Dataset Information

The role of the Ser/Thr kinase PrkC of Clostridium difficile in cell wall homeostasis and antimicrobial resistance.

ABSTRACT: Clostridium difficile is the leading cause of antibiotic associated diarrhea in adults. During infection, the bacteria must rapidly respond and adapt to the host environment by being able to activate survival strategies. Ser/Thr kinases (STKs) are involved in signal transduction and regulate numerous physiological processes. PrkC of C. difficile is STK with two PASTA domains. We showed that this protein was membrane associated and localized at the septum. We observed that prkC deletion affects the cell morphology with an increased mean size, heterogeneity in length and the presence of abnormal septa. A ∆prkC mutant was able to sporulate and germinate but had a reduced motility and formed more biofilms than the wild-type strain. Moreover, a ∆prkC mutant showed an increased sensitivity to antimicrobial compounds targeting the envelope such as deoxycholate, a secondary bile salt, cephalosporins acting on peptidoglycan synthesis, cationic antimicrobial peptides and lysozyme. This increased susceptibility was not associated to differences in the structure of peptidoglycan or of polysaccharide II (PSII). A proteomic study showed that the majority of C. difficile proteins associated to the cell wall are less abundant in the ∆prkC mutant compared to the WT strain. Finally, the ∆prkC mutant presented a delay in gut establishment in a hamster model of infection while its virulence was not significantly affected.

INSTRUMENT(S): Q Exactive HF

ORGANISM(S): Clostridioides Difficile

TISSUE(S): Culture Condition:anaerobically-grown Cell, Prokaryotic Cell

SUBMITTER: Thibaut Douché

LAB HEAD: Bruno Dupuy

PROVIDER: PXD012241 | Pride | 2019-06-14

REPOSITORIES: Pride

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Publications

The Ser/Thr Kinase PrkC Participates in Cell Wall Homeostasis and Antimicrobial Resistance in Clostridium difficile.

Cuenot Elodie E Garcia-Garcia Transito T Douche Thibaut T Gorgette Olivier O Courtin Pascal P Denis-Quanquin Sandrine S Hoys Sandra S Tremblay Yannick D N YDN Matondo Mariette M Chapot-Chartier Marie-Pierre MP Janoir Claire C Dupuy Bruno B Candela Thomas T Martin-Verstraete Isabelle I

Infection and immunity 20190723 8

Clostridium difficile is the leading cause of antibiotic-associated diarrhea in adults. During infection, C. difficile must detect the host environment and induce an appropriate survival strategy. Signal transduction networks involving serine/threonine kinases (STKs) play key roles in adaptation, as they regulate numerous physiological processes. PrkC of C. difficile is an STK with two PASTA domains. We showed that PrkC is membrane associated and is found at the septum. We o ...[more]

PMID: 31085703

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Project description:Multivalent membrane disruptors are a relatively new antimicrobial scaffold that are difficult for bacteria to develop resistance to and can act on both gram-positive and gram-negative bacteria. Nuclear Magnetic Resonance (NMR) metabolomics is an important method for studying resistance development in bacteria since it is both a quantitative and qualitative method to study and identify phenotypes by changes in metabolic pathways. Determine the likely metabolic differences between antimicrobially challenged and unchallenged growth and wild type and antimicrobially mutated Bacillus cereus (B. cereus) samples by using NMR hydrophilic metabolomics. Proton (1H) NMR hydrophilic metabolite analysis was conducted using B. cereus wild type and B. cereus that was mutated with C16-DABCO and mannose functionalized poly(amidoamine) dendrimers (DABCOMD). Both the wild type and the mutated sample types were grown in low levels of DABCOMD (challenged samples) or without the addition of DABCOMD to the growth media (unchallenged samples) for sample collection at the mid log and stationary phases and for growth curve procurement. Hierarchical clustering of only the challenged sample type showed that both the stationary phase sample types (mutant and wild type) clustered together while the both the mid log phase sample types were distinct. Hierarchical clustering of the unchallenged samples showed complete separation of all sample types. There were statistically significant (p-value and fold change) changes in the concentrations of metabolites in both energy related pathways and peptidoglycan synthesis between all sample types, especially with mutants and especially the challenged sample types have more N-acetylglucosamine (as much as a 94.2-fold increase). The mid log phase sample types showed a larger difference between sample types than their stationary phase counter parts. The challenged and unchallenged mutant samples showed a larger difference between sample types in comparison to the differences between the challenged and unchallenged wild type sample types. There was a larger metabolite difference when comparing the challenged mutant samples to the challenged wild type samples than when comparing the unchallenged mutant samples to the unchallenged wild type samples. The metabolomic analysis of wild type and multivalent DABCOMD mutated B. cereus under both challenged and unchallenged conditions indicated that the mutants, especially the challenged mutants, are likely changing their peptidoglycan layer to protect themselves from the high positive charge on the membrane disrupting DABCOMD. This membrane fortification most likely led to the slow growth curve of the mutated and especially the challenged mutant samples. The association of these sample types with metabolites associated with energy expenditure is attributed to the increased energy required for these changes to occur as well as to the decreased diffusion of nutrients across the membrane.

Dataset Information

The role of the Ser/Thr kinase PrkC of Clostridium difficile in cell wall homeostasis and antimicrobial resistance.

Publications

The Ser/Thr Kinase PrkC Participates in Cell Wall Homeostasis and Antimicrobial Resistance in Clostridium difficile.

Similar Datasets

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