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  • Epigallocatechin gallate induces upregulation of the two-component VraSR system by evoking a cell wall stress response in Staphylococcus aureus.

Epigallocatechin gallate induces upregulation of the two-component VraSR system by evoking a cell wall stress response in Staphylococcus aureus.

Applied and environmental microbiology (2012-09-04)
Oren Levinger, Tamar Bikels-Goshen, Elad Landau, Merav Fichman, Roni Shapira
RESUMO

We previously found that a short exposure of Staphylococcus aureus to subinhibitory (SI) doses of epigallocatechin gallate (EGCG) results in increased cell wall thickness, adaptation, and enhanced tolerance to cell-wall-targeted antibiotics. In this study, the response to EGCG of sigB and vraSR transcription factor mutants was characterized. We show that in contrast to the results observed for wild-type (WT) strains, an S. aureus 315 vraSR null mutant exposed to SI doses of EGCG did not exhibit increased tolerance to EGCG and oxacillin. A diminished increase in tolerance to ampicillin (from 16-fold to 4-fold) and no change in the magnitude of resistance to vancomycin were observed. Preexposure to EGCG enhanced the tolerance of wild-type and sigB null mutant cells to lysostaphin, but this enhancement was much weaker in the vraSR null mutant. Marked upregulation (about 60-fold) of vraR and upregulation of the peptidoglycan biosynthesis-associated genes murA, murF, and pbp2 (2-, 5-, and 6-fold, respectively) in response to SI doses of EGCG were determined by quantitative reverse transcription-PCR (qRT-PCR). EGCG also induced the promoter of sas016 (encoding a cell wall stress protein of unknown function which is not induced in vraSR null mutants) in a concentration-dependent manner, showing kinetics comparable to those of cell-wall-targeting antibiotics. Taken together, our results suggest that the two-component VraSR system is involved in modulating the cell response to SI doses of EGCG.

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Sigma-Aldrich
Lysostaphin from Staphylococcus staphylolyticus, lyophilized powder, Protein 50-70 % by biuret, ≥500 units/mg protein
Sigma-Aldrich
Oxacillin sodium salt, 815-950 μg/mg (Oxacillin)
Sigma-Aldrich
Lysostaphin from Staphylococcus simulans, recombinant, expressed in E. coli, lyophilized powder
Sigma-Aldrich
Lysostaphin from Staphylococcus staphylolyticus, BioUltra, ≥97% (SDS-PAGE), Protein 40-60 % by biuret, ≥2,000 units/mg protein
Sigma-Aldrich
Oxacillin sodium salt monohydrate
Supelco
Oxacillin sodium salt monohydrate, VETRANAL®, analytical standard
Sigma-Aldrich
Lysostaphin from Staphylococcus staphylolyticus, aseptically filled