Impact of a Transposon Insertion in
            <i>phzF2</i>
            on the Specialized Metabolite Production and Interkingdom Interactions of Pseudomonas aeruginosa

Phelan, Vanessa V.; Moree, Wilna J.; Aguilar, Julieta; Cornett, Dale S.; Koumoutsi, Alexandra; Noble, Suzanne M.; Pogliano, Kit; Guerrero, C.; Dorrestein, Pieter C.

doi:10.1128/jb.01258-13

Cited by 36 publications

(39 citation statements)

References 55 publications

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“…As shown in Figure 1a wide variety of specialized metabolites were detected including the phenazines pyocyanin (PYO), phenazine-1-carboxamide (PCN) and phenazine-1-carboxylic acid (PCA); quinolones 2-heptyl-4-quinolone (HHQ), PQS, 4-hydroxy-2-heptylquinoline- N -oxide (HQNO), 2-nonyl-4-quinolone (NHQ) and 2-nonenyl-hydroxy-4-quinolone (C9:db PQS) and 4-hydroxy-2-nonenylquinoline- N -oxide (C9:db NQNO) among others; various rhamnolipids as exemplified by 3-[3-[(2R,3R,4R,5R,6S)-4,5-dihydroxy-6-methyl-3-[(2S,3R,4R,5R,6S)-3,4,5-trihydroxy-6-methyloxan-2-yl]oxyoxan-2-yl]oxydecanoyloxy]decanoic acid (Rha-Rha-C10-C10) and the siderophores pyochelin and pyoverdine. [20, 21]…”

Section: Resultsmentioning

confidence: 99%

“…Structural verification of ions putatively identified in IMS was performed manually by comparing the exact mass from the LC-MS/MS data and the corresponding MS/MS spectra with previously reported structural characterization. [20, 21] LC-MS/MS data analysis was performed using Bruker Daltronics DataAnalysis v4.1 (Build 362.7). Lock mass internal calibration using hexakis(1H,1H,3H-tetrafluoropropoxy)phosphazene ions (Synquest Laboratories; m/z 922.0098) was applied.…”

Section: Methodsmentioning

confidence: 99%

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Mass Spectrometry Analysis of Pseudomonas aeruginosa Treated with Azithromycin

Phelan

Fang

Dorrestein

2015

J. Am. Soc. Mass Spectrom.

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In microbiology, changes in specialized metabolite production (cell-to-cell signaling metabolites, virulence factors and natural products) are measured using phenotypic assays. However, advances in mass spectrometry based techniques including imaging mass spectrometry (IMS) now allow researchers to directly visualize the production of specialized metabolites from microbial colony biofilms. In this study, a combination of IMS and liquid chromatography-tandem mass spectrometry (LC-MS/MS) was used to visualize the effect of the macrolide antibiotic azithromycin (AZM) on colony biofilms of Pseudomonas aeruginosa. While previous research suggested that AZM may inhibit cell-to-cell signaling of P. aeruginosa and thereby reducing pathogenicity, we observed no clear decrease in specialized metabolite production.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Mass Spectrometry Analysis of Pseudomonas aeruginosa Treated with Azithromycin

Phelan

Fang

Dorrestein

2015

J. Am. Soc. Mass Spectrom.

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“…The bacterial-fungal interplay is more complex than just molecules produced by Pa , as recent elegant work has shown that Af can transform Pa metabolites, and thus radically alter the effect on the interaction, including the degree of inhibition [ 65 ]. Finally, Fe limitation by Pa could also in part be a result of modulation of Af siderophore production by Pa metabolites [ 66 ].…”

