Development of a stabilized form of the regulatory CK2beta subunit that inhibits cell proliferation.
A number of cancers are characterized by elevated expression of CK2 (formerly casein kinase II), which has been implicated as a key component in cell proliferation and transformation. Two lines of evidence, (a) deregulated expression of CK2 and (b) CK2beta ubiquitination and degradation of these in a proteasome-dependent manner prompted further investigation of the regulation of CK2beta protein stability. We demonstrate that mutating six surface-exposed lysine residues to arginine (6KR) to interfere with ubiquitin attachment can stabilize CK2beta. Examination of 6KR expression in cells revealed increased stability over time and increased its steady-state expression level compared with CK2beta. In cells, 6KR was no longer sensitive to proteasome inhibition but maintained an elevated expression level. In our studies, 6KR functioned as a normal CK2 regulatory subunit, because it participated in CK2beta dimerization, associated with catalytic subunits, was autophosphorylated, and formed active, stable CK2 tetramers. The physiological role of CK2beta stabilization was investigated in cell proliferation assays, which showed a significant decrease in proliferation in cells expressing 6KR compared with CK2beta. Overall, our results indicate that a stabilized form of CK2beta can be used to inhibit cell proliferation.
Regulatory Biology and Functional Genomics Research Group, Siebens-Drake Medical Research Institute, Department of Biochemistry, Schulich School of Medicine and Dentistry, University of Western Ontario, London, Ontario N6A 5C1, Canada.
Ashley C
AC
French
Bernhard
B
Luscher
David W
DW
Litchfield
D000595:Amino Acid Sequence; D000818:Animals; D019556:COS Cells; D047390:Casein Kinase II / Q000502:physiology*; D049109:Cell Proliferation; D002471:Cell Transformation, Neoplastic; D002522:Chlorocebus aethiops; D005786:Gene Expression Regulation; D006367:HeLa Cells; D006801:Humans; D008239:Lysine / Q000737:chemistry; D008969:Molecular Sequence Data; D046988:Proteasome Endopeptidase Complex / Q000378:metabolism; D061988:Proteasome Inhibitors; D017386:Sequence Homology, Amino Acid; D025801:Ubiquitin / Q000737:chemistry
D061988:Proteasome Inhibitors; D025801:Ubiquitin; D047390:Casein Kinase II; D046988:Proteasome Endopeptidase Complex; D008239:Lysine