Transcriptional control of the nah and sal hydrocarbon-degradation operons by the nahR gene product

Abstract

The positively regulated nah and sal operons of the NAH7 plasmid from Pseudomonas putida encode the enzymes for metabolism of naphthalene via salicylate. To study their coordinate regulation, a 6-kb DNA fragment containing the entire nahA gene (encoding naphthalene dioxygenase), the gene of the nah operon, was cloned into a RSF1010 plasmid derivative. Analysis of expression of nahA from the nah promoter in either Escherichia coli or Pseudomonas putida showed that a 1.6-kb DNA fragment from the nahR (nah operon regulatory locus) region was required in trans for (i) induction by salicylate; (ii) high-level expression of nahA, and (iii) complementation of nahR- mutants. Measurement of transcription in induced and uninduced P. putida showed that induction of the nah and sal operons occurred at the transcriptional level. The trans-acting positive regulatory gene, nahR, however, was constitutively transcribed.