High-throughput mapping of a dynamic signaling network in mammalian cells

Science. 2005 Mar 11;307(5715):1621-5. doi: 10.1126/science.1105776.

Abstract

Signaling pathways transmit information through protein interaction networks that are dynamically regulated by complex extracellular cues. We developed LUMIER (for luminescence-based mammalian interactome mapping), an automated high-throughput technology, to map protein-protein interaction networks systematically in mammalian cells and applied it to the transforming growth factor-beta (TGFbeta) pathway. Analysis using self-organizing maps and k-means clustering identified links of the TGFbeta pathway to the p21-activated kinase (PAK) network, to the polarity complex, and to Occludin, a structural component of tight junctions. We show that Occludin regulates TGFbeta type I receptor localization for efficient TGFbeta-dependent dissolution of tight junctions during epithelial-to-mesenchymal transitions.

Publication types

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

MeSH terms

  • Activin Receptors, Type I / metabolism
  • Animals
  • Cell Line
  • Cell Polarity
  • DNA-Binding Proteins / metabolism
  • Epithelial Cells / cytology
  • Epithelial Cells / physiology
  • Humans
  • Immunoprecipitation
  • Luciferases
  • Membrane Proteins / metabolism
  • Mesoderm / cytology
  • Mice
  • Occludin
  • Phosphorylation
  • Protein Interaction Mapping*
  • Protein Serine-Threonine Kinases / metabolism
  • Receptor, Transforming Growth Factor-beta Type I
  • Receptor, Transforming Growth Factor-beta Type II
  • Receptors, Transforming Growth Factor beta / metabolism
  • Recombinant Fusion Proteins / metabolism
  • Signal Transduction*
  • Smad2 Protein
  • Smad4 Protein
  • Tight Junctions / ultrastructure
  • Trans-Activators / metabolism
  • Transforming Growth Factor beta / metabolism*
  • p21-Activated Kinases

Substances

  • DNA-Binding Proteins
  • Membrane Proteins
  • OCLN protein, human
  • Occludin
  • Ocln protein, mouse
  • Receptors, Transforming Growth Factor beta
  • Recombinant Fusion Proteins
  • SMAD2 protein, human
  • SMAD4 protein, human
  • Smad2 Protein
  • Smad2 protein, mouse
  • Smad4 Protein
  • Trans-Activators
  • Transforming Growth Factor beta
  • Luciferases
  • PAK1 protein, human
  • Pak1 protein, mouse
  • Protein Serine-Threonine Kinases
  • p21-Activated Kinases
  • Activin Receptors, Type I
  • Receptor, Transforming Growth Factor-beta Type I
  • Receptor, Transforming Growth Factor-beta Type II