Several serine/threonine protein kinases have been implicated as intermediates in signal transduction pathways. These include ERK/MAP kinases, ribosomal S6 kinase (Rsk) and Raf-1. Raf-1 is a cytoplasmic protein with intrinsic serine/threonine activity. It is broadly expressed in nearly all cell lines tested to date and is the cellular homolog of v-Raf, the product of the transforming gene of the 3611 strain of murine sarcoma virus. The unregulated kinase activity of the v-Raf protein has been associated with transformation and mitogenesis while the activity of Raf-1 is normally suppressed by a regulatory N-terminal domain. A-Raf, a second member of the Raf gene family of serine/ threonine protein kinases, exhibits substantial homology to Raf-1 within the kinase domain of the two molecules, but less homology elsewhere. Expression of A-Raf is found at highest levels in urogenital tissues and kidney and at lowest level in brain tissue.