Constitutive nuclear expression of the I kappa B kinase complex and its activation in human neutrophils.
A singular feature of human neutrophils is that they constitutively express substantial amounts of NF-kappaB/Rel proteins and IkappaB-alpha in the nucleus. In this study, we show that in these cells, IkappaB kinase alpha (IKKalpha), IKKbeta, and IKKgamma also partially localize to the nucleus, whereas IKK-related kinases (IKKepsilon, TANK-binding kinase-1) are strictly cytoplasmic, and the NF-kappaB-inducing kinase is strictly nuclear. Following neutrophil activation, IKKbeta and IKKgamma become transiently phosphorylated in both the cytoplasm and nucleus, whereas IKKalpha transiently vanishes from both compartments in what appears to be an IKKbeta-dependent process. These responses are paralleled by the degradation of IkappaB-alpha, and by the phosphorylation of RelA on serine 536, in both compartments. Although both proteins can be IKK substrates, inhibition of IKK prevented IkappaB-alpha phosphorylation, while that of RelA was mostly unaffected. Finally, we provide evidence that the nuclear IKK isoforms (alpha, beta, gamma) associate with chromatin following neutrophil activation, which suggests a potential role in gene regulation. This is the first study to document IKK activation and the phosphorylation of NF-kappaB/Rel proteins in primary neutrophils. More importantly, our findings unveil a hitherto unsuspected mode of activation for the IKK/IkappaB signaling cascade within the cell nucleus.
Ear
Thornin
T
Pulmonary Division, Faculty of Medicine, Université de Sherbrooke, Sherbrooke, Québec, Canada.
Cloutier
Alexandre
A
McDonald
Patrick P
PP
D021581:Active Transport, Cell Nucleus / Q000502:physiology; D002478:Cells, Cultured; D002843:Chromatin / Q000201:enzymology; D003593:Cytoplasm / Q000201:enzymology; D004789:Enzyme Activation / Q000235:genetics; D006801:Humans; D051550:I-kappa B Kinase; D007527:Isoenzymes / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism; D016328:NF-kappa B / Q000378:metabolism; D018375:Neutrophil Activation / Q000235:genetics; D009504:Neutrophils / Q000166:cytology / Q000201:enzymology* / Q000378:metabolism; D009687:Nuclear Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism; D010766:Phosphorylation; D017346:Protein Serine-Threonine Kinases / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism; D016192:Resting Phase, Cell Cycle / Q000502:physiology; D015398:Signal Transduction / Q000235:genetics / Q000502:physiology; D013379:Substrate Specificity / Q000502:physiology; D051996:Transcription Factor RelA
D002843:Chromatin; D007527:Isoenzymes; D016328:NF-kappa B; D009687:Nuclear Proteins; D051996:Transcription Factor RelA; D017346:Protein Serine-Threonine Kinases; C496559:CHUK protein, human; D051550:I-kappa B Kinase; C496560:IKBKB protein, human; C496565:IKBKE protein, human