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4. Pseudomonas aeruginosa zinc uptake in chelating environment is primarily mediated by the metallophore pseudopaline

Author

Brutesco, Catherine, Ghssein, Ghassan, Hajjar, Christine, Liratni, Ahmed, Wang, Shuanglong, Richaud, Pierre, Bleves, Sophie, Ball, Geneviève, Borezée-Durant, Elise, Lobinski, Ryszard, Pignol, David, Lhospice, Sébastien, Gomez, Nicolas Oswaldo, Ouerdane, Laurent, Arnoux, Pascal, and Voulhoux, Romé

Abstract

Metal uptake is vital for all living organisms. In metal scarce conditions a common bacterial strategy consists in the biosynthesis of metallophores, their export in the extracellular medium and the recovery of a metal-metallophore complex through dedicated membrane transporters. Staphylopine is a recently described metallophore distantly related to plant nicotianamine that contributes to the broad-spectrum metal uptake capabilities of Staphylococcus aureus. Here we characterize a four-gene operon (PA4837-PA4834) in Pseudomonas aeruginosa involved in the biosynthesis and trafficking of a staphylopine-like metallophore named pseudopaline. Pseudopaline differs from staphylopine with regard to the stereochemistry of its histidine moiety associated with an alpha ketoglutarate moiety instead of pyruvate. In vivo, the pseudopaline operon is regulated by zinc through the Zur repressor. The pseudopaline system is involved in nickel uptake in poor media, and, most importantly, in zinc uptake in metal scarce conditions mimicking a chelating environment, thus reconciling the regulation of the cnt operon by zinc with its function as the main zinc importer under these metal scarce conditions.

Published
2017

5. Visualization of the role of host heme on the virulence of the heme auxotroph Streptococcus agalactiae

Author

Joubert, Laétitia, Dagieu, Jean-Baptiste, Fernandez, Annabelle, Bobillot, Aurelie, Borezée-Durant, Elise, Fleurot, Isabelle, Gruss, Alexandra, Lechardeur, Delphine, MICrobiologie de l'ALImentation au Service de la Santé (MICALIS), Institut National de la Recherche Agronomique (INRA)-AgroParisTech, Fromageries Bel, Infectiologie Animale et Santé Publique - IASP (Nouzilly, France), Institut National de la Recherche Agronomique (INRA), HemeStockExchange ANR-12-BSV3-0022-01 project by the French 'Agence Nationale de la Recherche' - French National Research Agency ANR-11-IDEX-0003-02, 'ALIAS' project., Infectiologie et Santé Publique (UMR ISP), Institut National de la Recherche Agronomique (INRA)-Université de Tours (UT), ANR-12-BSV3-0022,HEMESTOCKEXCHANGE,Hème et porphyrine chez des bactéries modèle: des fonctions aux mécanismes(2012), ANR-11-IDEX-0003,IPS,Idex Paris-Saclay(2011), and Institut National de la Recherche Agronomique (INRA)-Université de Tours

Subjects
auxotrophie, Virulence, infection bactérienne, Intracellular Space, auxotrophy, toxicity, virulence factor, Aerobiosis, Article, Mice, ferroprotoporphyrin, facteur de virulence, [SDV.MP]Life Sciences [q-bio]/Microbiology and Parasitology, Genes, Bacterial, Streptococcal Infections, Animals, toxicité, streptococcus agalactiae, heme
Abstract

International audience; Heme is essential for several cellular key functions but is also toxic. Whereas most bacterial pathogens utilize heme as a metabolic cofactor and iron source, the impact of host heme during bacterial infection remains elusive. The opportunist pathogen Streptococcus agalactiae does not synthesize heme but still uses it to activate a respiration metabolism. Concomitantly, heme toxicity is mainly controlled by the HrtBA efflux transporter. Here we investigate how S. agalactiae manages heme toxicity versus benefits in the living host. Using bioluminescent bacteria and heme-responsive reporters for in vivo imaging, we show that the capacity of S. agalactiae to overcome heme toxicity is required for successful infection, particularly in blood-rich organs. Host heme is simultaneously required, as visualized by a generalized infection defect of a respiration-negative mutant. In S. agalactiae, HrtBA expression responds to an intracellular heme signal via activation of the two-component system HssRS. A hssRS promoter-driven intracellular luminescent heme sensor was designed to identify host compartments that supply S. agalactiae with heme. S. agalactiae acquires heme in heart, kidneys, and liver, but not in the brain. We conclude that S. agalactiae response to heme is organ-dependent, and its efflux may be particularly relevant in late stages of infection.

