Disrupting Central Carbon Metabolism Increases β-Lactam Antibiotic Susceptibility in Vibrio cholerae

Keller, Megan; Han, Xiaowei; Dörr, Tobias

doi:10.1128/jb.00476-22

Cited by 4 publications

(4 citation statements)

References 55 publications

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“…Regulation of bacterial metabolism has been identified as an additional mechanism that supports antibiotic resistance in Vibrio and other species ( 83 - 85 ). Indeed, a recent work has demonstrated that NaCl negatively regulates proton motive force (PMF) in Vibrio alginolyticus , leading to a reduction in aminoglycoside uptake and a subsequent increase in antibiotic resistance at high salt ( 86 ).…”

Section: Resultsmentioning

confidence: 99%

Low salinity activates a virulence program in the generalist marine pathogen Photobacterium damselae subsp. damselae

et al. 2023

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Facultative marine bacterial pathogens sense environmental signals so that the expression of virulence factors is upregulated on entry into hosts and downregulated during the free-living lifestyle in the environment. In this study, we utilized transcriptome sequencing to compare the transcriptional profiles of Photobacterium damselae subsp. damselae , a generalist pathogen that causes disease in diverse marine animals and fatal infections in humans at NaCl concentrations that mimic the free-living lifestyle or host internal milieu, respectively. We here show that NaCl concentration constitutes a major regulatory signal that shapes the transcriptome and uncover 1,808 differentially expressed genes (888 upregulated and 920 downregulated in response to low-salt conditions). Growth at 3% NaCl, a salinity that mimics the free-living lifestyle, upregulated genes involved in energy production, nitrogen metabolism, transport of compatible solutes, utilization of trehalose and fructose, and carbohydrate and amino acid metabolism with strong upregulation of the arginine deiminase system (ADS). In addition, we observed a marked increase in resistance to antibiotics at 3% NaCl. On the contrary, the low salinity conditions (1% NaCl) that mimic those encountered in the host triggered a virulence gene expression profile that maximized the production of the type 2 secretion system (T2SS)-dependent cytotoxins damselysin, phobalysin P, and a putative PirAB-like toxin, observations that were corroborated by the analysis of the secretome. Low salinity also upregulated the expression of iron-acquisition systems, efflux pumps, and other functions related to stress response and virulence. The results of this study greatly expand our knowledge of the salinity-responsive adaptations of a generalist and versatile marine pathogen. IMPORTANCE Pathogenic Vibrionaceae species experience continuous shifts of NaCl concentration in their life cycles. However, the impact of salinity changes in gene regulation has been studied in a small number of Vibrio species. In this study, we analyzed the transcriptional response of Photobacterium damselae subsp. damselae ( Pdd ), a generalist and facultative pathogen, to changes in salinity, and demonstrate that growth at 1% NaCl in comparison to 3% NaCl triggers a virulence program of gene expression, with a major impact in the T2SS-dependent secretome. The decrease in NaCl concentration encountered by bacteria on entry into a host is proposed to constitute a regulatory signal that upregulates a genetic program involved in host invasion and tissue damage, nutrient scavenging (notably iron), and stress responses. This study will surely inspire new research on Pdd pathobiology, as well as on other important pathogens of the family Vibrionaceae and related taxa whose salinity regulons still await investigation.

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Section: Resultsmentioning

confidence: 99%

Low salinity activates a virulence program in the generalist marine pathogen Photobacterium damselae subsp. damselae

et al. 2023

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“…This process provides precursors for all the biosynthetic reactions that are required for cell survival [66,67]. This makes central carbon metabolism i.e., carbon uptake and carbon utilization, an attractive antimicrobial target because these processes are typically essential for microbial survival [68][69][70]. Innate immunity, which frequently deprives bacteria of iron, amino acids, and other essential nutrients, serves as an example of the efficacy of this strategy [71][72][73].…”

Section: Central Carbon Metabolismmentioning

confidence: 99%

Providing insight into the mechanism of action of Cationic Lipidated Oligomers (CLOs) using metabolomics

