Extracellular Sensors and Extracellular Alarmones, Which Permit Cross-Talk Between Organisms, Determine the Levels of Alkali Tolerance and Trigger Alkali-Induced Acid Sensitivity in Escherichia Coli‪.‬

ABSTRACT For several stress responses in Escherichia coli, switching on involves conversion by the stress of an extracellular stress sensor (an extracellular sensing component, ESC) to an extracellular induction component (EIC), the latter functioning as an alarmone and inducing the response. The aim of this study was to establish whether alkali tolerance induction at pH 9.0, alkali sensitisation induced at pH 5.5 and the acid sensitisation induced at pH 9.0 involve sensing of pH changes by ESCs. The techniques involved made use of studies with cell-free culture filtrates. With respect to the inducible responses under test, these filtrates were prepared either from induced or uninduced cultures and filtrates from uninduced cultures were also activated in vitro, by the pH stress, in the absence of bacteria. Tests were then made to examine whether EICs (known to be needed for all these systems) are formed by activation, at the appropriate pH values, of filtrates from pH 7.0-grown cultures (i.e. uninduced culture filtrates); appearance of an EIC on activation would indicate the presence in the uninduced culture filtrate of an ESC. The studies showed that all three systems use ESCs to detect pH changes. Tests involving attempted enzymic and physical inactivation of the ESCs, and attempted removal of the ESCs by dialysis, showed that the ESC involved in alkali sensitisation is a small very heat-resistant protein. Strikingly, protease only partially inactivated the ESCs needed for alkali tolerance induction and for acid sensitisation; each system may be complex, involving both protein and non-protein (RNA?) ESCs, although other explanations are possible. It was also established that appropriate killed cultures can induce all three responses when incubated with pH 7.0-grown living cultures. The occurrence of ESC/EIC pairs for these three responses has led to the evolution of early warning systems for each, the diffusibility of the EICs, and their interaction with non-producers, allowing them to act pheromonally, inducing sensitive organisms to stress tolerance, prior to exposure to stressor.