Identification of PP1α as a caspase-9 regulator in IL-2 deprivation-induced apoptosis

One of the mechanisms that regulate cell death is the reversible phosphorylation of proteins. ERK/MAPK phosphorylates caspase-9 at Thr(125), and this phosphorylation is crucial for caspase-9 inhibition. Until now, the phosphatase responsible for Thr(125) dephosphorylation has not been described. Here, we demonstrate that in IL-2-proliferating cells, phosphorylated serine/threonine phosphatase type 1 alpha (PP1 alpha) associates with phosphorylated caspase-9. IL-2 deprivation induces PP1 alpha dephosphorylation, which leads to its activation and, as a consequence, dephosphorylation and activation of caspase-9 and subsequent dissociation of both molecules. In cell-free systems supplemented with ATP caspase-9 activation is induced by addition of cytochrome c and we show that in this process PP1 alpha is indispensable for triggering caspase-9 as well as caspase-3 cleavage and activation. Moreover, PP1 alpha associates with caspase-9 in vitro and in vivo, suggesting that it is the phosphatase responsible for caspase-9 dephosphorylation and activation. Finally, we describe two novel phosphatase-binding sites different from the previously described PP1 alpha consensus motifs, and we demonstrate that these novel sites mediate the interaction of PP1 alpha with caspase-9.