The synthesis of 3'-dATP and its use as an inhibitor of ATP-dependent DNA synthesis in toluene-treated Escherichia coli

Abstract

A structural analogue of ATP, 3'-deoxyadenosine triphosphate (3'-dATP), has been synthesized from cordycepin (3'-deoxyadenosine), characterized, and determined to be an inhibitor of ATP-dependent DNA synthesis in Escherichia coli cells which have been reduced permeable to nucleoside triphosphates by treatment with toluene. The analogue is a competitive inhibitor of ATP and it inhibits replicative DNA synthesis 50% at concentrations of ca. 0.15 mM in the presence of 1.0 mM ATP and 4 x 10(8) cells/ml. The degree of inhibition of a given amount of 3'-dATP is inversely related to the cell concentration in the reaction mixture. The analogue interferes with some function of ATP which is continuously required during the course of the reaction and does not irreversibly inactivate the cells' DNA synthesis apparatus. 3'-Deoxyadenosine triphosphate may prove useful in elucidating the roles of ATP in DNA synthesis in more purified replicating systems.