The Second Type VI Secretion System of Pseudomonas aeruginosa Strain PAO1 Is Regulated by Quorum Sensing and Fur and Modulates Internalization in Epithelial Cells

Abstract

Background: Three T6SSs are present in P. aeruginosa.H1-T6SS secretes bacteriolytic toxins.Results: H2-T6SS is regulated by quorum sensing and Fur and modulates internalization in epithelial cells through PI3K-Akt host pathway activation.Conclusion: H2-T6SS plays a role in virulence.Significance: In contrast to the anti-prokaryotic H1-T6SS, H2-T6SS targets human cells.Those T6SSs can carry out different functions important in establishing infection.The genome of Pseudomonas aeruginosa PAO1 contains three type VI secretion systems (T6SSs) called H1-, H2-, and H3-T6SS.The H1-T6SS secretes three identified toxins that target other bacteria, providing a fitness advantage for P. aeruginosa, and likely contributes to bacterial pathogenesis in chronic infections.However, no specific substrates or defined roles have been described for the two other systems.Here, we demonstrate that the expression of H2-T6SS genes of strain PAO1 is up-regulated during the transition from exponential to stationary phase growth and regulated by the Las and Rhl quorum sensing systems.In addition, we identify two putative Fur boxes in the promoter region and find that H2-T6SS transcription is negatively regulated by iron.We also show that the H2-T6SS system enhances bacterial uptake into HeLa cells (75% decrease in internalization with a H2-T6SS mutant) and into lung epithelial cells through a phosphatidylinositol 3-kinase-dependent pathway that induces Akt activation in the host cell (50% decrease in Akt phosphorylation).Finally, we show that H2-T6SS plays a role in P. aeruginosa virulence in the worm model.Thus, in contrast to H1-T6SS, H2-T6SS modulates interaction with eukary-otic host cells.Together, T6SS can carry out different functions that may be important in establishing chronic P. aeruginosa infections in the human host.