Section: Discussionmentioning

confidence: 99%

Inhibition of Aspergillus fumigatus and Its Biofilm by Pseudomonas aeruginosa Is Dependent on the Source, Phenotype and Growth Conditions of the Bacterium

et al. 2015

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Aspergillus fumigatus (Af) and Pseudomonas aeruginosa (Pa) are leading fungal and bacterial pathogens, respectively, in many clinical situations. Relevant to this, their interface and co-existence has been studied. In some experiments in vitro, Pa products have been defined that are inhibitory to Af. In some clinical situations, both can be biofilm producers, and biofilm could alter their physiology and affect their interaction. That may be most relevant to airways in cystic fibrosis (CF), where both are often prominent residents.We have studied clinical Pa isolates from several sources for their effects on Af, including testing involving their biofilms. We show that the described inhibition of Af is related to the source and phenotype of the Pa isolate. Pa cells inhibited the growth and formation of Af biofilm from conidia, with CF isolates more inhibitory than non-CF isolates, and non-mucoid CF isolates most inhibitory. Inhibition did not require live Pa contact, as culture filtrates were also inhibitory, and again non-mucoid>mucoid CF>non-CF. Preformed Af biofilm was more resistant to Pa, and inhibition that occurred could be reproduced with filtrates. Inhibition of Af biofilm appears also dependent on bacterial growth conditions; filtrates from Pa grown as biofilm were more inhibitory than from Pa grown planktonically. The differences in Pa shown from these different sources are consistent with the extensive evolutionary Pa changes that have been described in association with chronic residence in CF airways, and may reflect adaptive changes to life in a polymicrobial environment.

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“…S10C). Chemically synthesized 5-Me-PCA and the 5-Me-PCA analog PMS are yellow (52), and PMS takes on a bright green color upon reduction (53). Accumulation of a reduced form of 5-Me-PCA could occur under the reducing conditions prevailing in the highcell density, oxygen-limited conditions of stationary phase cultures; if reduced 5-Me-PCA is green, this could account for the coloration of ΔphzHS cultures.…”

Section: Mexghi-opmd Contributes To Fitness In Shaken Liquid Culturesmentioning

confidence: 99%

The Pseudomonas aeruginosa efflux pump MexGHI-OpmD transports a natural phenazine that controls gene expression and biofilm development

Sakhtah

Koyama

Zhang

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

145

159

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Redox-cycling compounds, including endogenously produced phenazine antibiotics, induce expression of the efflux pump MexGHIOpmD in the opportunistic pathogen Pseudomonas aeruginosa. Previous studies of P. aeruginosa virulence, physiology, and biofilm development have focused on the blue phenazine pyocyanin and the yellow phenazine-1-carboxylic acid (PCA). In P. aeruginosa phenazine biosynthesis, conversion of PCA to pyocyanin is presumed to proceed through the intermediate 5-methylphenazine-1-carboxylate (5-Me-PCA), a reactive compound that has eluded detection in most laboratory samples. Here, we apply electrochemical methods to directly detect 5-Me-PCA and find that it is transported by MexGHIOpmD in P. aeruginosa strain PA14 planktonic and biofilm cells. We also show that 5-Me-PCA is sufficient to fully induce MexGHI-OpmD expression and that it is required for wild-type colony biofilm morphogenesis. These physiological effects are consistent with the high redox potential of 5-Me-PCA, which distinguishes it from other well-studied P. aeruginosa phenazines. Our observations highlight the importance of this compound, which was previously overlooked due to the challenges associated with its detection, in the context of P. aeruginosa gene expression and multicellular behavior. This study constitutes a unique demonstration of efflux-based selfresistance, controlled by a simple circuit, in a Gram-negative pathogen.phenazine | RND efflux | MexGHI-OpmD | biofilm | antibiotic

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Impact of a Transposon Insertion in phzF2 on the Specialized Metabolite Production and Interkingdom Interactions of Pseudomonas aeruginosa

Cited by 36 publications

References 55 publications

Mass Spectrometry Analysis of Pseudomonas aeruginosa Treated with Azithromycin

Mass Spectrometry Analysis of Pseudomonas aeruginosa Treated with Azithromycin

Inhibition of Aspergillus fumigatus and Its Biofilm by Pseudomonas aeruginosa Is Dependent on the Source, Phenotype and Growth Conditions of the Bacterium

The Pseudomonas aeruginosa efflux pump MexGHI-OpmD transports a natural phenazine that controls gene expression and biofilm development

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