Hussein,

Mahboob,

Tait

et al. 2024

Preprint

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The increasing resistance of clinically relevant microbes against current commercially available antimicrobials underpins the urgent need for alternative and novel treatment strategies. Cationic lipidated oligomers (CLOs) are innovative alternatives to antimicrobial peptides, and have reported antimicrobial potential. An understanding of their antimicrobial mechanism of action is required to rationally design future treatment strategies for CLOs, either in monotherapy or synergistic combinations. In the present study, metabolomics was used to investigate the potential metabolic pathways involved in the mechanisms of antibacterial activity of one CLO, C12-o-(BG-D)-10, which we have previously shown to be effective against methicillin-resistantStaphylococcus aureus(MRSA) ATCC 43300. The metabolomes of MRSA ATCC 43300 at 1, 3 and 6 h following treatment with C12-o-(BG-D)-10 (48 µg/mL i.e., 3x MIC) were compared to those of the untreated controls.Our findings reveal that the studied CLO, C12-o-(BG-D)-10, disorganized the bacterial membrane as the first step towards its antimicrobial effect, as evidenced by marked perturbations in the bacterial membrane lipids and peptidoglycan biosynthesis observed at early time points i.e., 1, and 3 h. Central carbon metabolism, and biosynthesis of DNA, RNA, and arginine were also vigorously perturbed, mainly at early time points. Moreover, bacterial cells were under osmotic and oxidative stress across all time points, evident by perturbations of trehalose biosynthesis and pentose phosphate shunt. Overall, this metabolomics study has, for the first time, revealed that the antimicrobial action of C12-o-(BG-D)-10 may potentially stem from the dysregulation of multiple metabolic pathways.ImportanceAntimicrobial resistance poses a significant challenge to healthcare systems worldwide. Novel anti-infective therapeutics are urgently needed to combat drug-resistant microorganisms. Cationic lipidated oligomers (CLOs) show promise as new antibacterial agents against Gram-positive pathogens likeStaphylococcus aureus(MRSA). Understanding their molecular mechanism(s) of antimicrobial action may help design synergistic CLO treatments along with monotherapy. Here, we describe the first metabolomics study to investigate the killing mechanism(s) of CLOs against MRSA. The results of our study indicate that the CLO, C12-o-(BG-D)-10, had a notable impact on the biosynthesis and organization of the bacterial cell envelope. C12-o-(BG-D)-10 also inhibits arginine, histidine, central carbon metabolism, and trehalose production, adding to its antibacterial characteristics. This work illuminates the unique mechanism of action of C12-o-(BG-D)-10 and opens an avenue to design innovative antibacterial oligomers/polymers for future clinical applications.

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“…Next, we set out to evaluate cell shape and cell differentiation as previous studies have reported that dysregulated central carbon metabolism often leads to defects in cell morphology [38,39]. Moreover, for growth on chitinaceous sources, V. parahaemolyticus is known to exhibit elongated cell morphologies as a response to environmental cues [21].…”

Section: Hexr Is a Regulator For Biofilm Formation Cell Differentiati...mentioning

confidence: 99%

“…Microscopy was performed as previously described [38], with minor adjustments. Overnight cultures were inoculated into LB and MM9 at a starting 𝑂𝐷 600𝑛𝑚 of 0.025 for 16 hours at 37C, 200rpm.…”

Section: Microscopymentioning

confidence: 99%

Functional genomics of chitin degradation byVibrio parahaemolyticusreveals finely integrated metabolic contributions to support environmental fitness

Getz,

Robinson,

Thomas

2024

Preprint

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Vibriospecies are marine prokaryotes that inhabit diverse ecological niches, colonizing abiotic and biotic surfaces. These bacteria are vital players in the global carbon cycle, assimilating billions of tonnes of chitin for carbon (and nitrogen) metabolites. Many bacterial proteins involved in the process-including chitinases, sugar transporters, and modifying enzymes-have been well studied. However, the genetic functional interplay and key drivers ofVibriocompetitive survival in the presence of chitin as the dominant carbon source is not understood. To address this question, we carried out transposon sequencing (Tn-seq) to determine the genetic fitness ofVibrio parahaemolyticusmutants grown on chitin as a sole carbon source. Along with validating knownVibriogenes associated with chitin metabolism, our data newly identified vital roles for an unclassified OprD-like import porin and a HexR family transcriptional regulator. Furthermore, we functionally implicated HexR in regulating multiple physiological processes involved inV. parahaemolyticusenvironmental survival including carbon assimilation and cell growth, biofilm formation, and cell motility. Notably, our data revealed a strict requirement for HexR in filamentous cell morphology, a critical trait forV. parahaemolyticusenvironmental fitness. Therefore, a vital import porin and genomic regulation mediated by HexR supports multiple physiological processes forVibriochitinolytic growth and environmental fitness.

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Disrupting Central Carbon Metabolism Increases β-Lactam Antibiotic Susceptibility in Vibrio cholerae

Cited by 4 publications

References 55 publications

Low salinity activates a virulence program in the generalist marine pathogen Photobacterium damselae subsp. damselae

Low salinity activates a virulence program in the generalist marine pathogen Photobacterium damselae subsp. damselae

Providing insight into the mechanism of action of Cationic Lipidated Oligomers (CLOs) using metabolomics

Functional genomics of chitin degradation byVibrio parahaemolyticusreveals finely integrated metabolic contributions to support environmental fitness

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