title: A direct physical interaction between Nanog and Sox2 regulates embryonic stem cell self-renewal.
abstract: 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.
authors.affiliation: MRC Centre for Regenerative Medicine, Institute for Stem Cell Research, School of Biological Sciences, University of Edinburgh, Edinburgh, UK.
authors.forename: Alessia
authors.initials: A
authors.lastname: Gagliardi
authors.forename: Nicholas P
authors.initials: NP
authors.lastname: Mullin
authors.forename: Zi
authors.initials: Z
authors.lastname: Ying Tan
authors.forename: Douglas
authors.initials: D
authors.lastname: Colby
authors.forename: Anastasia I
authors.initials: AI
authors.lastname: Kousa
authors.forename: Florian
authors.initials: F
authors.lastname: Halbritter
authors.forename: Jason T
authors.initials: JT
authors.lastname: Weiss
authors.forename: Anastasia
authors.initials: A
authors.lastname: Felker
authors.forename: Karel
authors.initials: K
authors.lastname: Bezstarosti
authors.forename: Rebecca
authors.initials: R
authors.lastname: Favaro
authors.forename: Jeroen
authors.initials: J
authors.lastname: Demmers
authors.forename: Silvia K
authors.initials: SK
authors.lastname: Nicolis
authors.forename: Simon R
authors.initials: SR
authors.lastname: Tomlinson
authors.forename: Raymond A
authors.initials: RA
authors.lastname: Poot
authors.forename: Ian
authors.identifier: 0000000326051597
authors.initials: I
authors.lastname: Chambers
mesh_terms: D000818:Animals; D049109:Cell Proliferation; D003114:Colony-Forming Units Assay; D053595:Embryonic Stem Cells / Q000378:metabolism / Q000502:physiology*; D018398:Homeodomain Proteins / Q000378:metabolism*; D015151:Immunoblotting; D047468:Immunoprecipitation; D051379:Mice; D000071317:Nanog Homeobox Protein; D010957:Plasmids / Q000235:genetics; D054730:Protein Interaction Domains and Motifs / Q000235:genetics*; D025941:Protein Interaction Mapping; D052156:SELEX Aptamer Technique; D055748:SOXB1 Transcription Factors / Q000378:metabolism*; D014364:Tryptophan / Q000378:metabolism; D014443:Tyrosine / Q000378:metabolism
chemical_list: D018398:Homeodomain Proteins; D000071317:Nanog Homeobox Protein; C475540:Nanog protein, mouse; D055748:SOXB1 Transcription Factors; C527783:Sox2 protein, mouse; D014443:Tyrosine; D014364:Tryptophan
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