LAUSR

PARP1 inhibitors (PARPis) are used clinically during cancer therapy and are thought to exert their cytotoxicity through PARP1 polymerase inhibition and PARP1‐DNA trapping. Here, we showed no significant correlation between PARP1‐DNA trapping and cytotoxicity induced by PARPis. We complemented PARP1‐knockout sublines with wild‐type PARP1 and 11 mutants with different point mutations that affect the polymerase activity. When examining the PARPi talazoparib, the induced cytotoxicity was highly significantly correlated with cellular PARP1 polymerase activity, but not with its PARP1‐DNA trapping or polymerase inhibition. Similarly, talazoparib's PARP1‐DNA trapping revealed significant correlation with the polymerase activity rather than its inhibition. Differently, however, when evaluating purified wild‐type and mutated PARP1, we identified an almost linear relationship between PARPis’ inhibiting PARP1 dissociation from DNA and their cytotoxicity in 17 cancer cell lines. In contrast, no significant correlation existed between PARP1 polymerase inhibition in the histone‐based systems and the cytotoxicity. After careful comparisons on different methods and detection targets, we conclude that the PARPi‐mediated increase in PARP1‐DNA binding by inhibiting autoPARylation of PARP1 on DNA rather than in PARP1‐DNA trapping is correlated with PARPi's cytotoxicity. Accordingly, we established a new PARPi screening model that more closely predicts cytotoxicity.