MexEF-OprN multidrug efflux pump transporter negatively controls N-acyl-homoserine lactone accumulation in pseudomonas syringae pv. Tabaci 6605

Abstract

Our previous studies revealed that flagellar-motility-defective mutants such as ∆fliC of Pseudomonas syringae pv. tabaci 6605 (Pta6605) have remarkably reduced production of N-acyl-homoserine lactones (AHL), quorum-sensing molecules. To investigate the reason of loss of AHL production in ∆fliC mutant, we carried out transposon mutagenesis. Among approximately 14,000 transconjugants, we found 11 AHL production-recovered (APR) strains. In these APR strains, a transposon was inserted into either mexE or mexF, genes encoding for the multidrug efflux pump transporter MexEF-OprN, and mexT, a gene encoding a putative transcriptional activator for mexEF-oprN. These results suggest that MexEF-OprN is a negative regulator of AHL production. To confirm the negative effect of MexEF-OprN on AHL production, loss- and gain-of-function experiments for mexEF-oprN were carried out. The ∆fliCmexF and ∆fliCmexT double mutant strains recovered AHL production, whereas the mexT overexpressing strain abolished AHL production, although the psyI, a gene encoding AHL synthase, is transcribed as wild type. Introduction of a mexF or mexT mutation into another flagellar-motility- and AHL production-defective mutant strain, ∆motCD, also recovered the ability to produce AHL. Furthermore, introduction of the mexF mutation into other AHL production-defective mutant strains such as ∆gacA and ∆aefR also recovered AHL production but not to the ∆psyI mutant. These results indicate that MexEF-OprN is a decisive negative determinant of AHL production and accumulation.

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Acknowledgements

We thank the Leaf Tobacco Research Laboratory, Japan Tobacco Inc., and Dr. D. Studholme, University of Exeter, UK, for providing Pta6605 and genome sequence information of Pta6605, respectively. This work was supported in part by Grants-in-Aid for Scientific Researches, 15H04458 and 16K14861 from the Ministry of Education, Culture, Sports, Science and Technology of Japan.

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Correspondence to Yuki Ichinose.

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Nucleotide sequence accession numbers: AB676069 for mexT and LC217528 for mexEFoprN, respectively.

Communicated by S. Hohmann

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Sawada, T., Eguchi, M., Asaki, S. et al. MexEF-OprN multidrug efflux pump transporter negatively controls N-acyl-homoserine lactone accumulation in pseudomonas syringae pv. Tabaci 6605. Mol Genet Genomics 293, 907–917 (2018). https://doi.org/10.1007/s00438-018-1430-9

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Keywords

  • N-Acyl-homoserine lactone
  • Flagella motility
  • MexEF-OprN
  • Multidrug efflux pump transporter
  • Quorum sensing