Liu, YiweiGloag, Erin S.Hill, Preston J.Parsek, Matthew R.Wozniak, Daniel J.El-Naggar, Mohamed Y.2022-12-142022-12-142022-050021-9193PMC9112932http://hdl.handle.net/10919/112893Pseudomonas aeruginosa and Staphylococcus aureus are two common pathogens causing chronic infections in the lungs of people with cystic fibrosis (CF) and in wounds, suggesting that these two organisms coexist <i>in vivo</i>. However, P. aeruginosa utilizes various mechanisms to antagonize S. aureus when these organisms are grown together <i>in vitro</i>. Here, we suggest a novel role for Psl in antagonizing S. aureus growth. Psl is an exopolysaccharide that exists in both cell-associated and cell-free forms and is important for biofilm formation in P. aeruginosa. When grown in planktonic coculture with a P. aeruginosa <i>psl</i> mutant, S. aureus had increased survival compared to when it was grown with wild-type P. aeruginosa. We found that cell-free Psl was critical for the killing, as purified cell-free Psl was sufficient to kill S. aureus. Transmission electron microscopy of S. aureus treated with Psl revealed disrupted cell envelopes, suggesting that Psl causes S. aureus cell lysis. This was independent of known mechanisms used by P. aeruginosa to antagonize S. aureus. Cell-free Psl could also promote S. aureus killing during growth in <i>in vivo</i>-like conditions. We also found that Psl production in P. aeruginosa CF clinical isolates positively correlated with the ability to kill S. aureus. This could be a result of P. aeruginosa coevolution with S. aureus in CF lungs. In conclusion, this study defines a novel role for P. aeruginosa Psl in killing S. aureus, potentially impacting the coexistence of these two opportunistic pathogens <i>in vivo</i>. IMPORTANCE Pseudomonas aeruginosa and Staphylococcus aureus are two important opportunistic human pathogens commonly coisolated from clinical samples. However, P. aeruginosa can utilize various mechanisms to antagonize S. aureus <i>in vitro</i>. Here, we investigated the interactions between these two organisms and report a novel role for P. aeruginosa exopolysaccharide Psl in killing S. aureus. We found that cell-free Psl could kill S. aureus <i>in vitro</i>, possibly by inducing cell lysis. This was also observed in conditions reflective of <i>in vivo</i> scenarios. In accord with this, Psl production in P. aeruginosa clinical isolates positively correlated with their ability to kill S. aureus. Together, our data suggest a role for Psl in affecting the coexistence of P. aeruginosa and S. aureus <i>in vivo</i>.application/pdfenIn CopyrightPseudomonas aeruginosaPslStaphylococcus aureuscell lysiscystic fibrosisexopolysaccharidepolymicrobialwoundEmerging Infectious DiseasesRare DiseasesClinical ResearchLungInfectious Diseases2.2 Factors relating to the physical environment2 AetiologyInfectionCongenitalHumansBiofilmsPseudomonas aeruginosaStaphylococcus aureusPseudomonas InfectionsStaphylococcal InfectionsCystic FibrosisPolysaccharidesMicrobial InteractionsArticle - Refereed2022-12-14https://doi.org/10.1128/jb.00076-222045Gloag, Erin [0000-0001-8895-3444]354461191098-5530