Published
2017

6. Pseudomonas aeruginosa zinc uptake in chelating environment is primarily mediated by the metallophore pseudopaline

Author

Lhospice, Sebastien, Gomez, Nicolas, Ouerdane, Laurent, Brutesco, Catherine, Ghssein, Ghassan, Hajjar, Christine, Liratni, Ahmed, Wang, Shuanglong, Richaud, Pierre, Bleves, Sophie, Ball, Geneviève, Borezée-Durant, Elise, Lobinski, Ryszard, Pignol, David, Arnoux, Pascal, Voulhoux, Romé, Laboratoire d'ingénierie des systèmes macromoléculaires (LISM), Aix Marseille Université (AMU)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Laboratoire d'Ingéniérie et Science des Matériaux - EA 4695 (LISM), Université de Reims Champagne-Ardenne (URCA)-SFR Condorcet, Université de Reims Champagne-Ardenne (URCA)-Centre National de la Recherche Scientifique (CNRS)-Université de Reims Champagne-Ardenne (URCA)-Centre National de la Recherche Scientifique (CNRS), Institut des sciences analytiques et de physico-chimie pour l'environnement et les materiaux (IPREM), Université de Pau et des Pays de l'Adour (UPPA)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Institut de Biosciences et Biotechnologies d'Aix-Marseille (ex-IBEB) (BIAM), Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS)-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Microbiologie Environnementale et Moléculaire (MEM), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS)-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Biologie végétale et microbiologie environnementale - UMR7265 (BVME), Environnement, Bioénergie, Microalgues et Plantes (EBMP), MICrobiologie de l'ALImentation au Service de la Santé (MICALIS), Institut National de la Recherche Agronomique (INRA)-AgroParisTech, Vaincre la Mucoviscidose [RFI20160501495], [ANR-14-CE09-0007-03], ANR-14-CE09-0007,ANIBAL,Un nouveau metallophore dérivé de la nicotianamine chez des bactéries pathogènes(2014), Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Aix Marseille Université (AMU), Université de Reims Champagne-Ardenne (URCA)-Université de Picardie Jules Verne (UPJV)-Centre National de la Recherche Scientifique (CNRS)-Université de Reims Champagne-Ardenne (URCA)-Université de Picardie Jules Verne (UPJV)-Centre National de la Recherche Scientifique (CNRS), Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS), Bioénergie et Microalgues (EBM), Université de Pau et des Pays de l'Adour (UPPA)-Centre National de la Recherche Scientifique (CNRS), Centre National de la Recherche Scientifique (CNRS)-Aix Marseille Université (AMU)-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), and Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre National de la Recherche Scientifique (CNRS)-Aix Marseille Université (AMU)-Direction de Recherche Fondamentale (CEA) (DRF (CEA))

Subjects
Zinc, Bacterial Proteins, lcsh:R, Operon, Pseudomonas aeruginosa, lcsh:Medicine, [CHIM]Chemical Sciences, lcsh:Q, Pseudomonas Infections, lcsh:Science, Oligopeptides, Article, Chelating Agents
Abstract

cited By 0; International audience; Metal uptake is vital for all living organisms. In metal scarce conditions a common bacterial strategy consists in the biosynthesis of metallophores, their export in the extracellular medium and the recovery of a metal-metallophore complex through dedicated membrane transporters. Staphylopine is a recently described metallophore distantly related to plant nicotianamine that contributes to the broad-spectrum metal uptake capabilities of Staphylococcus aureus. Here we characterize a four-gene operon (PA4837-PA4834) in Pseudomonas aeruginosa involved in the biosynthesis and trafficking of a staphylopine-like metallophore named pseudopaline. Pseudopaline differs from staphylopine with regard to the stereochemistry of its histidine moiety associated with an alpha ketoglutarate moiety instead of pyruvate. In vivo, the pseudopaline operon is regulated by zinc through the Zur repressor. The pseudopaline system is involved in nickel uptake in poor media, and, most importantly, in zinc uptake in metal scarce conditions mimicking a chelating environment, thus reconciling the regulation of the cnt operon by zinc with its function as the main zinc importer under these metal scarce conditions. © 2017 The Author(s).

