Interaction between Smad 3 and Dishevelled in murine embryonic craniofacial mesenchymal cells.
OBJECTIVE
To determine the in vivo interaction between Smad 3 and Dishevelled-1.


METHODS
Cell culture transfection followed by immunoprecipitation with specific antibodies.


METHODS
The Department of Molecular, Cellular, and Craniofacial Biology, Birth Defects Center, University of Louisville.


METHODS
Overexpression of myc-Smad 3.


METHODS
Western blotting of anti-Dishevelled immunoprecipitates for Smad 3.


RESULTS
Smad 3 and Dishevelled isoforms-1, -2, and -3 all bind Smad 3 in glutathione-S-transferase (GST) pull-down assays and Smad 3 binds to Dishevelled-1 in vivo. Stimulation of the transforming growth factor beta (TGFbeta) pathway leads to increased binding of Smad 3 and Dishevelled-1 in vivo.


CONCLUSIONS
Smad 3 binds all three known isoforms of Dishevelled and binds Dishevelled 1 in vivo. TGFbeta signaling modulates the interaction between Smad 3 and Dishevelled-1.
Warner
D R
DR
Department of Molecular, Cellular, and Craniofacial Biology, University of Louisville Birth Defects Center, Louisville, KY 40292, USA.
Greene
R M
RM
Pisano
M M
MM
D048868:Adaptor Proteins, Signal Transducing; D000818:Animals; D002478:Cells, Cultured; D003470:Culture Media; D004268:DNA-Binding Proteins / Q000502:physiology*; D000072261:Dishevelled Proteins; D004622:Embryo, Mammalian; D047108:Embryonic Development / Q000502:physiology; D005982:Glutathione Transferase; D036341:Intercellular Signaling Peptides and Proteins / Q000502:physiology; D008437:Maxilla / Q000166:cytology / Q000196:embryology*; D008648:Mesoderm / Q000166:cytology* / Q000502:physiology; D051379:Mice; D008813:Mice, Inbred ICR; D010159:Palate / Q000166:cytology / Q000196:embryology; D010750:Phosphoproteins; D011485:Protein Binding / Q000502:physiology; D011505:Protein-Tyrosine Kinases / Q000502:physiology; D011506:Proteins / Q000502:physiology*; D015398:Signal Transduction / Q000502:physiology*; D051900:Smad3 Protein; D015534:Trans-Activators / Q000502:physiology*; D016212:Transforming Growth Factor beta / Q000502:physiology*; D051153:Wnt Proteins
D048868:Adaptor Proteins, Signal Transducing; D003470:Culture Media; D004268:DNA-Binding Proteins; D000072261:Dishevelled Proteins; C000606579:Dvl1 protein, mouse; D036341:Intercellular Signaling Peptides and Proteins; D010750:Phosphoproteins; D011506:Proteins; D051900:Smad3 Protein; C497791:Smad3 protein, mouse; D015534:Trans-Activators; D016212:Transforming Growth Factor beta; D051153:Wnt Proteins; D005982:Glutathione Transferase; D011505:Protein-Tyrosine Kinases