Evidence for regulation of mitotic progression through temporal phosphorylation and dephosphorylation of CK2alpha.
Proper mitotic progression is crucial for maintenance of genomic integrity in proliferating cells and is regulated through an intricate series of events, including protein phosphorylation governed by a complex network of protein kinases. One kinase family implicated in the regulation of mitotic progression is protein kinase CK2, a small family of enzymes that is overexpressed in cancer and induces transformation in mice and cultured fibroblasts. CK2alpha, one isoform of the catalytic subunits of CK2, is maximally phosphorylated at four sites in nocodazole-treated cells. To investigate the effects of CK2alpha phosphorylation on mitotic progression, we generated phosphospecific antibodies against its mitotic phosphorylation sites. In U2OS cells released from S-phase arrest, these antibodies reveal that CK2alpha is most highly phosphorylated in prophase and metaphase. Phosphorylation gradually decreases during anaphase and becomes undetectable during telophase and cytokinesis. Stable expression of phosphomimetic CK2alpha (CK2alpha-4D, CK2alpha-4E) results in aberrant centrosome amplification and chromosomal segregation defects and loss of mitotic cells through mitotic catastrophe. Conversely, cells expressing nonphosphorylatable CK2alpha (CK2alpha-4A) show a decreased ability to arrest in mitosis following nocodazole treatment, suggesting involvement in the spindle assembly checkpoint. Collectively, these studies indicate that reversible phosphorylation of CK2alpha requires precise regulation to allow proper mitotic progression.
St-Denis
Nicole A
NA
Department of Biochemistry, Schulich School of Medicine and Dentistry, University of Western Ontario, London, Ontario, Canada.
Derksen
D Richard
DR
Litchfield
David W
DW
D000818:Animals; D001665:Binding Sites / Q000276:immunology; D047390:Casein Kinase II / Q000378:metabolism* / Q000502:physiology; D002460:Cell Line; D048750:Cell Nucleus Division; D018385:Centrosome; D020090:Chromosome Segregation; D048749:Cytokinesis; D051379:Mice; D008938:Mitosis; D010766:Phosphorylation / Q000502:physiology; D008941:Spindle Apparatus
D047390:Casein Kinase II