Crosstalk between TAp73 and TGF-β in fibroblast regulates iNOS expression and Nrf2-dependent gene transcription

Inducible nitric oxide synthase (iNOS) activity produces anti-tumor and anti-microbial effects but also promotes carcinogenesis through mutagenic, immunosuppressive and pro-angiogenic mechanisms. The tumor suppressor p53 contributes to iNOS downregulation by repressing induction of the NOS2 gene encoding iNOS, thereby limiting NO-mediated DNA damages. This study focuses on the role of the p53 homologue TAp73 in the regulation of iNOS expression. Induction of iNOS by immunological stimuli was upregulated in immortalized MEFs from TAp73−/− mice, compared to TAp73+/+ fibroblasts. This overexpression resulted both from increased levels of NOS2 transcripts, and from an increased stability of the protein. Limitation of iNOS expression by TAp73 in wild-type cells is alleviated by TGF-β receptor I inhibitors, suggesting a cooperation between TAp73 and TGF-β in suppression of iNOS expression. Accordingly, downregulation of iNOS expression by exogenous TGF-β1 was impaired in TAp73−/− fibroblasts. Increased NO production in these cells resulted in a stronger, NO-dependent induction of Nrf2 target genes, indicating that the Nrf2-dependent adaptive response to nitrosative stress in fibroblasts is proportional to iNOS activity. NO-dependent induction of two HIF-1 target genes was also stronger in TAp73-deficient cells. Finally, the antimicrobial action of NO against Trypanosoma musculi parasites was enhanced in TAp73−/− fibroblasts. Our data indicate that tumor suppressive TAp73 isoforms cooperate with TGF-β to control iNOS expression, NO-dependent adaptive responses to stress, and pathogen proliferation. [Display omitted] •Cytokine-induced iNOS expression is increased in TAp73-deficient fibroblasts.•TAp73 cooperates with TGF-β to suppress iNOS transcription and protein expression.•Nrf2-dependent induction of antioxidant genes is enhanced in TAp73 −/− fibroblasts.•NO-dependent upregulation of HIF-1 target genes is stronger in TAp73 −/− fibroblasts.•TAp73 −/− fibroblasts exhibit a higher NO-mediated antimicrobial activity.