The p53R2 ribonucleotide reductase subunit is a p53-inducible protein involved in DNA repair and mitochondrial DNA replication. It has been shown that p53 is activated by nitric oxide, which can damage DNA at high concentrations. This suggests that NO may regulate p53R2 expression through p53 activation. We show here that NO increases p53 protein expression in p53-wt cell lines and upregulates p53R2 at the protein and mRNA levels in a p53-dependent manner. Other p53 target genes, such as DDB2, WAF1 and PCNA, are also induced by NO. Surprisingly, p53R2 is similarly upregulated by NO in two p53-deficient cell lines, showing the existence of p53-independent regulatory mechanisms. Delta Np73, which is overexpressed in many cancers, inhibits the transcriptional activity of p53 and p53 homologs. In p53-wt cells, the Delta Np73alpha isoform inhibits basal and NO-induced p53R2 protein expression. In p53-null cells, it also strongly inhibits p53R2 expression, and represses the enhancer activity of the p53-responsive element present in the p53R2-encoding gene. These results demonstrate that p53R2 expression can be controlled by p53 homologs in the absence of p53, and is downregulated by oncogenic Delta Np73 isoforms. Knocking down p53R2 in p53-wt cells dramatically enhances NO-induced DNA damages, indicating a protective function of the p53R2 ribonucleotide reductase subunit in prevention or repair of NO-mediated genotoxic injury.