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Visualization and Quantification of TGFβ/BMP/SMAD Signaling under Different Fluid Shear Stress Conditions using Proximity-Ligation-Assay
Published on: September 14, 2021
The tumor suppressor Smad4/DPC4 is regulated by phosphorylations that integrate FGF, Wnt, and TGF-β signaling
Hadrien Demagny1, Tatsuya Araki1, Edward M De Robertis1
1Howard Hughes Medical Institute and Department of Biological Chemistry, University of California, Los Angeles, Los Angeles, CA 90095-1662, USA.
Abstract:
Smad4 is a major tumor suppressor currently thought to function constitutively in the transforming growth factor β (TGF-β)-signaling pathway. Here, we report that Smad4 activity is directly regulated by the Wnt and fibroblast growth factor (FGF) pathways through GSK3 and mitogen-activated protein kinase (MAPK) phosphorylation sites. FGF activates MAPK, which primes three sequential GSK3 phosphorylations that generate a Wnt-regulated phosphodegron bound by the ubiquitin E3 ligase β-TrCP. In the presence of FGF, Wnt potentiates TGF-β signaling by preventing Smad4 GSK3 phosphorylations that inhibit a transcriptional activation domain located in the linker region. When MAPK is not activated, the Wnt and TGF-β signaling pathways remain insulated from each other. In Xenopus embryos, these Smad4 phosphorylations regulate germ-layer specification and Spemann organizer formation. The results show that three major signaling pathways critical in development and cancer are integrated at the level of Smad4.
Insights
Smad4 tumor suppressor activity is regulated by Wnt and FGF signaling pathways via phosphorylation. This integration impacts key developmental processes and cancer, revealing cross-talk between major signaling cascades.
Area of Science:
- Molecular Biology
- Cell Signaling
- Developmental Biology
Background:
- Smad4 is a key tumor suppressor in the transforming growth factor β (TGF-β) pathway.
- Its constitutive function has been the prevailing model in cancer research.
Purpose of the Study:
- To investigate the regulatory mechanisms of Smad4 activity beyond the canonical TGF-β pathway.
- To elucidate how Wnt and fibroblast growth factor (FGF) signaling pathways intersect with Smad4 function.
Main Methods:
- Phosphorylation site analysis of Smad4.
- Investigating the roles of glycogen synthase kinase 3 (GSK3) and mitogen-activated protein kinase (MAPK).
- Utilizing Xenopus embryo models to study developmental implications.
Main Results:
- Smad4 activity is modulated by Wnt and FGF pathways through specific phosphorylation sites.
- FGF-activated MAPK primes Smad4 for Wnt-dependent ubiquitination via GSK3 and β-TrCP.
- This cross-talk regulates TGF-β signaling, germ-layer specification, and Spemann organizer formation in Xenopus.
Conclusions:
- Smad4 integrates Wnt, FGF, and TGF-β signaling pathways through phosphorylation-dependent mechanisms.
- This integration is crucial for normal development and has implications for understanding cancer.
- The findings challenge the notion of constitutive Smad4 function, highlighting dynamic regulation.
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