Characterization and Physiological Importance of a Novel Cell Cycle Regulated Protein Kinase in Xenopus laevisOocytes That Phosphorylates Cyclin B2

Abstract

We have partially purified a specific cyclin B2 kinase (cyk) from prophase oocytes of Xenopus laevisafter an ATP-γ-S activation step. Phosphopeptide analysis identified Ser53 as the major in vitrophosphorylation site for cyk in cyclin B2. Using a synthetic peptide derived from cyclin B2 encompassing Ser53 (cyktide) as a substrate, cyk was shown to be activated during progesterone-induced maturation, with a peak of activity between 40 and 50% maturation. A sustained high cyk activity was observed in oscillating egg extracts. Microinjection of cyk-phosphorylated cyclin B2 into prophase oocytes accelerated progesterone-induced maturation by about 2 h, indicating that cyclin B2 is a relevant substrate for cyk and that the function of cyk is situated upstream of cdc2-cyclin B activation. Microinjection of cyk-phosphorylated cyktide or a combination of cyk and cyclin B1 into G 2fibroblasts induced significant changes in cell morphology, reminiscent of a premature prophase-like phenotype. Similarly, addition of cyk-phosphorylated cyktide in cyclin B1-dependent interphase extracts resulted in histone H1 kinase activation.