High-throughput mapping of a dynamic signaling network in mammalian cells.
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.
Barrios-Rodiles
Miriam
M
Program in Molecular Biology and Cancer, Samuel Lunenfeld Research Institute, Mount Sinai Hospital, Toronto, Ontario, Canada, M5G 1X5.
Brown
Kevin R
KR
Ozdamar
Barish
B
Bose
Rohit
R
Liu
Zhong
Z
Donovan
Robert S
RS
Shinjo
Fukiko
F
Liu
Yongmei
Y
Dembowy
Joanna
J
Taylor
Ian W
IW
Luga
Valbona
V
Przulj
Natasa
N
Robinson
Mark
M
Suzuki
Harukazu
H
Hayashizaki
Yoshihide
Y
Jurisica
Igor
I
Wrana
Jeffrey L
JL
D030201:Activin Receptors, Type I / Q000378:metabolism; D000818:Animals; D002460:Cell Line; D016764:Cell Polarity; D004268:DNA-Binding Proteins / Q000378:metabolism; D004847:Epithelial Cells / Q000166:cytology / Q000502:physiology; D006801:Humans; D047468:Immunoprecipitation; D008156:Luciferases; D008565:Membrane Proteins / Q000378:metabolism; D008648:Mesoderm / Q000166:cytology; D051379:Mice; D062793:Occludin; D010766:Phosphorylation; D025941:Protein Interaction Mapping; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism; D000077293:Receptor, Transforming Growth Factor-beta Type I; D000077294:Receptor, Transforming Growth Factor-beta Type II; D018125:Receptors, Transforming Growth Factor beta / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D015398:Signal Transduction; D051899:Smad2 Protein; D051901:Smad4 Protein; D019108:Tight Junctions / Q000648:ultrastructure; D015534:Trans-Activators / Q000378:metabolism; D016212:Transforming Growth Factor beta / Q000378:metabolism*; D054462:p21-Activated Kinases
D004268:DNA-Binding Proteins; D008565:Membrane Proteins; C568985:OCLN protein, human; D062793:Occludin; C568986:Ocln protein, mouse; D018125:Receptors, Transforming Growth Factor beta; D011993:Recombinant Fusion Proteins; C497782:SMAD2 protein, human; C497796:SMAD4 protein, human; D051899:Smad2 Protein; C497783:Smad2 protein, mouse; D051901:Smad4 Protein; D015534:Trans-Activators; D016212:Transforming Growth Factor beta; D008156:Luciferases; C515903:PAK1 protein, human; C515905:Pak1 protein, mouse; D017346:Protein Serine-Threonine Kinases; D054462:p21-Activated Kinases; D030201:Activin Receptors, Type I; D000077293:Receptor, Transforming Growth Factor-beta Type I; D000077294:Receptor, Transforming Growth Factor-beta Type II