Staurosporine induces tyrosine phosphorylation of a 145 kDa protein but does not activate gp140^trk in PC12 cells

Straurosporine, a protein kinase C inhibitor, induces neurite outgrowth in PC12 cells similarly to nerve growth factor (NGF). Since NGF neurotropic effects are transduced by the 'trk' gene product 140 kDa tyrosine kinase receptor, gp140^trk, we investigated the role of gp140^trk and tyrosine phosphorylations in staurospine neurotropic effects. A direct correlation between staurosporine neurotropic effects and a novel stimulation of tyrosine phosphorylation of a 145 kDa protein (p145) with the following characteristics has been discovered: (1) Staurosporine specifically induced, among indolcarbazoles-K252a derivatives, in a dose-dependent manner (5-100 nM), p145 tyrosine phosphorylation and neurite outgrowth. (2) Staurosporine-induced p145 tyrosine phosphorylation was selective compared to other neurotropic compounds such as 8-Br-cAMP, acidic and basic fibroblast growth factors and NGF. (3) Staurosporine stimulation of p145 tyrosine phosphorylation gradually increased during the first 8 h of staurosporine treatment coinciding with the initiation of neurotropic effect. (4) K252a, a selective inhibitor of NGF actions, and several tyrphostins did not block staurosporine-induced p 145 tyrosine phosphorylation and neurotropic effects. (5) Staurosporine stimulation of p145 tyrosine phosphorylation and neurotropic effects are independent of PKC. (6) Staurosporine did not activate gp140^trk-NGF receptor in PC12 cells. The present study proposes staurosporine as a pharmacological tool to study the role of tyrosine phosphorylation pathway(s), such as p145 phosphorylation, in the action neurotropic agents.