Ser-46 of p53 is phosphorylated in response to DNA-damage in vivo, and it plays a pivotal role for apoptotic signaling by p53 through regulating the transcriptional activation of genes involved in apoptosis. We found that the presence of the nitric oxide (NO) donor S-nitroso-N-acetyl-D,L-penicillamine (200-800 muM) during UV-irradiation of MCF-7 cells resulted in a significant reduction in the Ser-46 phosphorylation, compared to the UV-irradiated cells without NO. This reduction occurred independently of cyclic GMP generation and without affecting activities of p53 kinases such as the PI3K family, p38 MAPK, and HIPK2. The presence of NO was found to protect HCT116 human colon tumor cells containing wild-type p53 from UV-induced apoptosis, whereas no apparent inhibitory effect of NO on UV-induced apoptosis was observed in those deficient in p53. Our results suggest that NO-mediated protection of apoptosis is p53-dependent, occurring at least partly through NO-inhibition of phosphorylation of p53 on Ser-46. (C) 2003 Elsevier Inc. All rights reserved.