Different kinases phosphorylate nucleophosmin/B23 at different sites during G₂and M phases of the cell cycle

The recombinant GST-nucleophosmin/B23 and the truncated mutants were tested for phosphorylation in cell-free extracts of G2and M phases or by purified kinases. Our results indicated that a threonine residue at amino acids (a.a.) 185-240 was phosphorylated by cdc2 kinase during the entry of mitosis while the serine phosphorylation site at the middle acidic portion of the molecule (a.a. 83-152) was phosphorylated by casein kinase II during G2phase. Our results also showed that there was possibly another serine phosphorylation at site other than the middle portion of nucleophosmin/B23 (a.a. 83-152) during the entry of cells into mitosis. The demonstration of the characteristic changes in phosphorylation of nucleophosmin/B23 during the cell cycle implicates important role of nucleophosmin/B23 in the control of the fate of nucleoli and cell growth.