Statin-induced lipid carrier stress reveals a conserved vulnerability in β-lactam-resistant Gram-positive bacteria.

Methicillin-resistant Staphylococcus aureus resists β-lactam antibiotics through the allosteric transpeptidase penicillin-binding protein 2a, which operates within staphyloxanthin-rich membrane microdomains. Statins restore susceptibility by disrupting these microdomains and impairing penicillin-binding protein 2a oligomerization, but the mechanisms enabling resistance to this resensitization remain unclear. Here we show, using evolution experiments in strains lacking a functional staphyloxanthin pathway, that mutations in gdpP, a regulator of cyclic di-adenosine monophosphate signaling, are the predominant route for restoring oxacillin resistance during membrane microdomain disruption. This adaptation is blocked by simvastatin, revealing a synthetic lethal interaction. Mechanistically, simvastatin inhibits the mevalonate pathway, depleting the essential lipid carrier undecaprenyl phosphate and exacerbating peptidoglycan precursor imbalance, an effect phenocopied by lipid carrier-targeting antibiotics such as bacitracin. Although compensatory mutations can restore resistance, they impose a fitness cost in vivo. Importantly, this vulnerability extends to Streptococcus pneumoniae, revealing a conserved strategy to overcome β-lactam resistance in Gram-positive pathogens.