AntR-mediated bidirectional activation of antA and antR, anthranilate degradative genes in Pseudomonas aeruginosa.

ABSTRACT

Bidirectional activation of transcription is a peculiar regulation mode of gene expression. In this study, we show that genes involved in the metabolism of anthranilate, a precursor of biosynthesis of tryptophan and Pseudomonas quinolone signal (PQS) are regulated by this bidirectional activation of transcription. Anthranilate is degraded by anthranilate dioxygenase complex encoded by antABC operon, and AntR, a LysR-type regulator encoded by antR activates the transcription of antABC operon in the presence of anthranilate. In P. aeruginosa, antABC and antR are divergently located and AntR binds to the intergenic region between antA and antR to activate the antABC transcription. In this study, we determined the transcriptional start site of the antA promoter (antA(p)) and AntR-responsive elements (AREs) in P. aeruginosa. The upstream deletion analysis of antA(p) and in vitro gel shift assay with purified AntR showed that there are two AREs at -194 to -148 and -88 to -47 regions. We also found that AntR activates antR promoter (antR(p)) in the opposite direction and both AREs are important in the bidirectional activation of antA(p) and antR(p). Two AREs have different binding affinities to AntR and the strength of transcriptional activation was dramatically asymmetric depending on the direction. We suggest that the different affinities of two AREs may explain the asymmetry of the bidirectional activation by AntR.