The ColRS two-component system regulates membrane functions and protects Pseudomonas putida against phenol

As reported, the two-component system CoIRS is involved in two completely different processes. It facilitates the root colonization ability of Pseudomonas fluorescens and is necessary for the Tn4652 transposition-dependent accumulation of phenol-utilizing mutants in Pseudomonas putida. To determine the role of the CoIRS system in P. putida, we searched for target genes of response regulator CoIR by use of a promoter library. Promoter screening was performed on phenol plates to mimic the conditions under which the effect of CoIR on transposition was detected. The library screen revealed the porin-encoding gene oprQ and the alginate biosynthesis gene algD occurring under negative control of CoIR. Binding of CoIR to the promoter regions of oprQ and algD in vitro confirmed its direct involvement in regulation of these genes. Additionally, the porin-encoding gene ompA(PP0773) and the type I pilus gene csuB were also identified in the promoter screen. However, it turned out that ompA(PP0773) and csuB were actually affected by phenol and that the influence of CoIR on these promoters was indirect. Namely, our results show that CoIR is involved in phenol tolerance of P. putida. Phenol MIC measurement demonstrated that a colR mutant strain did not tolerate elevated phenol concentrations. Our data suggest that increased phenol susceptibility is also the reason for inhibition of transposition of Tn4652 in phenol-starving colR mutant bacteria. Thus, the current study revealed the role of the CoIRS two-component system in regulation of membrane functionality, particularly in phenol tolerance of P. putida.