The p25rum1 is an inhibitor of Cdc2 kinase expressed in fission yeast and plays an important role in cell-cycle control. As its amino-acid sequence suggests that p25rum1 has putative phosphorylation sites for mitogen-activated protein kinase (MAPK), we investigated the ability of MAPK to phosphorylate p25rum1. Direct in vitro kinase assay using GST-fusion proteins of wild-type as well as various mutants of p25rum1 demonstrated that MAPK phosphorylates the N-terminal portion of p25rum1 and residues Thr13 and Ser19 are major phosphorylation sites for MAPK
