The effect of caffeine on cell growth and metabolism of thymidine in Escherichia coli.

(1) The influence of caffeine on growth and on the metabolism of thymidine was investigated in various E. coli strains. Caffeine caused filamentous growth in all strains investigated. The caffeine effect was reversible. (2) The incorporation of thymidine into DNA was inhibited by caffeine, and the inhibition was most pronounced with bacterial cultures grown overnight in the presence of caffeine before the addition of thymidine. For cells not pretreated with caffeine the inhibitory effect of caffeine decreased with increasing concentrations of thymidine up to about 1 microM whereafter it remained constant. The effect of thymidine concentration on the inhibition was less for bacteria that had grown overnight in the presence of caffeine than for bacteria not pretreated with caffeine. (3) Caffeine inhibited thymidine kinase, but it had no effect on thymidine phosphorylase or thymidine nucleotide kinases. (4) It is suggested that caffeine interferes with uptake of thymidine, conversion of thymidine to dTTP and the DNA synthesis process itself. Filamentous growth could be the result of the inhibition of DNA synthesis.