PARP1 inhibitors regulate PARP1 structure independent of DNA, reducing binding affinity for single strand breaks

Abstract

AbstractCancers caused by mutations to the DNA repair machinery may be treated by inhibitors that target Poly(ADP-ribose) Polymerase 1 (PARP1). PARP inhibitors are thought to cause toxicity by trapping PARP1 at single strand breaks, preventing single strand break repair, thus leading to accumulation of DNA damage and cancer cell death. Intriguingly though, different PARP inhibitors display similar cellular toxicities and catalytic inhibition despite having widely varying trapping potencies. To better understand this apparent contradiction and identify complementary mechanisms of action, we here visualize the effect of inhibitors on individual PARP1 and PARP2 molecules by atomic force microscopy (AFM). We find, surprisingly, that inhibitors cause significant PARP1 compaction and loss of molecular flexibility also in the absence of DNA. This compaction correlates with the trapping potency of the inhibitor; and could be functionally relevant by reducing the subsequent binding of pre-treated PARP1 to DNA. Such changes are less pronounced for PARP2, which shares a high sequence identity with the PARP1 catalytic domain but lacks the DNA binding domain present in PARP1. Our findings reveal an additional, DNA-independent mechanism of action for PARP inhibitors, where PARP inhibitors with strong trapping potencies target PARP1 in the absence of DNA, compact their conformation and thereby reduce its ability to bind to DNA.