beta-TrCP binding and processing of NF-kappaB2/p100 involve its phosphorylation at serines 866 and 870.
Processing of the NF-kappaB2 precursor protein p100 is a major step in noncanonical NF-kappaB signaling. This signaling step requires the NF-kappaB inducing kinase (NIK) and its downstream kinase, IkappaB kinase alpha (IKKalpha). We show here that p100 undergoes phosphorylation at serines 866, 870, and possibly 872, in cells stimulated with noncanonical NF-kappaB stimuli or transfected with NIK and IKKalpha. Phosphorylation of this serine cluster creates a binding site for beta-TrCP, the receptor subunit of the beta-TrCP(SCF) ubiquitin ligase. Mutation of either serine 866 or serine 870 abolishes the beta-TrCP recruitment and ubiquitination of p100. The functional significance of p100 phosphorylation is further supported by the finding that this molecular event occurs in a NIK- and IKKalpha-dependent manner. Additionally, induction of p100 phosphorylation can be blocked by a protein synthesis inhibitor, suggesting the requirement of de novo protein synthesis. These data suggest that p100 processing involves its phosphorylation at specific terminal serines, which form a binding site for beta-TrCP thereby regulating p100 ubiquitination.
Liang
Chunyang
C
Department of Microbiology and Immunology, Pennsylvania State University College of Medicine, Hershey Medical Center, P.O. Box 850, Hershey, Pennsylvania 17033, USA.
Zhang
Minying
M
Sun
Shao-Cong
SC
D000818:Animals; D001665:Binding Sites / Q000235:genetics; D006801:Humans; D051550:I-kappa B Kinase / Q000378:metabolism; D051379:Mice; D009154:Mutation / Q000235:genetics; D052003:NF-kappa B p52 Subunit / Q000378:metabolism*; D010768:Phosphoserine / Q000378:metabolism*; D011485:Protein Binding; D014176:Protein Biosynthesis; D011499:Protein Processing, Post-Translational; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism; D044844:beta-Transducin Repeat-Containing Proteins / Q000378:metabolism*; D000097782:NF-kappaB-Inducing Kinase
D051550:I-kappa B Kinase; D052003:NF-kappa B p52 Subunit; D010768:Phosphoserine; D017346:Protein Serine-Threonine Kinases; D044844:beta-Transducin Repeat-Containing Proteins; D000097782:NF-kappaB-Inducing Kinase