A unique SaeS allele overrides cell-density dependent expression of saeR and lukSF-PV in the ST30-SCCmecIV lineage of CA-MRSA

Abstract ST30 (CC30)-SCC mec IV (USA1100) is one of the most common community-acquired methicillin-resistant Staphylococcus aureus (CA-MRSA) lineages. ST30 isolates typically carry lukSF-PV genes encoding the Panton-Valentine leukocidin (PVL) and are responsible for outbreaks of invasive infections worldwide. In this study, twenty CC30 isolates were analyzed. All were very susceptible to non-β-lactam antimicrobials, 18/20 harbored the lukSF-PV genes, only 1/20 exhibited agr - rnaIII dysfunction, and the majority was not able to form biofilm on inert surfaces. Analysis of lukSF-PV temporal regulation revealed that opposite to other CA-MRSA isolates, these genes were more highly expressed in early log phase than in stationary phase. This inverted lukSF-PV temporal expression was associated with a similar pattern of saeRS expression in the ST30 isolates, namely high level expression in log phase and reduced expression in stationary phase. Reduced saeRS expression in stationary phase was associated with low expression levels of the sae regulators, agr and agr -upregulator sarA , which activate the stationary phase sae -P1 promoter and overexpression of agr -RNAIII restored the levels of saeR and lukSF-PV trancripts in stationary phase. Altered SaeRS activity in the ST30 isolates was attributed to amino acid substitutions (N227S, E268K and S351T) in the HTPase_c domain of SaeS (termed SaeS SKT ). Complementation of a USA300 saeS mutant with the saeS SKT and saeS alleles under the direction of the log phase sae -P3 promoter revealed that saeR and lukSF-PV transcription levels were more significantly activated by saeS SKT than saeS. In summary our data identify a unique saeS allele ( saeS SKT ) which appears to override cell-density dependent SaeR and PVL expression in ST30 CA-MRSA isolates. Further studies to determine the contribution of saeS SKT allele to the pathogenesis of infections caused by ST30 isolates are merited.