The Sle1 Cell Wall Amidase Controls Daughter Cell Splitting, Cell Size, and β-Lactam Resistance in Community Acquired Methicillin Resistant Staphylococcus aureus USA300

Most clinically relevant methicillin resistant Staphylococcus aureus (MRSA) strains have become resistant to {beta}-lactams antibiotics through horizontal acquisition of the mecA gene encoding PBP2a, a peptidoglycan transpeptidase with low affinity for {beta}-lactams. The level of resistance conferred by mecA is, however, strain dependent and the mechanisms underlying this phenomenon remain poorly understood. We here show that {beta}-lactam resistance correlates to expression of the Sle1 cell wall amidase in the fast spreading and highly virulent community-acquired MRSA USA300 clone. Sle1 is a substrate of the ClpXP protease, and while the high Sle1 levels in cells lacking ClpXP activity confer {beta}-lactam hyper-resistance, USA300 cells lacking Sle1 are as sensitive to {beta}-lactams as cells lacking mecA. This finding prompted us to assess the cellular roles of Sle1 in more detail, and we demonstrate that high Sle1 levels accelerate the onset of daughter cells splitting and decrease cell size. Vice versa, oxacillin decreases the Sle1 level, and imposes a cell-separation defect that is antagonized by high Sle1 levels, suggesting that high Sle1 levels increase tolerance to oxacillin by promoting cell separation. In contrast, increased oxacillin sensitivity of sle1 cells appears linked to a synthetical lethal effect on septum synthesis. In conclusion, this study demonstrates that Sle1 is a key factor in resistance to {beta}-lactam antibiotics in the JE2 USA300 model strain, and that PBP2a is required for expression of Sle1 in JE2 cells exposed to oxacillin.\n\nImportanceThe bacterium Staphylococcus aureus is a major cause of human disease, and the global spread of S. aureus resistant to {beta}-lactam antibiotics (MRSA) has made treatment increasingly difficult. {beta}-lactams interfere with cross-linking of the bacterial cell wall, however, the killing mechanism of this important class of antibiotics is still not fully understood. Here we provide novel insight into this topic by showing that {beta}-lactam resistance is controlled by the Sle1 cell wall amidase in the fast spreading and highly virulent MRSA USA300 clone. We show that Sle1 high levels accelerate the onset of daughter cells splitting and decrease cell size. Vice versa, oxacillin decreases the Sle1 level, and imposes a cell-separation defect that is antagonized Sle1. The key finding that resistance to {beta}-lactams correlates positively to expression of Sle1 indicates that, in S. aureus, the detrimental effects of {beta}-lactam antibiotics are linked to inhibition of daughter cells splitting.