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Proteomic profile of Campylobacter jejuni 11168 during in vitro growth in the presence or absence of penicillin

ABSTRACT: The proteomic makeup of three biological replicates of cells incubated for 24 h in MHB (control) or three biological replicates of cells incubated for 24 h in MHB containing 100 x MIC of penicillin G was analysed. The control or penicillin-treated cultures were stained with RSG and sorted using FACS into cells that did not fluoresce, cells that fluoresced weakly (dim) or cells that fluoresced brightly. Here we present the raw mass spectrometric data of control dimly stained cells and penicillin-treated brightly stained cells.

INSTRUMENT(S): LTQ Orbitrap Velos

ORGANISM(S): Campylobacter Jejuni Subsp. Jejuni Nctc 11168-bn148

TISSUE(S): Bacterial Protein

DISEASE(S): Gastroenteritis

SUBMITTER: Andrea Kovacs-Simon

LAB HEAD: Richard Titball

PROVIDER: PXD021418 | Pride | 2020-10-20

REPOSITORIES: Pride

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Project description:Subinhibitory concentrations of the neuroleptic drug thioridazine (TDZ) are well-known to enhance the killing of methicillin-resistant S. aureus (MRSA) by Î²-lactam antibiotics, however, the mechanism underlying the synergy between TDZ and Î²-lactams is not fully understood. In the present study we have examined the effect of a subinhibitory concentration of TDZ on antimicrobial resistance, the global transcriptome, and the cell wall composition of MRSA USA300. We show that TDZ is able to sensitize the bacteria to several classes of antimicrobials targeting the late stages of peptidoglycan synthesis. Furthermore, our microarray analysis demonstrates that TDZ modulates the expression of genes encoding membrane and surface proteins, transporters, and enzymes involved in amino acid biosynthesis. Interestingly, resemblance between the transcriptional profile of TDZ treatment and the transcriptomic response of S. aureus to known inhibitors of cell wall synthesis suggests that TDZ disturbs peptidoglycan biosynthesis at a stage that precedes transpeptidation. In support of this notion, dramatic changes in the muropeptide profile of USA300 were observed following growth in the presence of TDZ, indicating that TDZ can interfere with the formation of the pentaglycine branches. Strikingly, the addition of glycine to the growth medium relieved the effect of TDZ on the muropeptide profile. Furthermore, exogenous glycine offered a modest protective effect against TDZ-induced Î²-lactam sensitivity. We propose that TDZ exposure leads to a shortage of intracellular amino acids, including glycine, which is required for the production of normal peptidoglycan precursors with pentaglycine branches, the correct substrate of S. aureus penicillin-binding proteins. Collectively, this work demonstrates that TDZ has a major impact on the cell wall biosynthesis pathway in S. aureus and provides new insights into how MRSA may be sensitized towards Î²-lactam antibiotics. Staphylococcus aureus USA300 was grown to early exponential phase and treated with TDZ (16 Âµg/ml) alone or in combination with DCX (0.125 Âµg/ml) for 30 min. Changes in global gene expression were analyzed using the untreated culture as control. Hybridizations were performed in triplicate using RNA isolated from independent cultures.

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Proteomic profile of Campylobacter jejuni 11168 during in vitro growth in the presence or absence of penicillin

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