The terminal region of beta-catenin promotes stability by shielding the Armadillo repeats from the axin-scaffold destruction complex

J Biol Chem. 2009 Oct 9;284(41):28222-28231. doi: 10.1074/jbc.M109.045039. Epub 2009 Aug 25.

Abstract

Post-translational stabilization of beta-catenin is a key step in Wnt signaling, but the features of beta-catenin required for stabilization are incompletely understood. We show that forms of beta-catenin lacking the unstructured C-terminal domain (CTD) show faster turnover than full-length or minimally truncated beta-catenins. Mutants that exhibit faster turnover show enhanced association with axin in co-transfected cells, and excess CTD polypeptide can compete binding of the beta-catenin armadillo (arm) repeat domain to axin in vitro, indicating that the CTD may restrict beta-catenin binding to the axin-scaffold complex. Fluorescent resonance energy transmission (FRET) analysis of cyan fluorescent protein (CFP)-arm-CTD-yellow fluorescent protein beta-catenin reveals that the CTD of beta-catenin can become spatially close to the N-terminal arm repeat region of beta-catenin. FRET activity is strongly diminished by the coexpression of beta-catenin binding partners, indicating that an unliganded groove is absolutely required for an orientation that allows FRET. Amino acids 733-759 are critical for beta-catenin FRET activity and stability. These data indicate that an N-terminal orientation of the CTD is required for beta-catenin stabilization and suggest a model where the CTD extends toward the N-terminal arm repeats, shielding these repeats from the beta-catenin destruction complex.

Publication types

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

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Axin Protein
  • Cell Line
  • Fluorescence Resonance Energy Transfer
  • Humans
  • Models, Molecular
  • Molecular Sequence Data
  • Protein Structure, Tertiary*
  • Repressor Proteins / genetics
  • Repressor Proteins / metabolism*
  • Signal Transduction / physiology
  • Two-Hybrid System Techniques
  • Wnt Proteins / genetics
  • Wnt Proteins / metabolism
  • Zebrafish Proteins / chemistry
  • Zebrafish Proteins / genetics
  • Zebrafish Proteins / metabolism
  • beta Catenin / chemistry*
  • beta Catenin / genetics
  • beta Catenin / metabolism*

Substances

  • Axin Protein
  • Repressor Proteins
  • Wnt Proteins
  • Zebrafish Proteins
  • beta Catenin