The zinc finger mutation C417R of I-kappa B kinase gamma impairs lipopolysaccharide- and TNF-mediated NF-kappa B activation through inhibiting phosphorylation of the I-kappa B kinase beta activation loop.
The activation of the I-kappaB kinase (IKK) complex by TNF or LPS stimulates phosphorylation and degradation of I-kappaBalpha, leading to the nuclear translocation of NF-kappaB. The IKK complex is mainly composed of two catalytic subunits, IKKalpha and IKKbeta, and a chaperon subunit IKKgamma. Although IKKgamma does not have catalytic activity, it is essential for IKK activation induced by multiple stimuli. Importantly, the key residue cysteine 417 at the zinc finger domain of IKKgamma has been found to be mutated to arginine (IKKgammaC417R) in a human genetic disorder called the anhydrotic ectodermal dysplasia with immunodeficiency. To understand the underlying mechanisms of immunodeficiency, we examined whether the IKKgammaC417R mutant modified IKK activation and NF-kappaB transcription stimulated by LPS or TNF in human monocytes. We found that overexpression of IKKgammaC417R severely impaired LPS- and TNF-induced I-kappaBalpha phosphorylation and degradation in a dominant-negative fashion. Also, LPS- and TNF-induced NF-kappaB transcription was inhibited by IKKgammaC417R. The reconstitution of IKKgamma, but not IKKgammaC417R, in IKKgamma-deficient cells restored NF-kappaB signaling, indicating the zinc finger structure of IKKgamma plays a key role in IKK activation. Moreover, C417R mutation in IKKgamma abolished both LPS- and TNF-induced phosphorylation of the activation loop of IKKbeta. Collectively, our results indicated that the zinc finger structure of IKKgamma plays a key role in LPS- and TNF-induced NF-kappaB activation. The anhydrotic ectodermal dysplasia with immunodeficiency patients' immunodeficiency may be associated with NF-kappaB defect in response to bacterial stimulation.
Yang
Fan
F
Department of Biologic and Materials Sciences, School of Dentistry, University of Michigan, Ann Arbor, MI 48109, USA.
Yamashita
Junro
J
Tang
Eric
E
Wang
Hom-Lay
HL
Guan
Kunliang
K
Wang
Cun-Yu
CY
D021581:Active Transport, Cell Nucleus / Q000235:genetics; D001120:Arginine / Q000235:genetics; D045744:Cell Line, Tumor; D002467:Cell Nucleus / Q000201:enzymology / Q000235:genetics / Q000378:metabolism; D003545:Cysteine / Q000235:genetics; D004789:Enzyme Activation / Q000235:genetics; D005822:Genetic Vectors; D006801:Humans; D051550:I-kappa B Kinase; D019169:Jurkat Cells; D008070:Lipopolysaccharides / Q000037:antagonists & inhibitors* / Q000494:pharmacology; D009154:Mutation; D016328:NF-kappa B / Q000037:antagonists & inhibitors* / Q000378:metabolism; D010766:Phosphorylation; D017346:Protein Serine-Threonine Kinases / Q000037:antagonists & inhibitors* / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism; D017434:Protein Structure, Tertiary / Q000235:genetics; D021122:Protein Subunits / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D012190:Retroviridae / Q000235:genetics; D014162:Transfection; D014409:Tumor Necrosis Factor-alpha / Q000037:antagonists & inhibitors* / Q000494:pharmacology; D015854:Up-Regulation / Q000235:genetics; D016335:Zinc Fingers / Q000235:genetics*
D008070:Lipopolysaccharides; D016328:NF-kappa B; D021122:Protein Subunits; D014409:Tumor Necrosis Factor-alpha; D001120:Arginine; D017346:Protein Serine-Threonine Kinases; C496559:CHUK protein, human; D051550:I-kappa B Kinase; C496560:IKBKB protein, human; C496565:IKBKE protein, human; D003545:Cysteine