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Mechanosensitive channels are ubiquitous in bacteria and provide an essential mechanism to survive sudden exposure to a hypo-osmotic environment by the sensing and release of increased turgor pressure. No mechanosensitive channels have thus far been identified and characterized for the human-specific bacterial pathogenNeisseria gonorrhoeae . In this study, we identified and characterized theN. gonorrhoeae MscS-like mechanosensitive channel (Ng-MscS). Electrophysiological analyses by the patch clamp method showed that Ng-MscS is stretch activated and contains pressure-dependent gating properties. Further mutagenesis studies of critical residues forming the hydrophobic vapor lock showed that gain-of-function mutations in Ng-MscS inhibited bacterial growth. Subsequent analysis of the function of Ng-MscS inN. gonorrhoeae by osmotic down-shock assays revealed that the survival of Ng-mscS deletion mutants was significantly reduced compared with that of wild-type strains, while down-shock survival was restored upon the ectopic complementation ofmscS . Finally, to investigate whether Ng-MscS is important forN. gonorrhoeae during infections, competition assays were performed by using a murine vaginal tract infection model. Ng-mscS deletion mutants were outcompeted byN. gonorrhoeae wild-type strains for colonization and survival in this infection model, highlighting that Ng-MscS contributes toin vivo colonization and survival. Therefore, Ng-MscS might be a promising target for the future development of novel antimicrobials.

Identification and Characterization of the Neisseria gonorrhoeae MscS-Like Mechanosensitive Channel