TY - JOUR
T1 - TGF-β uses a novel mode of receptor activation to phosphorylate SMAD1/5 and induce epithelial-to-mesenchymal transition
AU - Ramachandran, Anassuya
AU - Vizán, Pedro
AU - Das, Debipriya
AU - Chakravarty, Probir
AU - Vogt, Janis
AU - Rogers, Katherine W.
AU - Müller, Patrick
AU - Hinck, Andrew P.
AU - Sapkota, Gopal P.
AU - Hill, Caroline S.
N1 - Funding Information:
We thank Lalage Wakefield for providing the TGF-b neutralizing antibody and the isotype-matched control, Paul Yu for LDN-193189, Christian Bökel for the pCS2+zALK3 IPF expression plasmid and Bob Lechleider for the FLAG-SMAD1 expression plasmid. We thank Nik Matthews, Greg Elgar and the Advanced Sequencing Facility for the next generation sequencing. We are grateful to the Francis Crick Institute Light Microscopy and Flow Cytometry facilities and to the Genomics Equipment Park. We thank Alex Bullock for very fruitful discussions and all the members of the Hill lab for useful comments on the manuscript. This work was supported by the Francis Crick Institute which receives its core funding from Cancer Research UK (FC001095), the UK Medical Research Council (FC001095), and the Wellcome Trust (FC001095). The development and characterization of TGF-b3WD in the Hinck laboratory was enabled by support provided by the NIH (GM58670 and CA172886).
Funding Information:
We thank Lalage Wakefield for providing the TGF-β neutralizing antibody and the isotype-matched control, Paul Yu for LDN-193189, Christian Bökel for the pCS2+zALK3 IPF expression plasmid and Bob Lechleider for the FLAG-SMAD1 expression plasmid. We thank Nik Matthews, Greg Elgar and the Advanced Sequencing Facility for the next generation sequencing. We are grateful to the Francis Crick Institute Light Microscopy and Flow Cytometry facilities and to the Genomics Equipment Park. We thank Alex Bullock for very fruitful discussions and all the members of the Hill lab for useful comments on the manuscript. This work was supported by the Francis Crick Institute which receives its core funding from Cancer Research UK (FC001095), the UK Medical Research Council (FC001095), and the Wellcome Trust (FC001095). The development and characterization of TGF-β3WD in the Hinck laboratory was enabled by support provided by the NIH (GM58670 and CA172886). Francis Crick Institute FC001095 Anassuya Ramachandran Pedro Vizan Debipriya Das Probir Chakravarty Caroline S Hill NIH Office of the Director GM58670 Andrew P Hinck NIH Office of the Director CA172886 Andrew P Hinck The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Publisher Copyright:
© Ramachandran et al.
PY - 2018/1/29
Y1 - 2018/1/29
N2 - The best characterized signaling pathway downstream of transforming growth factor b (TGF-β) is through SMAD2 and SMAD3. However, TGF-β also induces phosphorylation of SMAD1 and SMAD5, but the mechanism of this phosphorylation and its functional relevance is not known. Here, we show that TGF-b-induced SMAD1/5 phosphorylation requires members of two classes of type I receptor, TGFBR1 and ACVR1, and establish a new paradigm for receptor activation where TGFBR1 phosphorylates and activates ACVR1, which phosphorylates SMAD1/5. We demonstrate the biological significance of this pathway by showing that approximately a quarter of the TGF-β- induced transcriptome depends on SMAD1/5 signaling, with major early transcriptional targets being the ID genes. Finally, we show that TGF-b-induced epithelial-to-mesenchymal transition requires signaling via both the SMAD3 and SMAD1/5 pathways, with SMAD1/5 signaling being essential to induce ID1. Therefore, combinatorial signaling via both SMAD pathways is essential for the full TGF-β-induced transcriptional program and physiological responses.
AB - The best characterized signaling pathway downstream of transforming growth factor b (TGF-β) is through SMAD2 and SMAD3. However, TGF-β also induces phosphorylation of SMAD1 and SMAD5, but the mechanism of this phosphorylation and its functional relevance is not known. Here, we show that TGF-b-induced SMAD1/5 phosphorylation requires members of two classes of type I receptor, TGFBR1 and ACVR1, and establish a new paradigm for receptor activation where TGFBR1 phosphorylates and activates ACVR1, which phosphorylates SMAD1/5. We demonstrate the biological significance of this pathway by showing that approximately a quarter of the TGF-β- induced transcriptome depends on SMAD1/5 signaling, with major early transcriptional targets being the ID genes. Finally, we show that TGF-b-induced epithelial-to-mesenchymal transition requires signaling via both the SMAD3 and SMAD1/5 pathways, with SMAD1/5 signaling being essential to induce ID1. Therefore, combinatorial signaling via both SMAD pathways is essential for the full TGF-β-induced transcriptional program and physiological responses.
UR - https://www.scopus.com/pages/publications/85043513491
U2 - 10.7554/eLife.31756
DO - 10.7554/eLife.31756
M3 - Article
C2 - 29376829
AN - SCOPUS:85043513491
SN - 2050-084X
VL - 7
JO - eLife
JF - eLife
M1 - e31756
ER -