The tumor suppressor p53 can trigger cell death independently of its transcriptional activity through subcellular translocation and activation of proapoptotic Bcl-2 family members. The regulation of such activity of endogenous p53 in response to stress remains largely unknown. Here we show that nuclear, activated FOXO3a could impair p53 transcriptional activity. However, activation of FOXO3a either on serum starvation or by expressing a constitutively active form of FOXO3a could induce p53-dependent apoptosis, even in cells bearing a transcriptionally inactive form of p53. Furthermore, FOXO3a could promote p53 cytoplasmic accumulation by increasing its association with nuclear exporting machinery. Our data also suggest that PUMA and Bax are required for p53-dependent apoptosis in manner that is independent of p53 transcriptional activity.