Peroxynitrite induces an alternative NF-kappaB activation pathway in L8 rat myoblasts

The role of peroxynitrite in NF-kappaB activation remains controversial. This study investigated NF-kappaB activation by peroxynitrite in skeletal myocytes. Myocytes were treated with NO and peroxynitrite donors. Both NO and peroxynitrite caused NF-kappaB activation (measured by p65 nuclear translocation and luciferase expression). NO donor-induced NF-kappaB activation was transient, dependent on I-kappaB alpha degradation, and was decreased in the presence of I-kappaB alpha super-repressor. Conversely, peroxynitrite donors induced NF-kappaB activation that was dependent on tyrosine nitration of I-kappaB alpha, but independent of its serine phosphorylation and degradation. This activation did not decrease in the presence of I-kappaB alpha super-repressor. Prolonged exposure to peroxynitrite resulted in nontransient NF-kappaB activation and high iNOS expression. Proteasome inhibitor MG-132 did not diminish SIN-1-induced NF-kappaB activation. Tyrosine nitration inhibitor EGCG re-established transient NF-kappaB activation with I-kappaB alpha degradation after SIN-1 treatment. EGCG, but not MG-132 decreased SIN-1- dependent iNOS expression. Peroxynitrite activates NF-kappaB in skeletal myocytes through an alternative mechanism, in which I-kappaB alpha is nitrated on tyrosine and dissociated from NF-kappaB, thus enabling its nontransient activation. This resulted in prolonged iNOS expression. Hence, peroxynitrite may exacerbate inflammatory responses mediated by NF-kappaB.