ERKs and p38 kinase phosphorylate p53 protein at serine 15 in response to UV radiation

Phospholylation of the p53 tumor suppressor protein is Likely to play an im portant role in regulating its activity. Serine 15 phosphorylation of p53 l eads to a stabilization of p53 by reducing its interaction with murine doub le minute 2, a negative regulatory partner, Recently, p53 was reported to b e activated and phosphorylated at serine 15 following UV radiation. However , the signaling pathway that mediates UV-induced phosphorylation is less we ll characterized. Here, we provide evidence that UVB-induced phosphorylatio n of p53 at serine 15 is mediated directly by ERKs and p38 kinase. We find that in a mouse JB6 epidermal cell line, ERKs and p38 kinase form a complex with p53 following UVB radiation. Inhibition of ERKs or p38 kinase activit y by the use of a dominant negative mutant of ERK2 or p38 kinase or their r espective specific inhibitor, PD98059 or SB202190, results in abrogation of UVB-induced phosphorylation of p53 at serine 15. Strikingly, incubation of UVB-activated ERKs or p38 kinase immunoprecipitated complex with exogenous p53 shows serine 15 phosphorylation of both exogenous and co-precipitated endogenous p53 protein, Additionally, active recombinant ERR1/2 and p38 kin ase but not JNKs are also able to phosphorylate p53 at serine 15 in vitro. Furthermore, pretreatment of cells with PD98059 or SB202190 blocks p58-depe ndent transcription activity but increases the level of p53 co-precipitated murine double minute. These results strongly suggest that both ERKs and p3 8 kinase have a direct role in UVB-induced phosphorylation of p53 at serine 15 in vivo.