Persistent activation of RelA by respiratory syncytial virus involves protein kinase C, underphosphorylated IκBβ, and sequestration of protein phosphatase 2A by the viral phosphoprotein

Respiratory syncytial virus (RSV) activated the RelA (p65) subunit of nuclear factor kappa B (NF-kappa B) over many hours postinfection. The initial activation coincided with phosphorylation and degradation of I kappa B alpha, the cytoplasmic inhibitor of RelA, During persistent activation of NF-kappa B at later times in infection, syntheses of inhibitors I kappa B beta as well as I kappa B beta were restored. However, the resynthesized I kappa B beta was in an underphosphorylated state, which apparently prevented inhibition of NF-kappa B. Use of specific inhibitors suggested that the pathway leading to the persistent-but not the initial-activation of NF-kappa B involved signaling through protein kinase C (PKC) and reactive oxygen intermediates of nonmitochondrial origin, whereas phospholipase C or D played little or no role. Thus, RSV infection led to the activation of NF-kappa B by a biphasic mechanism: a transient or early activation involving phosphorylation of the inhibitor I kappa B polypeptides, and a persistent or long-term activation requiring PKC and the generation of hypophosphorylated I kappa B beta. At least a part of the activation was through a novel mechanism in which the viral phosphoprotein P associated with but was not dephosphorylated by protein phosphatase 2A and thus sequestered and inhibited the latter. We postulate that this led to a net increase in the phosphorylation state of signaling proteins that are responsible for RelA activation.