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Significance Cyclic di-GMP is a bacterial second messenger that transmits extracellular signals to the intracellular environment via sensor cyclic-di-GMP−metabolizing enzymes. Here a fluorescent biosensor is used to accurately measure cyclic-di-GMP concentrations in thousands of individual intracellularSalmonella Typhimurium. Furthermore, three enzymes that reduce cyclic-di-GMP concentrations were identified and shown to be essential for reduction of cyclic di-GMP, intracellular survival, and full virulence for mice. This was due to cyclic-di-GMP−mediated overproduction of cellulose that specifically affected a population of slowly replicating bacteria. These results further our knowledge of mechanisms of virulence and persistence of this important pathogen.

Cyclic-di-GMP regulation promotes survival of a slow-replicating subpopulation of intracellular Salmonella Typhimurium