Aerobic glycerol dissimilation via the Enterococcus faecalis DhaK pathway depends on NADH oxidase and a phosphotransfer reaction from PEP to DhaK via EIIADha

Sauvageot, Nicolas; Ladjouzi, Rabia; Benachour, Abdellah; Rincé, Alain; Deutscher, Josef; Hartke, Axel

doi:10.1099/mic.0.061663-0

Cited by 12 publications

(5 citation statements)

References 19 publications

(16 reference statements)

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“…An interesting substrate to study in the context of this work was glycerol, since the complex catabolism of this sugar alcohol has been recently intensively studied in E. faecalis (28,29). E. faecalis harbors two pathways of glycerol catabolism, the glycerol kinase (GlpK) pathway and the dihydroxyacetone kinase (DHAK) pathway.…”

Section: Resultsmentioning

confidence: 99%

Loss of Antibiotic Tolerance in Sod-Deficient Mutants Is Dependent on the Energy Source and Arginine Catabolism in Enterococci

et al. 2015

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Enterococci are naturally tolerant to typically bactericidal cell wall-active antibiotics, meaning that their growth is inhibited but they are not killed even when exposed to a high concentration of the drug. The molecular reasons for this extraordinary tolerance are still incompletely understood. Previous work showed that resistance to killing collapsed specifically in mutants affected in superoxide dismutase (Sod) activity, arguing that bactericidal antibiotic treatment led to induction of a superoxide burst. In the present work, we show that loss of antibiotic tolerance in ⌬sodA mutants of pathogenic enterococci is dependent on the energy source present during antibiotic treatment. Hexoses induce greater killing than the pentose ribose, and no killing was observed with glycerol as the energy source. These results point to glycolytic reactions as crucial for antibiotic-mediated killing of ⌬sodA mutants. A transposon mutant library was constructed in Enterococcus faecalis ⌬sodA mutants and screened for restored tolerance of vancomycin. Partially restored tolerance was observed in mutants with transposon integrations into intergenic regions upstream of regulators implicated in arginine catabolism. In these mutants, the arginine deiminase operon was highly upregulated. A model for the action of cell wall-active antibiotics in tolerant and nontolerant bacteria is proposed. IMPORTANCEAntibiotic tolerance is a serious clinical concern, since tolerant bacteria have considerably increased abilities to resist killing by bactericidal drugs. Using enterococci as models for highly antibiotic-tolerant pathogens, we showed that tolerance of these bacteria is linked to their superoxide dismutase (Sod), arguing that bactericidal antibiotics induce generation of reactive oxygen species inside cells. Wild-type strains are tolerant because they detoxify these deleterious molecules by the activity of Sod, whereas Sod-deficient strains are killed. This study showed that killing depends on the energy source present during treatment and that an increase in arginine catabolism partially restored tolerance of the Sod mutants. These results are used to propose a mode-of-action model of cell wall-active antibiotics in tolerant and nontolerant bacteria. Even though enterococci are considered low-virulence pathogens, treatment of enterococcal infections is often difficult and lengthy. International guidelines for treatment of infective enterococcal endocarditis recommend 4 to 6 weeks of administration of penicillin or ampicillin plus an aminoglycoside, with a risk of acute renal failure due to the nephrotoxicity of the latter antibiotics (1-3). Enterococci, which are lactic acid bacteria and part of the human commensal flora, are naturally highly tolerant to antibiotics considered to be bactericidal such as penicillins and glycopeptides (4, 5). The mechanisms that allow these organisms to escape the lethal action of those drugs are still incompletely understood. On the basis of previous studies, it was suggested that the tolerance...

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Section: Resultsmentioning

confidence: 99%

Loss of Antibiotic Tolerance in Sod-Deficient Mutants Is Dependent on the Energy Source and Arginine Catabolism in Enterococci

et al. 2015

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“…A ptsHI operon in these two organisms is located elsewhere on the genomes of the two bacilli. Glycerol metabolism via the enzymes encoded by the dha operon and the adjacent genes in the two bacilli thus resembles the glycerol utilization systems present in L. monocytogenes (64) and E. faecalis (65), which first oxidize intracellular glycerol to dihydroxyacetone, which is subsequently phosphorylated by the DhaKLM system.…”

Section: Proteins Containing a Pts Component Fused To The N Or C Termmentioning

confidence: 99%

The Bacterial Phosphoenolpyruvate:Carbohydrate Phosphotransferase System: Regulation by Protein Phosphorylation and Phosphorylation-Dependent Protein-Protein Interactions

