Necdin interacts with the Msx2 homeodomain protein via MAGE-D1 to promote myogenic differentiation of C2C12 cells

J Biol Chem. 2004 Sep 24;279(39):40484-93. doi: 10.1074/jbc.M404143200. Epub 2004 Jul 21.


Necdin is a potent growth suppressor that is expressed predominantly in postmitotic cells such as neurons and skeletal muscle cells. Necdin shows a significant homology to MAGE (melanoma antigen) family proteins, all of which contain a large homology domain. MAGE-D1 (NRAGE, Dlxin-1) interacts with the Dlx/Msx family homeodomain proteins via an interspersed hexapeptide repeat domain distinct from the homology domain. Here we report that necdin associates with the Msx homeodomain proteins via MAGE-D1 to modulate their function. In vitro binding and co-immunoprecipitation analyses revealed that MAGE-D1 directly interacted with necdin via the homology domain and Msx1 (or Msx2) via the repeat domain. A ternary complex of necdin, MAGE-D1, and Msx2 was formed in vitro, and an endogenous complex containing these three proteins was detected in differentiating embryonal carcinoma cells. Co-expression of necdin and MAGE-D1 released Msx-dependent transcriptional repression. C2C12 myoblast cells that were stably transfected with Msx2 cDNA showed a marked reduction in myogenic differentiation, and co-expression of necdin and MAGE-D1 canceled the Msx2-dependent repression. These results suggest that necdin and MAGE-D1 cooperate to modulate the function of Dlx/Msx homeodomain proteins in cellular differentiation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Blotting, Western
  • COS Cells
  • Cell Differentiation
  • Cell Line
  • Cell Line, Tumor
  • DNA, Complementary / metabolism
  • DNA-Binding Proteins / metabolism*
  • Gene Deletion
  • Genetic Vectors
  • Homeodomain Proteins
  • Immunoblotting
  • Immunohistochemistry
  • Mice
  • Mice, Inbred ICR
  • Microscopy, Fluorescence
  • Mitosis
  • Models, Biological
  • Neoplasm Proteins / metabolism
  • Nerve Tissue Proteins / physiology*
  • Neurons / metabolism
  • Nuclear Proteins / physiology*
  • Precipitin Tests
  • Protein Binding
  • Protein Structure, Tertiary
  • Tissue Distribution
  • Transcription, Genetic


  • DNA, Complementary
  • DNA-Binding Proteins
  • Homeodomain Proteins
  • MSX2 protein
  • Maged1 protein, mouse
  • Neoplasm Proteins
  • Nerve Tissue Proteins
  • Nuclear Proteins
  • necdin