Raf-1-associated protein phosphatase 2A as a positive regulator of kinase activation

The Raf-1 kinase plays a key role in relaying proliferation signals elicited by mitogens or oncogenes, Raf-1 is regulated by complex and incompletely understood mechanisms including phosphorylation, A number of studies have indicated that phosphorylation of serines 259 and 621 can inhibit the Raf-1 kinase, We show that both serines are hypophosphorylated during early mitogenic stimulation and that hypophosphorylation correlates with peak Raf-1 activation. Concentrations of okadaic acid that selectively inhibit protein phosphatase 2A (PP2A) induce phosphorylation of these residues and prevent maximal activation of the Raf-1 kinase. This effect is mediated via phosphorylation of serine 259. The PP2A core heterodimer forms complexes with Raf-1 in vivo and in vitro. These data identify PP2A as a positive regulator of Raf-1 activation and are the first indication that PP2A may support the activation of an associated kinase.