Phosphorylation of casein kinase II by p34cdc2 in vitro and at mitosis.
In human epidermal carcinoma A431 cells, the beta subunit of casein kinase II is phosphorylated at an autophosphorylation site and at serine 209 which can be phosphorylated in vitro by p34cdc2 (Litchfield, D. W., Lozeman, F. J., Cicirelli, M. F., Harrylock, M., Ericsson, L. H., Piening, C. J., and Krebs, E. G. (1991) J. Biol. Chem. 266, 20380-20389). Given the importance of p34cdc2 in the regulation of cell cycle events, we were interested in examining the phosphorylation of casein kinase II during different stages of the cell cycle. In this study it is demonstrated that the extent of phosphorylation of serine 209 in the beta subunit is significantly increased relative to phosphorylation of the autophosphorylation site when chicken bursal lymphoma BK3A cells are arrested at mitosis by nocodazole treatment. This result suggests that serine 209 is a likely physiological target for p34cdc2. In addition, the alpha subunit of casein kinase II also undergoes dramatic phosphorylation with an associated alteration in its electrophoretic mobility when BK3A cells or human Jurkat cells are arrested with nocodazole. Phosphopeptide mapping studies indicate that p34cdc2 can phosphorylate in vitro the same peptides on the alpha subunit that are phosphorylated in cells arrested at mitosis. These phosphorylation sites were localized to serine and threonine residues in the carboxyl-terminal domain of alpha. Taken together, the results of this study indicate that casein kinase II is a probable physiological substrate for p34cdc2 and suggest that its functional properties could be affected in a cell cycle-dependent manner.
Litchfield
D W
DW
Department of Pharmacology, University of Washington, Seattle 98195.
Lüscher
B
B
Lozeman
F J
FJ
Eisenman
R N
RN
Krebs
E G
EG
D000818:Animals; D000906:Antibodies; D016203:CDC2 Protein Kinase / Q000378:metabolism*; D002294:Carcinoma, Squamous Cell; D047390:Casein Kinase II; D002417:Cattle; D002460:Cell Line; D002645:Chickens; D002852:Chromatography, Ion Exchange; D006801:Humans; D008223:Lymphoma; D046911:Macromolecular Substances; D008297:Male; D008938:Mitosis / Q000502:physiology*; D010449:Peptide Mapping; D010748:Phosphopeptides / Q000302:isolation & purification; D010766:Phosphorylation; D017346:Protein Serine-Threonine Kinases / Q000302:isolation & purification / Q000378:metabolism*; D013737:Testis / Q000201:enzymology; D014357:Trypsin
D000906:Antibodies; D046911:Macromolecular Substances; D010748:Phosphopeptides; D047390:Casein Kinase II; D017346:Protein Serine-Threonine Kinases; D016203:CDC2 Protein Kinase; D014357:Trypsin