title: The ColRS two-component system regulates membrane functions and protects Pseudomonas putida against phenol.
abstract: As reported, the two-component system ColRS 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 ColRS system in P. putida, we searched for target genes of response regulator ColR by use of a promoter library. Promoter screening was performed on phenol plates to mimic the conditions under which the effect of ColR on transposition was detected. The library screen revealed the porin-encoding gene oprQ and the alginate biosynthesis gene algD occurring under negative control of ColR. Binding of ColR 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 ColR on these promoters was indirect. Namely, our results show that ColR 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 ColRS two-component system in regulation of membrane functionality, particularly in phenol tolerance of P. putida.
authors.affiliation: Estonian Biocentre and Institute of Molecular and Cell Biology, Tartu University, 51010 Tartu, Estonia.
authors.forename: Paula Ann
authors.initials: PA
authors.lastname: Kivistik
authors.forename: Marta
authors.initials: M
authors.lastname: Putrins
authors.forename: Külliki
authors.initials: K
authors.lastname: Püvi
authors.forename: Heili
authors.initials: H
authors.lastname: Ilves
authors.forename: Maia
authors.initials: M
authors.lastname: Kivisaar
authors.forename: Rita
authors.initials: R
authors.lastname: Hõrak
mesh_terms: D000464:Alginates; D001425:Bacterial Outer Membrane Proteins / Q000235:genetics; D001426:Bacterial Proteins / Q000235:genetics*; D002473:Cell Wall / Q000502:physiology; D004251:DNA Transposable Elements / Q000235:genetics; D004305:Dose-Response Relationship, Drug; D010861:Fimbriae, Bacterial / Q000235:genetics; D017353:Gene Deletion; D015964:Gene Expression Regulation, Bacterial; D020723:Glucuronic Acid / Q000096:biosynthesis / Q000235:genetics; D006603:Hexuronic Acids; D019800:Phenol / Q000494:pharmacology*; D018272:Porins / Q000235:genetics / Q000378:metabolism; D011401:Promoter Regions, Genetic; D016958:Pseudomonas putida / Q000187:drug effects / Q000235:genetics*
chemical_list: D000464:Alginates; D001425:Bacterial Outer Membrane Proteins; D001426:Bacterial Proteins; D004251:DNA Transposable Elements; D006603:Hexuronic Acids; D018272:Porins; C078167:OMPA outer membrane proteins; D019800:Phenol; D020723:Glucuronic Acid