A direct physical interaction between Nanog and Sox2 regulates embryonic stem cell self-renewal

EMBO J. 2013 Aug 14;32(16):2231-47. doi: 10.1038/emboj.2013.161. Epub 2013 Jul 26.


Embryonic stem (ES) cell self-renewal efficiency is determined by the Nanog protein level. However, the protein partners of Nanog that function to direct self-renewal are unclear. Here, we identify a Nanog interactome of over 130 proteins including transcription factors, chromatin modifying complexes, phosphorylation and ubiquitination enzymes, basal transcriptional machinery members, and RNA processing factors. Sox2 was identified as a robust interacting partner of Nanog. The purified Nanog-Sox2 complex identified a DNA recognition sequence present in multiple overlapping Nanog/Sox2 ChIP-Seq data sets. The Nanog tryptophan repeat region is necessary and sufficient for interaction with Sox2, with tryptophan residues required. In Sox2, tyrosine to alanine mutations within a triple-repeat motif (S X T/S Y) abrogates the Nanog-Sox2 interaction, alters expression of genes associated with the Nanog-Sox2 cognate sequence, and reduces the ability of Sox2 to rescue ES cell differentiation induced by endogenous Sox2 deletion. Substitution of the tyrosines with phenylalanine rescues both the Sox2-Nanog interaction and efficient self-renewal. These results suggest that aromatic stacking of Nanog tryptophans and Sox2 tyrosines mediates an interaction central to ES cell self-renewal.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Proliferation*
  • Colony-Forming Units Assay
  • Embryonic Stem Cells / metabolism
  • Embryonic Stem Cells / physiology*
  • Homeodomain Proteins / metabolism*
  • Immunoblotting
  • Immunoprecipitation
  • Mice
  • Nanog Homeobox Protein
  • Plasmids / genetics
  • Protein Interaction Domains and Motifs / genetics*
  • Protein Interaction Mapping
  • SELEX Aptamer Technique
  • SOXB1 Transcription Factors / metabolism*
  • Tryptophan / metabolism
  • Tyrosine / metabolism


  • Homeodomain Proteins
  • Nanog Homeobox Protein
  • Nanog protein, mouse
  • SOXB1 Transcription Factors
  • Sox2 protein, mouse
  • Tyrosine
  • Tryptophan