| Abstrakt: |
ABSTRACTIn Staphylococcus aureus, the low-molecular-weight thiol called bacillithiol (BSH), together with cognate S-transferases, is believed to be the counterpart to the glutathione system of other organisms. To explore the physiological role of BSH in S. aureus, we constructed mutants with the deletion of bshA(sa1291), which encodes the glycosyltransferase that catalyzes the first step of BSH biosynthesis, and fosB(sa2124), which encodes a BSH-S-transferase that confers fosfomycin resistance, in several S. aureusstrains, including clinical isolates. Mutation of fosBor bshAcaused a 16- to 60-fold reduction in fosfomycin resistance in these S. aureusstrains. High-pressure liquid chromatography analysis, which quantified thiol extracts, revealed some variability in the amounts of BSH present across S. aureusstrains. Deletion of fosBled to a decrease in BSH levels. The fosBand bshAmutants of strain COL and a USA300 isolate, upon further characterization, were found to be sensitive to H2O2and exhibited decreased NADPH levels compared with those in the isogenic parents. Microarray analyses of COL and the isogenic bshAmutant revealed increased expression of genes involved in staphyloxanthin synthesis in the bshAmutant relative to that in COL under thiol stress conditions. However, the bshAmutant of COL demonstrated decreased survival compared to that of the parent in human whole-blood survival assays; likewise, the naturally BSH-deficient strain SH1000 survived less well than its BSH-producing isogenic counterpart. Thus, the survival of S. aureusunder oxidative stress is facilitated by BSH, possibly via a FosB-mediated mechanism, independently of its capability to produce staphyloxanthin. |
