mechanism

The p53 protein plays a key role in secur-ing the apoptotic response of chronic lymphocytic leukemia (CLL) cells to geno-toxic agents. Transcriptional induction of proapoptotic proteins including Puma are thought to mediate p53-dependent apo-ptosis. In contrast, recent studies have identified a novel nontranscriptional mechanism, involving direct binding of p53 to antiapoptotic proteins including Bcl-2 at the mitochondrial surface. Here we show that the major fraction of p53 induced in CLL cells by chlorambucil, fludarabine, or nutlin 3a was stably associated with mitochondria, where it binds to Bcl-2. The Puma protein, which was constitutively expressed in a p53-independent manner, was modestly up-regulated following p53 induction. Pifithrin , an inhibitor of p53-mediated transcription, blocked the up-regulation of Puma and also of p21CIP1. Surprisingly, pifithrin dramatically augmented apo-ptosis induction by p53-elevating agents and also accelerated the proapoptotic conformation change of the Bax protein. These data suggest that direct interaction of p53 with mitochondrial antiapoptotic proteins including Bcl-2 is the major route for apoptosis induction in CLL cells and that p53’s transcriptional targets include proteins that impede this nontranscrip-tional pathway. Therefore, strategies that block up-regulation of p53-mediated tran-scription may be of value in enhancing apoptosis induction of CLL cells by p53-elevating drugs. (Blood. 2008;112: 3827-3834