The influence of bcl-2 activity on 2'-deoxyadenosine-induced apoptosis was investigated in 697 human pre-B leukemia cells stably transfected with expression plasmid pHeBo-BCL-2alpha (697/BCL2 cells). Apoptosis was induced by the 2'-deoxyadenosine analogue, 2-chloro-2'-deoxyadenosine (Cl-dA), with the concentration for apoptosis in one-half of the cells at 24 hours (LD(50)) being 10 microM for 697 cells and 120 microM for 697/Bcl 2 cells. There was a strong positive correlation between Cl-dATP levels and apoptotic index (coefficient of determination, r(2)=0.95; P=0.027). When 697 cell and 697/Bcl 2 cell lines were treated with 5 microM Cl-dA, Cl-dATP did not significantly accumulate in the latter. The Cl-dATP/dATP ratio was 0.03 in Cl-dA treated 697/Bcl 2 cells but nearly 6 in treated 697 cells. Bcl 2 overproduction also suppressed the accumulation of dAMP, dADP and dATP in cells exposed to 2'-deoxyadenosine in the presence of pentostatin to abrogate the pronounced inversion of ATP/dATP pools associated with 2'-deoxyadenosine exposure. These results suggest that one consequence of bcl-2 activity is suppression of 2'-deoxyadenosine phosphorylation and elevation in the apoptotic target cells. Relief from deoxyadenylate stress imbalances implies a novel upstream site of bcl-2 activity.