Activation of Mil/Raf protein kinases in mitotic cells

The c-Raf-1 protein kinase is a major element of several signal transduction pathways and thought to be involved in entry into the S phase of the cell cycle. Here we show that c-Raf-1 as well as the transforming viral fusion protein Gag-Mil, in which most of the amino terminal regulatory region of the avian Raf homologue Mil is deleted, are activated five- to sixfold in mitotic cells. Mitotic activation of Mil/Raf kinase activity correlates with reduced electrophoretic mobility caused by hyperphosphorylation at serine/threonine residues located in the carboxy terminal part of c-Raf-1. Mitotic hyperphosphorylation occurs in various cell-lines indicating that it is ubiquitous. Our data suggest a novel function for Mil/Raf kinases in late stages of the cell cycle.