Directed network wiring identifies a key protein interaction in embryonic stem cell differentiation

Mol Cell. 2014 Jun 19;54(6):1034-41. doi: 10.1016/j.molcel.2014.05.002. Epub 2014 Jun 5.

Abstract

Cell signaling depends on dynamic protein-protein interaction (PPI) networks, often assembled through modular domains each interacting with multiple peptide motifs. This complexity raises a conceptual challenge, namely to define whether a particular cellular response requires assembly of the complete PPI network of interest or can be driven by a specific interaction. To address this issue, we designed variants of the Grb2 SH2 domain ("pY-clamps") whose specificity is highly biased toward a single phosphotyrosine (pY) motif among many potential pYXNX Grb2-binding sites. Surprisingly, directing Grb2 predominantly to a single pY site of the Ptpn11/Shp2 phosphatase, but not other sites tested, was sufficient for differentiation of the essential primitive endoderm lineage from embryonic stem cells. Our data suggest that discrete connections within complex PPI networks can underpin regulation of particular biological events. We propose that this directed wiring approach will be of general utility in functionally annotating specific PPIs.

Publication types

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

MeSH terms

  • Animals
  • Binding Sites / genetics
  • Cell Differentiation* / genetics
  • Cell Line
  • Crystallography, X-Ray
  • Embryonic Stem Cells / cytology*
  • Embryonic Stem Cells / metabolism
  • Fibroblast Growth Factor 4 / metabolism
  • GRB2 Adaptor Protein / genetics
  • GRB2 Adaptor Protein / metabolism*
  • Mice
  • Models, Molecular
  • Protein Binding / genetics
  • Protein Interaction Maps / genetics*
  • Protein Structure, Tertiary
  • Protein Tyrosine Phosphatase, Non-Receptor Type 11 / genetics
  • Protein Tyrosine Phosphatase, Non-Receptor Type 11 / metabolism*
  • Protein Tyrosine Phosphatase, Non-Receptor Type 11 / ultrastructure
  • Signal Transduction / genetics

Substances

  • Fgf4 protein, mouse
  • Fibroblast Growth Factor 4
  • GRB2 Adaptor Protein
  • Grb2 protein, mouse
  • Protein Tyrosine Phosphatase, Non-Receptor Type 11
  • Ptpn11 protein, mouse

Associated data

  • PDB/4JMG
  • PDB/4JMH