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Plasmid manipulation of bacterial behaviour through translational regulatory crosstalk

ABSTRACT: pQBR103 is a naturally occurring plasmid known to alter the behaviour of its host, Pseudomonas fluorescens. This plasmid encodes a predicted translational regulator, rsmQ which is a homologue of known global regulators within Pseudomonas. Within this study we compared Pseudomonas fluorescens SBW25 plasmid free cells with cells carrying either WT pQBR103 or pQBR103 lacking rsmQ. Using comparative quantitative proteomics and RNAseq we showed that the loss of rsmQ leads to widespread proteomic changes in the absence of changes in mRNA abundance, suggesting RsmQ is a translational regulator of its host chromosome.

INSTRUMENT(S): Orbitrap Eclipse

ORGANISM(S): Pseudomonas Fluorescens (strain Sbw25) Bacteria

SUBMITTER: Gerhard Saalbach

LAB HEAD: Jacob G Malone

PROVIDER: PXD033640 | Pride | 2023-01-13

REPOSITORIES: Pride

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Project description:Indole is ubiquitously synthesized by plants and bacteria and functions as an inter species signaling molecule to modulate a wide variety of cellular activities. However, it is not clear how the indole signal is perceived and responded by plant growth promoting rhizobacteria (PGPR) at the rhizosphere. Here, we demonstrated that indole enhanced antibiotic tolerance of Pseudomonas fluorescens 2P24, a PGPR well known for its biocontrol capacity. By conducting quantitative proteomic analysis, we showed that indole influences the expression of multiple genes including the emhABC operon encoding the major multidrug efflux pump in P. fluorescens 2P24. The indole-induced antibiotic tolerance was not related to bacterial dormancy or slow growth, but depended on the emhABC operon and the divergently transcribed TetR-like regulator emhR. By binding to the semi-palindromic operator sequence, EmhR repressed the expression of emhABC. It was further revealed that indole bound to EmhR and weakened the interaction between EmhR and the operator. This is consistent with our finding that indole-induced expression of the EmhABC efflux pump is dependent on EmhR. Using homology modeling and molecular dynamics simulation, we found that indole binding resulted in significantly decreased distance between the two DNA-recognizing α3 helices within the EmhR dimer, which would possibly account for its compromised DNA binding capacity. EmhR was further shown to globally influence protein expressions, especially transporters and proteins involved in the denitrification pathway. This EmhR-dependent, indole-induced antibiotic tolerance is likely to be prevalent in the Pseudomonas species, as the EmhR homologue in Pseudomonas syringae was also shown to be responsible for the indole-induced antibiotic tolerance. Taken together, our results revealed an indole-sensing transcription factor EmhR responsible for indole-induced antibiotic tolerance in Pseudomonas species and have important implications on the general mechanism for the indole sensing and responses in rhizobacteria.

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Plasmid manipulation of bacterial behaviour through translational regulatory crosstalk

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