Erythropoietin-Stimulated Raf-1 Tyrosine Phosphorylation Is Associated with the Tyrosine Kinase Lyn in J2E Erythroleukemic Cells

The serine/threonine kinase Raf-1 is crucial for transducing intracellular signals emanating from numerous growth factors. Here we used the J2E erythroid cell line transformed by the ν-raf/ν-myc oncogenes to examine the effects of erythropoietin on endogenous Raf-1 activity. Despite the presence of constitutively active v-raf in these cells, Raf-1 exokinase activity increased after erythropoietin stimulation. This increase in enzymatic activity coincided with tyrosine phosphorylation of Raf-1 on residue Y341. Significantly, the tyrosine kinase Lyn coimmunoprecipitated with Raf-1, and Raf-1 was not tyrosine-phosphorylated in a J2E subclone lacking Lyn. Therefore, it was concluded that Lyn may be the kinase responsible for tyrosine phosphorylating Raf-1 and increasing its exokinase activity in response to erythropoietin.