Nucleolar targeting of the fbw7 ubiquitin ligase by a pseudosubstrate and glycogen synthase kinase 3

Mol Cell Biol. 2011 Mar;31(6):1214-24. doi: 10.1128/MCB.01347-10. Epub 2011 Jan 10.

Abstract

E3 ubiquitin ligases catalyze protein degradation by the ubiquitin-proteasome system, and their activity is tightly controlled. One level of regulation involves subcellular localization, and the Fbw7 tumor suppressor exemplifies this type of control. Fbw7 is the substrate-binding component of an SCF ubiquitin ligase that degrades critical oncoproteins. Alternative splicing produces three Fbw7 protein isoforms that occupy distinct compartments: Fbw7α is nucleoplasmic, Fbw7β is cytoplasmic, and Fbw7γ is nucleolar. We found that cancer-associated Fbw7 mutations that disrupt substrate binding prevent Fbw7γ nucleolar localization, implicating a substrate-like interaction in nucleolar targeting. We identified EBNA1-binding protein 2 (Ebp2) as the critical nucleolar factor that directly mediates Fbw7 nucleolar targeting. Ebp2 binds to Fbw7 like a substrate, and this is mediated by an Ebp2 degron that is phosphorylated by glycogen synthase kinase 3. However, despite these canonical substrate-like interactions, Fbw7 binding is largely uncoupled from Ebp2 turnover in vivo. Ebp2 thus acts like a pseudosubstrate that directly recruits Fbw7 to nucleoli.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Carrier Proteins / metabolism
  • Cell Line
  • Cell Line, Tumor
  • Cell Nucleolus / metabolism*
  • Cells, Cultured
  • Glycogen Synthase Kinase 3 / metabolism*
  • Humans
  • Mutation
  • Neoplasms / genetics
  • Phosphorylation
  • Protein Binding
  • Protein Isoforms / analysis
  • Protein Isoforms / genetics
  • Protein Isoforms / metabolism
  • Protein Transport
  • RNA-Binding Proteins
  • SKP Cullin F-Box Protein Ligases / analysis*
  • SKP Cullin F-Box Protein Ligases / genetics
  • SKP Cullin F-Box Protein Ligases / metabolism*

Substances

  • Carrier Proteins
  • EBNA1BP2 protein, human
  • Protein Isoforms
  • RNA-Binding Proteins
  • SKP Cullin F-Box Protein Ligases
  • Glycogen Synthase Kinase 3