Overlooked signals: Highly stable quorum sensing molecule in phage lysates induces quorum sensing response
Phage-bacterial interaction studies routinely apply phage lysates at final concentrations of up to 10% of the culture. Consequently, bacterial metabolites such as quorum sensing (QS) signaling molecules are transferred along with the phage lysate to recipient bacterial cells. Here, we show that the Pseudomonas aeruginosa QS molecules 3OC12-HSL and C4-HSL are rapidly degraded in phage lysates. In contrast, the hydrophobic QS molecule PQS is remarkably stable, for at least one year, due to its binding within outer membrane vesicles. Strikingly, we find that PQS exceeds concentrations of 10 {micro}M in standard phage lysate preparations. We show that PQS carried over from phage lysates induces QS-controlled production of the virulence factor pyocyanin in P. aeruginosa. This PQS carryover does not oppose previous conclusions of phage infection-induced PQS production, as we show here, that this response is also triggered by PQS-free phage lysates. Since other bacterial species, including Paracoccus and Vibrio harveyi, also produce hydrophobic QS molecules that are bound within outer membrane vesicles, it is likely that phage lysates from these bacteria may similarly contain stable QS molecules. Collectively, we demonstrate that membrane-bound QS molecules may significantly confound QS-related physiological outcomes of phage-host interaction studies. This can be avoided by using QS synthase mutants for phage propagation or by purifying phage particles from lysates to eliminate QS molecule carryover.