Inducible phosphorylation of NF-kappa B p65 at serine 468 by T cell costimulation is mediated by IKK epsilon.
Here we identify IKKepsilon as a novel NF-kappaB p65 kinase that mediates inducible phosphorylation of Ser468 and Ser536 in response to T cell costimulation. In addition, the kinase activity of IKKepsilon contributes to the control of p65 nuclear uptake. Serines 468 and 536 are evolutionarily conserved, and the surrounding amino acids display sequence homology. Down-regulation of IKKepsilon levels by small interfering RNA does not affect inducible phosphorylation of Ser536 but largely prevents Ser468 phosphorylation induced by T cell costimulation. Ser536-phosphorylated p65 is found predominantly in the cytosol. In contrast, the Ser468 phosphorylated form of this transcription factor occurs mainly in the nucleus, suggesting a function for transactivation. Reconstitution of p65-/- cells with either wild type p65 or point-mutated p65 variants showed that inducible phosphorylation of Ser468 serves to enhance p65-dependent transactivation. These results also provide a mechanistic link that helps to explain the relevance of IKKepsilon for the expression of a subset of NF-kappaB target genes without affecting cytosolic IkappaBalpha degradation.
Mattioli
Ivan
I
Department of Chemistry and Biochemistry, University of Bern, Freiestrasse 3, 3012 Bern, Switzerland.
Geng
Hui
H
Sebald
Andrea
A
Hodel
Michael
M
Bucher
Cyril
C
Kracht
Michael
M
Schmitz
M Lienhard
ML
D000595:Amino Acid Sequence; D000818:Animals; D002467:Cell Nucleus / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D051550:I-kappa B Kinase / Q000502:physiology*; D020672:I-kappa B Proteins / Q000378:metabolism; D019169:Jurkat Cells; D008213:Lymphocyte Activation / Q000502:physiology*; D051379:Mice; D008969:Molecular Sequence Data; D000072000:NF-KappaB Inhibitor alpha; D010766:Phosphorylation; D012694:Serine / Q000378:metabolism*; D013601:T-Lymphocytes / Q000378:metabolism*; D051996:Transcription Factor RelA / Q000378:metabolism*
D020672:I-kappa B Proteins; C000606204:NFKBIA protein, human; C000606205:Nfkbia protein, mouse; D051996:Transcription Factor RelA; D000072000:NF-KappaB Inhibitor alpha; D012694:Serine; D051550:I-kappa B Kinase