Role of the NaHCO(3) Transporter MpsABC in the NaHCO(3)–β-Lactam-Responsive Phenotype in Methicillin-Resistant Staphylococcus aureus

Methicillin-resistant Staphylococcus aureus (MRSA) infections are an increasing concern due to their intrinsic resistance to most standard-of-care β-lactam antibiotics. Recent studies of clinical isolates have documented a novel phenotype, termed NaHCO(3) responsiveness, in which a substantial proportion of MRSA strains exhibit enhanced susceptibility to β-lactams such as cefazolin and oxacillin in the presence of NaHCO(3). A bicarbonate transporter, MpsAB (membrane potential-generating system), was recently found in S. aureus, where it plays a role in concentrating NaHCO(3) for anaplerotic pathways. Here, we investigated the role of MpsAB in mediating the NaHCO(3) responsiveness phenotype. Radiolabeled NaH(14)CO(3) uptake profiling revealed significantly higher accumulation in NaHCO(3)-responsive vs nonresponsive MRSA strains when grown in ambient air. In contrast, under 5% CO(2) conditions, NaHCO(3)-responsive (but not nonresponsive) strains exhibited repressed uptake. Oxacillin MICs were measured in four prototype strains and their mpsABC deletion mutants in the presence of NaHCO(3) supplementation under 5% CO(2) conditions. NaHCO(3)-mediated reductions in oxacillin MICs were observed in the responsive parental strains but not in mpsABC deletion mutants. No significant impact on oxacillin MICs was observed in the nonresponsive strains under the same conditions. Transcriptional and translational studies were carried out using both quantitative reverse transcription-PCR (qRT-PCR) and mpsA-green fluorescent protein (GFP) fusion constructs; these investigations showed that mpsA expression and translation were significantly upregulated during mid-exponential-phase growth in oxacillin-NaHCO(3)-supplemented medium in responsive versus nonresponsive strains. Taken together, these data show that the NaHCO(3) transporter MpsABC is a key contributor to the NaHCO(3)–β-lactam responsiveness phenotype in MRSA. IMPORTANCE MRSA infections are increasingly difficult to treat, due in part to their resistance to most β-lactam antibiotics. A novel and relatively common phenotype, termed NaHCO(3) responsiveness, has been identified in which MRSA strains show increased susceptibility in vitro and in vivo to β-lactams in the presence of NaHCO(3). A recently described S. aureus NaHCO(3) transporter, MpsAB, is involved in intracellular NaHCO(3) concentration for anaplerotic pathways. We investigated the role of MpsAB in mediating the NaHCO(3) responsiveness phenotype in four prototype MRSA strains (two responsive and two nonresponsive). We demonstrated that MpsABC is an important contributor to the NaHCO(3)–β-lactam responsiveness phenotype. Our study adds to the growing body of well-defined characteristics of this novel phenotype, which could potentially translate to alternative targets for MRSA treatment using β-lactams.