Nitric oxide nitrates tyrosine residues of tumor-suppressor p53 protein in MCF-7 cells

It has been reported that mammalian cells incubated with excess nitric oxide (NO) accumulate p53 protein but concomitantly this p53 loses its capacity for binding to its DNA consensus sequence. As nitration of tyrosine residues in various proteins has been shown to inhibit their functions, we examined whether NO nitrates tyrosine residues in p53 protein. MCF-7 cells expressing wild-type p53 were incubated with S-nitrosoglutathione for 4 h and cellular extracts were immunoprecipitated with an anti-p53 antibody. Western blot analyses of immunoprecipitates for p53 or for nitrotyrosine revealed low levels of nitrotyrosine in p53 from untreated cells. Incubation with 2 mM S-nitrosoglutathione induced a significant increase in the nitrotyrosine level in p53 protein compared to nontreated cells. These results suggest that excess NO produced in inflamed tissues could nitrate p53 protein, playing a role in carcinogenesis by impairing functions of this tumor-suppressor protein. (C) 2000 Academic Press.