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Mechanisms affecting fluoride tolerance in the environmental bacterium Pseudomonas putida

ABSTRACT: Fluoride is the anionic form of fluorine, the 13th most abundant element in Earth's crust, and it is toxic to many organisms above a threshold concentration. Environmental bacteria can withstand relatively high fluoride concentrations, but the only mechanism described so far is the CrcB-dependent efflux. CrcB-mediated export is the primary mechanism of F-tolerance in the model environmental bacterium Pseudomonas putida, yet spontaneous NaF-tolerant mutants arise in the absence of the CrcB transporter, showing that this is not the sole pathway of tolerance. We used whole-genome sequencing, proteomic, and transcriptomic analyses to identify mechanisms that affect fluoride tolerance in Pseudomonas putida.

INSTRUMENT(S):

ORGANISM(S): Pseudomonas Putida Kt2440

SUBMITTER: Lea Ets

LAB HEAD: Maia Kivisaar

PROVIDER: PXD068884 | Pride | 2026-03-30

REPOSITORIES: Pride

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			Action	DRS
	WT1.raw	Raw
	WT2.raw	Raw
	WT3.raw	Raw
	WTNaF1.raw	Raw
	WTNaF2.raw	Raw

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Project description:Fluoride (F–) is the anionic form of fluorine, the 13th most abundant element in Earth crust, and it is toxic to many organisms above a threshold concentration. Environmental bacteria can withstand relatively high F– concentrations, but the only mechanism described so far is the CrcB dependent efflux. CrcB mediated export is the primary mechanism of F– tolerance in the model environmental bacterium Pseudomonas putida, yet spontaneous NaF-tolerant mutants arise in the absence of the CrcB transporter, showing that this is not the sole pathway of tolerance. We performed a genome wide search for factors affecting F– tolerance in P. putida. Screening >141,000 transposon mutants, we identified PP_3125, a Cro/cI type transcription regulator, as a determinant of high F– tolerance in a strain lacking the CrcB transporter. Proteomic and transcriptomic analyses revealed genes controlled by PP_3125, among which the benzoate transporter BenE-I was linked to the tolerance phenotype. These results show that transporters can adopt moonlighting functions, an aspect unexplored in bacteria. More broadly, the findings reveal alternative fluoride tolerance mechanisms in P. putida and define targets for engineering stress resilient microbial strains for industrial applications.

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