Phosphorylation by the protein kinase CK2 promotes calpain-mediated degradation of IkappaBalpha.
Rapid IkappaBalpha turnover has been implicated in the high basal NF-kappaB activity in WEHI 231 B immature IgM(+) B cells. Here we show that treatment of WEHI 231 cells with apigenin, a selective inhibitor of the protein kinase CK2, decreased the rate of IkappaBalpha turnover and nuclear levels of NF-kappaB. Turnover of IkappaBalpha in these cells is mediated in part by the protease calpain. Since both CK2 and calpain target the proline-glutamic acid-serine-threonine (PEST) domain, we investigated the role of CK2 in the degradation of IkappaBalpha by calpain using an in vitro phosphorylation/degradation assay. CK2 phosphorylation enhanced mu-calpain-mediated degradation of wild-type IkappaBalpha, but not of mutant 3CIkappaBalpha, with S283A, T291A, and T299A mutations in phosphorylation sites within the PEST domain. Roles for CK2 and calpain in IkappaBalpha turnover were similarly shown in CH31 immature and CH12 mature IgM(+) B cells, but not in A20 and M12 IgG(+) B cells. These findings demonstrate for the first time that CK2 phosphorylation of serine/threonine residues in the PEST domain promotes calpain-mediated degradation of IkappaBalpha and thereby increases basal NF-kappaB levels in IgM(+) B cells.
Shen
J
J
Department of Laboratory Medicine, Boston University Medical School, Boston, MA 02118, USA.
Channavajhala
P
P
Seldin
D C
DC
Sonenshein
G E
GE
D047310:Apigenin; D002154:Calpain / Q000502:physiology*; D047390:Casein Kinase II; D002478:Cells, Cultured; D004268:DNA-Binding Proteins / Q000378:metabolism*; D005419:Flavonoids / Q000494:pharmacology; D006801:Humans; D020672:I-kappa B Proteins; D000072000:NF-KappaB Inhibitor alpha; D010766:Phosphorylation; D017346:Protein Serine-Threonine Kinases / Q000037:antagonists & inhibitors / Q000502:physiology*
D004268:DNA-Binding Proteins; D005419:Flavonoids; D020672:I-kappa B Proteins; C000606204:NFKBIA protein, human; D000072000:NF-KappaB Inhibitor alpha; D047310:Apigenin; D047390:Casein Kinase II; D017346:Protein Serine-Threonine Kinases; D002154:Calpain