Serine 32 and serine 36 of IkappaBalpha are directly phosphorylated by protein kinase CKII in vitro.
IkappaBalpha is an inherently unstable protein which binds to and retains the ubiquitous transcription factor NFkappaB in the cytoplasm of resting cells. A continuous low level translocation of NFkappaB to the nucleus, secondary to the basal turnover of IkappaBalpha, is hypothesized to be necessary for cellular maturation, survival and, potentially, transformation. In response to cellular stimulation by inflammatory cytokines or mitogens, IkappaBalpha is rapidly degraded allowing larger pools of NFkappaB to translocate to the nucleus. Phosphorylation of IkappaBalpha at serine 32 (S32) and serine 36 (S36) is necessary for this stimuli-induced degradation. IKKalpha/beta kinases and p90(rsk1)are involved in stimuli-induced targeting of one or both of these IkappaBalpha sites. Whether other kinases phosphorylate S32 and S36 directly, and if so, what function they serve in NFkappaB activation remains unknown. Here we present evidence of a direct phosphorylation of IkappaBalpha at both S32 and S36 by purified or immunoprecipitated protein kinase CKII (PK-CKII) and a specific in vivo association between IkappaBalpha and PK-CKII. This PK-CKII-specific kinase activity is not found within the IKKalpha/beta-containing signalsome complex and is biochemically distinct from that of the IKKalpha/beta kinases. The identification of an additional N-terminal IkappaBalpha kinase which is constitutively active and not significantly inducible raises numerous possibilities as to its role in cellular function.
Department of Immunology, Division of Infectious Diseases, Mayo Clinic,Rochester, MN 55905, USA.
J A
JA
Taylor
G D
GD
Bren
K N
KN
Pennington
S A
SA
Trushin
S
S
Asin
C V
CV
Paya
D019794:2,3-Diphosphoglycerate / Q000494:pharmacology; D000818:Animals; D047390:Casein Kinase II; D004268:DNA-Binding Proteins / Q000378:metabolism*; D006493:Heparin / Q000494:pharmacology; D006801:Humans; D051550:I-kappa B Kinase; D020672:I-kappa B Proteins; D000072000:NF-KappaB Inhibitor alpha; D010766:Phosphorylation; D011233:Precipitin Tests; D011493:Protein Kinase C / Q000494:pharmacology; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism*; D051381:Rats; D012694:Serine / Q000502:physiology*; D020298:U937 Cells
D004268:DNA-Binding Proteins; D020672:I-kappa B Proteins; C000606204:NFKBIA protein, human; C000606206:Nfkbia protein, rat; D019794:2,3-Diphosphoglycerate; D000072000:NF-KappaB Inhibitor alpha; D012694:Serine; D006493:Heparin; D047390:Casein Kinase II; D017346:Protein Serine-Threonine Kinases; C496559:CHUK protein, human; D051550:I-kappa B Kinase; C496560:IKBKB protein, human; C496565:IKBKE protein, human; D011493:Protein Kinase C