Deutscher

Aké

Derkaoui

et al. 2014

Microbiol Mol Biol Rev

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341

374

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SUMMARY The bacterial phosphoenolpyruvate (PEP):carbohydrate phosphotransferase system (PTS) carries out both catalytic and regulatory functions. It catalyzes the transport and phosphorylation of a variety of sugars and sugar derivatives but also carries out numerous regulatory functions related to carbon, nitrogen, and phosphate metabolism, to chemotaxis, to potassium transport, and to the virulence of certain pathogens. For these different regulatory processes, the signal is provided by the phosphorylation state of the PTS components, which varies according to the availability of PTS substrates and the metabolic state of the cell. PEP acts as phosphoryl donor for enzyme I (EI), which, together with HPr and one of several EIIA and EIIB pairs, forms a phosphorylation cascade which allows phosphorylation of the cognate carbohydrate bound to the membrane-spanning EIIC. HPr of firmicutes and numerous proteobacteria is also phosphorylated in an ATP-dependent reaction catalyzed by the bifunctional HPr kinase/phosphorylase. PTS-mediated regulatory mechanisms are based either on direct phosphorylation of the target protein or on phosphorylation-dependent interactions. For regulation by PTS-mediated phosphorylation, the target proteins either acquired a PTS domain by fusing it to their N or C termini or integrated a specific, conserved PTS regulation domain (PRD) or, alternatively, developed their own specific sites for PTS-mediated phosphorylation. Protein-protein interactions can occur with either phosphorylated or unphosphorylated PTS components and can either stimulate or inhibit the function of the target proteins. This large variety of signal transduction mechanisms allows the PTS to regulate numerous proteins and to form a vast regulatory network responding to the phosphorylation state of various PTS components.

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“…While some intracellular bacteria, such as Listeria monocytogenes and Mycobacterium tuberculosis , may prefer glycerol and fatty acids as carbon sources, respectively ( 30 ), our observations in S. aureus infecting both HaCaT and THP-1 cells indicate significant activation of DhaK, a dihydroxyacetone kinase involved in glycerol metabolism ( 31 ) ( Fig. 3A and B ).…”

Section: Resultsmentioning

confidence: 71%

Deciphering Staphylococcus aureus– host dynamics using dual activity-based protein profiling of ATP-interacting proteins

Ahator,

Hegstad,

Lentz

et al. 2024

mSystems

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The utilization of ATP within cells plays a fundamental role in cellular processes that are essential for the regulation of host–pathogen dynamics and the subsequent immune response. This study focuses on ATP-binding proteins to dissect the complex interplay between Staphylococcus aureus and human cells, particularly macrophages (THP-1) and keratinocytes (HaCaT), during an intracellular infection. A snapshot of the various protein activity and function is provided using a desthiobiotin-ATP probe, which targets ATP-interacting proteins. In S. aureus , we observe enrichment in pathways required for nutrient acquisition, biosynthesis and metabolism of amino acids, and energy metabolism when located inside human cells. Additionally, the direct profiling of the protein activity revealed specific adaptations of S. aureus to the keratinocytes and macrophages. Mapping the differentially activated proteins to biochemical pathways in the human cells with intracellular bacteria revealed cell-type-specific adaptations to bacterial challenges where THP-1 cells prioritized immune defenses, autophagic cell death, and inflammation. In contrast, HaCaT cells emphasized barrier integrity and immune activation. We also observe bacterial modulation of host processes and metabolic shifts. These findings offer valuable insights into the dynamics of S. aureus –host cell interactions, shedding light on modulating host immune responses to S. aureus , which could involve developing immunomodulatory therapies. IMPORTANCE This study uses a chemoproteomic approach to target active ATP-interacting proteins and examines the dynamic proteomic interactions between Staphylococcus aureus and human cell lines THP-1 and HaCaT. It uncovers the distinct responses of macrophages and keratinocytes during bacterial infection. S. aureus demonstrated a tailored response to the intracellular environment of each cell type and adaptation during exposure to professional and non-professional phagocytes. It also highlights strategies employed by S. aureus to persist within host cells. This study offers significant insights into the human cell response to S. aureus infection, illuminating the complex proteomic shifts that underlie the defense mechanisms of macrophages and keratinocytes. Notably, the study underscores the nuanced interplay between the host’s metabolic reprogramming and immune strategy, suggesting potential therapeutic targets for enhancing host defense and inhibiting bacterial survival. The findings enhance our understanding of host–pathogen interactions and can inform the development of targeted therapies against S. aureus infections.

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Aerobic glycerol dissimilation via the Enterococcus faecalis DhaK pathway depends on NADH oxidase and a phosphotransfer reaction from PEP to DhaK via EIIADha

Cited by 12 publications

References 19 publications

Loss of Antibiotic Tolerance in Sod-Deficient Mutants Is Dependent on the Energy Source and Arginine Catabolism in Enterococci

Loss of Antibiotic Tolerance in Sod-Deficient Mutants Is Dependent on the Energy Source and Arginine Catabolism in Enterococci

The Bacterial Phosphoenolpyruvate:Carbohydrate Phosphotransferase System: Regulation by Protein Phosphorylation and Phosphorylation-Dependent Protein-Protein Interactions

Deciphering Staphylococcus aureus– host dynamics using dual activity-based protein profiling of ATP-interacting proteins

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