Published
2017

15. Pseudomonas aeruginosa zinc uptake in chelating environment is primarily mediated by the metallophore pseudopaline.

Author

Lhospice S, Gomez NO, Ouerdane L, Brutesco C, Ghssein G, Hajjar C, Liratni A, Wang S, Richaud P, Bleves S, Ball G, Borezée-Durant E, Lobinski R, Pignol D, Arnoux P, and Voulhoux R

Subjects
Bacterial Proteins genetics, Pseudomonas aeruginosa genetics, Pseudomonas aeruginosa growth & development, Bacterial Proteins metabolism, Chelating Agents metabolism, Oligopeptides metabolism, Operon, Pseudomonas Infections microbiology, Pseudomonas aeruginosa metabolism, Zinc metabolism
Abstract

Metal uptake is vital for all living organisms. In metal scarce conditions a common bacterial strategy consists in the biosynthesis of metallophores, their export in the extracellular medium and the recovery of a metal-metallophore complex through dedicated membrane transporters. Staphylopine is a recently described metallophore distantly related to plant nicotianamine that contributes to the broad-spectrum metal uptake capabilities of Staphylococcus aureus. Here we characterize a four-gene operon (PA4837-PA4834) in Pseudomonas aeruginosa involved in the biosynthesis and trafficking of a staphylopine-like metallophore named pseudopaline. Pseudopaline differs from staphylopine with regard to the stereochemistry of its histidine moiety associated with an alpha ketoglutarate moiety instead of pyruvate. In vivo, the pseudopaline operon is regulated by zinc through the Zur repressor. The pseudopaline system is involved in nickel uptake in poor media, and, most importantly, in zinc uptake in metal scarce conditions mimicking a chelating environment, thus reconciling the regulation of the cnt operon by zinc with its function as the main zinc importer under these metal scarce conditions.

Published
2017
Full Text
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16. Visualization of the role of host heme on the virulence of the heme auxotroph Streptococcus agalactiae.

Author

Joubert L, Dagieu JB, Fernandez A, Derré-Bobillot A, Borezée-Durant E, Fleurot I, Gruss A, and Lechardeur D

Subjects
Aerobiosis drug effects, Animals, Genes, Bacterial, Heme toxicity, Intracellular Space drug effects, Intracellular Space metabolism, Mice, Streptococcal Infections microbiology, Streptococcal Infections pathology, Streptococcus agalactiae drug effects, Streptococcus agalactiae metabolism, Virulence drug effects, Heme metabolism, Streptococcus agalactiae pathogenicity
Abstract

Heme is essential for several cellular key functions but is also toxic. Whereas most bacterial pathogens utilize heme as a metabolic cofactor and iron source, the impact of host heme during bacterial infection remains elusive. The opportunist pathogen Streptococcus agalactiae does not synthesize heme but still uses it to activate a respiration metabolism. Concomitantly, heme toxicity is mainly controlled by the HrtBA efflux transporter. Here we investigate how S. agalactiae manages heme toxicity versus benefits in the living host. Using bioluminescent bacteria and heme-responsive reporters for in vivo imaging, we show that the capacity of S. agalactiae to overcome heme toxicity is required for successful infection, particularly in blood-rich organs. Host heme is simultaneously required, as visualized by a generalized infection defect of a respiration-negative mutant. In S. agalactiae, HrtBA expression responds to an intracellular heme signal via activation of the two-component system HssRS. A hssRS promoter-driven intracellular luminescent heme sensor was designed to identify host compartments that supply S. agalactiae with heme. S. agalactiae acquires heme in heart, kidneys, and liver, but not in the brain. We conclude that S. agalactiae response to heme is organ-dependent, and its efflux may be particularly relevant in late stages of infection.

Published
2017
Full Text
View/download PDF

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