Stability of DNA replicating activity in permeabilized mouse cells during preincubation

Abstract

When permeabilized cells, treated with detergent and made permeable to the nucleoside triphosphates, were preincubated briefly without nucleoside triphosphates, the activity of DNA replication was lost rapidly. This loss of DNA replicating activity was prevented when the mixture of nucleoside triphosphates (5 mM ATP and 0.1 mM each of dATP, dGTP, dCTP and TTP, the same concentrations contained in reaction mixture) was added to the permeabilized cells during the incubation. Each of deoxyribonuclesode triphosphates or ribonucleoside triphosphates, when added at 5 mM, was effective to varying degrees, but ATP was the most effective. These results suggests that there exists a process or factor(s) that requires ATP for DNA replication in mammalian cells, and that its decay during the preincubation could be prevented by ATP.