SSR42 is a novel regulator of cytolytic activity in <i>Staphylococcus aureus</i>

SSR42 is the longest noncoding RNA in the <i>Staphylococcus aureus</i> cell and the second-most abundant transcript in the stationary-phase transcriptome, second only to RNAIII. It is highly conserved across strains and exhibits pronounced stability in stationary phase; however, the mechanism behind its regulatory role has yet to be fully elucidated. Herein, we used transcriptomic and proteomic approaches to probe the role of SSR42, revealing that it is a powerful, novel activator of the primary leukocidin LukAB. SSR42 is required for cytotoxicity toward, and escape from within, human neutrophils, and also mediates survival within human blood. We show that SSR42 wields this role via derepression by the peroxide repressor PerR in response to the presence of human neutrophils and governs <i>lukAB</i> induction in this niche. Importantly, this regulation is driven by direct RNA-RNA interaction, as we show binding of the 5' untranslated region (UTR) of the <i>lukAB</i> transcript with the 3' end of SSR42, which ultimately modulates transcript stability as well as translational activity. Finally, we demonstrate that this behavior is absolutely required for full virulence of <i>S. aureus</i> in murine models of both pneumonia and sepsis. Collectively, we present SSR42 as a pleiotropic regulatory RNA that acts as a nexus between environmental sensing and the regulation of pathogenesis, responding to environmental stimuli and host immune factors to bolster cytotoxic behavior and facilitate infection in <i>S. aureus</i>.IMPORTANCE<i>Staphylococcus aureus</i> is a master pathogen due to its formidable collection of virulence factors. These are tightly controlled by a diverse group of regulators that titrate their abundance to adapt to unique infectious niches. The role of regulatory RNAs in stress adaptation and pathogenesis is becoming increasingly more relevant in <i>S. aureus</i>. In this study, we provide the most comprehensive global analysis to date of just such a factor, SSR42. Specifically, we uncover that SSR42 is required for mediating cytotoxicity-one of the pillars of infection-in response to phagocytosis by human neutrophils. We find that SSR42 is induced by components of the host immune system and facilitates downstream activation of cytotoxic factors via RNA-RNA interactions. This illustrates that SSR42 forms a pivotal link between sensing the external environment and mediating resistance to oxidative stress while promoting virulence, solidifying it as a major global regulator in <i>S. aureus</i>.