Dataset Information

The regulator PltZ senses exogenous2,4-DAPG and regulate a novel multidrug resistant pump in involved in Pseudomonas aeruginosaATCC27853

ABSTRACT: 2, 4-diacetylphloroglucinol (2,4-DAPG) is secondary metabolites produced by soil rhizophsere bacterium Pseudomonas fluorescens with ability to against bacteria,fungi, oomycetes and nematodes. Pseudomonas aeruginos is considered to be one cause of nosocomial infection due to its multi-drug resistance. We found aromatic polyketide antibiotic2,4-DAPG promotes the expression ofmultidrug efflux pumpsgenes pltIJK by blocking the interaction between pltI promoter andTetR family transcription factor PltZin P. aeruginosa; and our proteomics data estsblished thatPltZ is involved in diverse physiological process including metabolism of amino acids, biosynthesis of virulence factors, motility, ion transporting, carbohydrate and lipid transporting.We described asophisticated ATP-binding cassette(ABC)transportsystems regulation mechanismwhich affords insights into the role ofaromatic polyketide antibiotics.

INSTRUMENT(S): Orbitrap Fusion Lumos

ORGANISM(S): Pseudomonas Aeruginosa Lesb58

TISSUE(S): Cell Culture

SUBMITTER: yu xiaoquan

LAB HEAD: Yong-Xing He

PROVIDER: PXD016202 | Pride | 2020-01-17

REPOSITORIES: Pride

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Publications

The Regulator PltZ Regulates a Putative ABC Transporter System PltIJKNOP of Pseudomonas aeruginosa ATCC 27853 in Response to the Antimicrobial 2,4-Diacetylphloroglucinol.

Guo Ding-Ding DD Luo Li-Ming LM Ma Hai-Long HL Zhang Si-Ping SP Xu Hang H Zhang Honghua H Wang Yong Y Yuan Yongna Y Wang Zhen Z He Yong-Xing YX

Frontiers in microbiology 20200708

Pseudomonas aeruginosa is an opportunistic pathogen commonly infecting immunocompromised patients with diseases like cystic fibrosis (CF) and cancers and has high rates of recurrence and mortality. The treatment efficacy can be significantly worsened by the multidrug resistance (MDR) of P. aeruginosa, and there is increasing evidence showing that it is easy for this pathogen to develop MDR. Here, we identified a gene cluster, pltZ-pltIJKNOP, which was originally assumed to b ...[more]

PMID: 32733400

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Project description:Pseudomonas fluorescens is implicated in food spoilage especially under cold storage, causing the increase of food waste and the reduction of environmental (and nutritional) sustainability. In food industry, Pseudomonas biofilm on pipe surfaces and facilities creates a persistent source of contamination and increases resistance to common disinfection strategies. Even though gene expression and protein pattern changes under environmental stimuli have been suggested, cellular mechanisms involved in biofilm production are poorly investigated for this species. In the present work, the cheese blue pigmenting P. fluorescens ITEM 17298 produced increased biofilm biomass and motility at 15 °C in comparison to 30 °C. Proteomic analysis revealed major differences of proteins involved in biofilm formation. In particular, at 15 °C we found increased amount of the c-di-GMP regulator PleD involved in motility as well as AlgB and the carbon storage regulator directly involved biofilm formation. Carbon metabolism, protein and fatty acid synthesis were also stimulated under cold incubation. Moreover, for the first time, enzymes correlated to indigoidine synthesis (blue pigment) were identified in the same condition. The differences in expression of regulatory proteins could therefore be the major determinant of the biofilm formation ability of P. fluorescens under refrigerated condition. Genomic and proteomic results also revealed virulence factors mostly detected at 30 °C. In order to counteract P. fluorescens persistence, this study also proposed a control strategy for biofilm development. Thus, a sub-lethal concentration of bovine lactoferrin hydrolysate (HLF) was assayed at both temperatures. Treatment caused a significant reduction in biofilm biomass, also confirmed by changes in proteome pattern. At each condition proteins involved regulatory functions and transcription factors were downregulated after HLF-treatment. Among these, at 15 °C we found that PleD was absent in treated samples, whilst negative regulators of alginate biosynthesis were detected; likewise, indigoidine related proteins were repressed. The cyclic-di-GMP-binding biofilm dispersal mediator was instead detected in all treated samples.

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.

Dataset Information

The regulator PltZ senses exogenous2,4-DAPG and regulate a novel multidrug resistant pump in involved in Pseudomonas aeruginosaATCC27853

Publications

The Regulator PltZ Regulates a Putative ABC Transporter System PltIJKNOP of <i>Pseudomonas aeruginosa</i> ATCC 27853 in Response to the Antimicrobial 2,4-Diacetylphloroglucinol.

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