, Pm (the promoter of the meta operon), Pu (the promoter of the upper operon), Ps (the promoter of the xylS regulatory gene), and Pr (the promoter of the xylR regulatory gene) with lacZ and examined, in Escherichia coli and P. putida, the expression of these promoters in relation to the growth phase. Expression from Pm, Pu, Ps, and Pr was almost constant if the host cells did not carry either xylS or xylR. Similarly, expression of Pm and Pu in P. putida in the absence of XylS and XylR was constant during the growth of the cells. XylS-dependent transcription of Pm and XylR-dependent transcription of Ps and Pu, in contrast, varied with the growth phase. This observation suggested that the interaction of XylS and XylR with target promoters or with RNA polymerases was influenced by the growth phase. The nature of the signal which triggers the growth-phase-dependent regulation was not clear. A change in the oxygen partial pressure was not responsible for the regulation. E. coli mutants defective in relA, crp, and cya exhibited growth-phase-dependent expression of the TOL catabolic genes, indicating that cyclic AMP and relA-dependent synthesis of ppGpp are not involved in this phenomenon.