Induction of p100 processing by NF-kappaB-inducing kinase involves docking IkappaB kinase alpha (IKKalpha) to p100 and IKKalpha-mediated phosphorylation.
The processing of the nfkappab2 gene product p100 to generate p52 is a regulated event, which is important for the instrumental function of NF-kappaB. We previously demonstrated that this tightly controlled event is regulated positively by NF-kappaB-inducing kinase (NIK) and its downstream kinase, IkappaB kinase alpha (IKKalpha). However, the precise mechanisms by which NIK and IKKalpha induce p100 processing remain unclear. Here, we show that, besides activating IKKalpha, NIK also serves as a docking molecule recruiting IKKalpha to p100. This novel function of NIK requires two specific amino acid residues, serine 866 and serine 870, of p100 that are known to be essential for inducible processing of p100. We also show that, after being recruited into p100 complex, activated IKKalpha phosphorylates specific serines located in both N- and C-terminal regions of p100 (serines 99, 108, 115, 123, and 872). The phosphorylation of these specific serines is the prerequisite for ubiquitination and subsequent processing of p100 mediated by the beta-TrCP ubiquitin ligase and 26 S proteasome, respectively. These results highlight the critical but different roles of NIK and IKKalpha in regulating p100 processing and shed light on the mechanisms mediating the tight control of p100 processing. These data also provide the first evidence for explaining why overexpression of IKKalpha or its activation by many other stimuli such as tumor necrosis factor and mitogens fails to induce p100 processing.
Xiao
Gutian
G
Department of Microbiology and Immunology, Pennsylvania State University College of Medicine, Hershey, Pennsylvania 17033, USA. xiao@biology.rutgers.edu
Fong
Abraham
A
Sun
Shao-Cong
SC
D001665:Binding Sites; D045744:Cell Line, Tumor; D005822:Genetic Vectors; D006801:Humans; D051550:I-kappa B Kinase; D016328:NF-kappa B / Q000737:chemistry / Q000378:metabolism*; D052003:NF-kappa B p52 Subunit; D010447:Peptide Hydrolases / Q000378:metabolism; D010766:Phosphorylation; D011233:Precipitin Tests; D046988:Proteasome Endopeptidase Complex; D011485:Protein Binding; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D012694:Serine / Q000737:chemistry; D014162:Transfection; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism; D025801:Ubiquitin / Q000378:metabolism; D044767:Ubiquitin-Protein Ligases / Q000378:metabolism; D044844:beta-Transducin Repeat-Containing Proteins / Q000737:chemistry* / Q000378:metabolism; D000097782:NF-kappaB-Inducing Kinase
D051550:I-kappa B Kinase; D016328:NF-kappa B; D052003:NF-kappa B p52 Subunit; D010447:Peptide Hydrolases; D046988:Proteasome Endopeptidase Complex; D017346:Protein Serine-Threonine Kinases; D012694:Serine; D014409:Tumor Necrosis Factor-alpha; D025801:Ubiquitin; D044767:Ubiquitin-Protein Ligases; D044844:beta-Transducin Repeat-Containing Proteins; D000097782:NF-kappaB-Inducing Kinase; C496559:CHUK protein, human; C496560:IKBKB protein, human; C496565:IKBKE protein, human; C061553:ATP dependent 26S protease