Genetic evidence for the essential role of beta-transducin repeat-containing protein in the inducible processing of NF-kappa B2/p100

J Biol Chem. 2002 Jun 21;277(25):22111-4. doi: 10.1074/jbc.C200151200. Epub 2002 May 6.

Abstract

Processing of the nf kappa b2 gene product p100 to generate p52 is an important step in NF-kappa B regulation. This step is regulated by a nonclassical NF-kappa B signaling pathway involving the NF-kappa B-inducing kinase (NIK). NIK induces p100 processing by triggering phosphorylation of specific C-terminal serines of p100. However, the downstream molecular events leading to p100 processing remain unclear. Here we show that NIK induced the physical recruitment of beta-transducin repeat-containing protein (beta-TrCP), a component of the SCF ubiquitin ligase complex, to p100. This event required the phosphorylation sites as well as the death domain of p100. Using the RNA interference technique, we demonstrated that beta-TrCP is essential for NIK-induced p100 ubiquitination and processing. Interestingly the constitutive processing of p100 mutants was independent of beta-TrCP. These results suggest that beta-TrCP is an essential component of NIK-induced p100 processing.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Amino Acid Sequence
  • Cell Line
  • DNA, Complementary / metabolism
  • Humans
  • Immunoblotting
  • Jurkat Cells
  • Molecular Sequence Data
  • Mutation
  • NF-kappa B / chemistry*
  • NF-kappa B / metabolism
  • NF-kappa B p52 Subunit
  • Peptide Synthases / chemistry*
  • Peptide Synthases / genetics
  • Peptide Synthases / metabolism*
  • Precipitin Tests
  • Protein Binding
  • Protein Structure, Tertiary
  • Reverse Transcriptase Polymerase Chain Reaction
  • SKP Cullin F-Box Protein Ligases
  • Sequence Homology, Amino Acid
  • Signal Transduction
  • Stem Cell Factor / metabolism
  • Transducin / chemistry*
  • Transfection

Substances

  • DNA, Complementary
  • NF-kappa B
  • NF-kappa B p52 Subunit
  • Stem Cell Factor
  • SKP Cullin F-Box Protein Ligases
  • Transducin
  • Peptide Synthases