The ABC transporter family efflux pump PvdRT‐OpmQ of <i>Pseudomonas putida</i> KT2440: purification and initial characterization

Stein, Nicola Victoria; Eder, Michelle; Brameyer, Sophie; Schwenkert, Serena; Jung, Heinrich

doi:10.1002/1873-3468.14601

Cited by 2 publications

(5 citation statements)

References 40 publications

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“…In search of these other transport systems, genes encoding components of tripartite efflux systems [PP_0166 ( paxA ), PP_0804, PP_0906 ( mexV ), PP_1264 ( fusBCD ), PP_1516 ( mexJ ), PP_2064 ( mexH ), PP_2818 ( mexD ), PP_3302, PP_3426 ( mexF ), PP_3455 ( parX ), PP_3549 ( emrA ), PP_5173 ( triC )] or outer membrane porins [PP_1798, PP_4519, PP_4923 (all tolC -like), PP_1019 ( oprB-I ), PP_1273, PP_2069, PP_2558, PP_3427 ( oprN )] were individually deleted in the Δ pm strain ( 23 ). Furthermore, based on a previous report on the involvement of type 6 secretion systems (T6SSs) in pyoverdine secretion by Pseudomonas taiwanesis ( 26 ), we introduced the Δ pm mutations into a P. putida KT2440R-derived strain (ΔT6SS) lacking all three T6SSs ( 25 ).…”

Section: Resultsmentioning

confidence: 99%

“…9 ). Nevertheless, to unequivocally demonstrate that the ParXY system is able to contribute to the secretion of the siderophore pyoverdine, biochemical evidence of direct binding and transport by the system is required, as has been provided in initial experiments for the PvdRT-OpmQ system ( 23 ). Finally, the increased resistance of the Δ parX strain to chloramphenicol suggests functional interactions of ParXY with other RND efflux systems (e.g., TtgABC).…”

Section: Resultsmentioning

confidence: 99%

“…Several efflux systems are proposed to be involved in this process: PvdRT-OpmQ, MdtABC-OpmB ( 15 ), and ParXY-OMP. Initial biochemical evidence for an interaction of PvdRT-OpmQ with pyoverdine has recently been published ( 23 ).…”

Section: Resultsmentioning

confidence: 99%

“…In P. putida KT2440, previous studies have shown that in addition to PvdRT-OpmQ, the tripartite efflux system MdtABC-OpmB (RND type) is also involved in the secretion of the siderophore ( 15 ). First biochemical evidence for an interaction of the PvdRT-OpmQ system of P. putida KT2440 with pyoverdine has been recently presented, reconfirming the involvement of this system in siderophore transport ( 23 ). Inactivation of these transport systems leads to the inhibition of pyoverdine secretion and, thus, the growth of the bacterium in the presence of iron deficiency.…”

Section: Introductionmentioning

confidence: 89%

“…The growth rate was calculated from the exponential phase of the growth curve. For growth recovery experiments using purified pyoverdine, the siderophore was extracted following a modified protocol based on Meyer et al ( 43 ), as recently described in Stein et al ( 23 ). All recovery experiments were performed in CAA and KB medium.…”

Section: Methodsmentioning

confidence: 99%

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The RND efflux system ParXY affects siderophore secretion in Pseudomonas putida KT2440

Stein,

Eder,

Burr

et al. 2023

Microbiol Spectr

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Tripartite efflux systems transport antimicrobial agents, toxic metabolites, and siderophores from Gram-negative cells into the environment. For example, the main siderophore pyoverdine of the soil bacterium Pseudomonas putida KT2440 is secreted into the environment via the PvdRT-OpmQ and MdtABC-OpmB systems. Here, we looked for efflux systems that might be involved in the secretion of pyoverdine in addition to the latter two systems. Screening of different efflux systems revealed that parX (encoding the periplasmic adapter protein of the ParXY system) is of particular importance for bacterial growth under iron limitation. Further analysis showed that the deletion of parX impairs the production and secretion of pyoverdine, causing the observed growth effect. The effects were dependent on the presence of other tripartite efflux systems and the conditions of iron limitation. The results suggest that ParXY not only plays a role in antibiotic resistance, as shown previously, but also influences the secretion of siderophores in a network of overlapping activities of different tripartite efflux systems. IMPORTANCE Gram-negative bacteria from the Pseudomonas group are survivors in various environmental niches. For example, the bacteria secrete siderophores to capture ferric ions under deficiency conditions. Tripartite efflux systems are involved in the secretion of siderophores, which are also important for antibiotic resistance. For one of these efflux systems, the resistance-nodulation-cell division transporter ParXY from the model organism Pseudomonas putida KT2440, we show that it influences the secretion of the siderophore pyoverdine in addition to its already known involvement in antibiotic resistance. Phenotypically, its role in pyoverdine secretion is only apparent when other pyoverdine secretion systems are inactive. The results confirm that the different tripartite efflux systems have overlapping substrate specificities and can at least partially functionally substitute for each other, especially in important physiological activities such as supplying the cell with iron ions. This fact must be taken into account when developing specific inhibitors for tripartite efflux systems.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 89%

Section: Methodsmentioning

confidence: 99%

See 3 more Smart Citations

The RND efflux system ParXY affects siderophore secretion in Pseudomonas putida KT2440

Stein,

Eder,

Burr

et al. 2023

Microbiol Spectr

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Total substitution and partial modification of the set of non-ribosomal peptide synthetases clusters lead to pyoverdine diversity in the Pseudomonas fluorescens complex

Graña-Miraglia,

Geney Higuita,

Salazar

et al. 2024

Front. Microbiol.

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Pyoverdines are high affinity siderophores produced by most Pseudomonas with a wide role in microbial interspecies interactions. They are primarily composed of a conserved chromophore moiety, an acyl side chain and a peptide backbone which may be highly variable among strains. Upon ferric iron sequestration, pyoverdines are internalized through specialized receptors. The peptide precursor of pyoverdine, termed ferribactin, is synthesized by a set of non-ribosomal peptide synthetase (NRPS) enzymes and further modified by tailoring enzymes. While PvdL, the NRPS responsible for the synthesis of the peptide moiety that derives into the chromophore is conserved, the NRPSs for the peptide backbone are different across fluorescent Pseudomonas. Although the variation of pyoverdine is a widely recognized characteristic within the genus, the evolutionary events associated with the diversity and distribution of this trait remain mostly unknown. This study analyzed the NRPSs clusters for the biosynthesis of the peptide backbone of ferribactin in the genomes of a representative subset of strains of the Pseudomonas fluorescens complex. Bioinformatic analysis of the specificity of adenylation domains of the NRPSs allowed the prediction of 30 different pyoverdine variants. Phylogenetic reconstruction and mapping of the NRPS clusters pinpointed two different general levels of modifications. In the first level, a complete replacement of the set of NRPRs by horizontal transfer occurs. In the second level, the original set of NRPSs is modified through different mechanisms, including partial substitution of the NRPS genes by horizontal transfer, adenylation domain specificity change or NRPS accessory domain gain/loss.

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The ABC transporter family efflux pump PvdRT‐OpmQ of Pseudomonas putida KT2440: purification and initial characterization

Cited by 2 publications

References 40 publications

The RND efflux system ParXY affects siderophore secretion in Pseudomonas putida KT2440

The RND efflux system ParXY affects siderophore secretion in Pseudomonas putida KT2440

Total substitution and partial modification of the set of non-ribosomal peptide synthetases clusters lead to pyoverdine diversity in the Pseudomonas fluorescens complex

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