Nuclear localization and formation of beta-catenin-lymphoid enhancer factor 1 complexes are not sufficient for activation of gene expression

Mol Cell Biol. 1999 Jun;19(6):4503-15. doi: 10.1128/MCB.19.6.4503.

Abstract

In response to activation of the Wnt signaling pathway, beta-catenin accumulates in the nucleus, where it cooperates with LEF/TCF (for lymphoid enhancer factor and T-cell factor) transcription factors to activate gene expression. The mechanisms by which beta-catenin undergoes this shift in location and participates in activation of gene transcription are unknown. We demonstrate here that beta-catenin can be imported into the nucleus independently of LEF/TCF binding, and it may also be exported from nuclei. We have introduced a small deletion within beta-catenin (Delta19) that disrupts binding to LEF-1, E-cadherin, and APC but not axin. This Delta19 beta-catenin mutant localizes to the nucleus because it may not be efficiently sequestered in the cytoplasm. The nuclear localization of Delta19 definitively demonstrates that the mechanisms by which beta-catenin localizes in the nucleus are completely independent of LEF/TCF factors. beta-Catenin and LEF-1 complexes can activate reporter gene expression in a transformed T-lymphocyte cell line (Jurkat) but not in normal T lymphocytes, even though both factors are nuclear. Thus, localization of both factors to the nucleus is not sufficient for activation of gene expression. Excess beta-catenin can squelch reporter gene activation by LEF-1-beta-catenin complexes but not activation by the transcription factor VP16. Taken together, these data suggest that a third component is necessary for gene activation and that this third component may vary with cell type.

Publication types

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

MeSH terms

  • Animals
  • COS Cells
  • Cadherins / metabolism
  • Cell Line
  • Cell Nucleus / metabolism*
  • Cycloheximide / pharmacology
  • Cytoplasm / metabolism
  • Cytoskeletal Proteins / metabolism*
  • Cytoskeletal Proteins / physiology*
  • DNA-Binding Proteins / metabolism*
  • DNA-Binding Proteins / physiology*
  • Dactinomycin / pharmacology
  • Gene Expression Regulation*
  • Humans
  • Ionomycin / pharmacology
  • Jurkat Cells
  • Lymphocytes / metabolism
  • Lymphoid Enhancer-Binding Factor 1
  • Microscopy, Fluorescence
  • Models, Genetic
  • Mutagenesis
  • Plasmids
  • Precipitin Tests
  • Protein Synthesis Inhibitors / pharmacology
  • Recombinant Fusion Proteins
  • Tetradecanoylphorbol Acetate / pharmacology
  • Trans-Activators*
  • Transcription Factors / metabolism*
  • Transcription Factors / physiology*
  • Transcriptional Activation
  • beta Catenin

Substances

  • CTNNB1 protein, human
  • Cadherins
  • Cytoskeletal Proteins
  • DNA-Binding Proteins
  • LEF1 protein, human
  • Lymphoid Enhancer-Binding Factor 1
  • Protein Synthesis Inhibitors
  • Recombinant Fusion Proteins
  • Trans-Activators
  • Transcription Factors
  • beta Catenin
  • Dactinomycin
  • Ionomycin
  • Cycloheximide
  • Tetradecanoylphorbol Acetate