Structural basis for genome wide recognition of 5-bp GC motifs by SMAD transcription factors

Nat Commun. 2017 Dec 12;8(1):2070. doi: 10.1038/s41467-017-02054-6.

Abstract

Smad transcription factors activated by TGF-β or by BMP receptors form trimeric complexes with Smad4 to target specific genes for cell fate regulation. The CAGAC motif has been considered as the main binding element for Smad2/3/4, whereas Smad1/5/8 have been thought to preferentially bind GC-rich elements. However, chromatin immunoprecipitation analysis in embryonic stem cells showed extensive binding of Smad2/3/4 to GC-rich cis-regulatory elements. Here, we present the structural basis for specific binding of Smad3 and Smad4 to GC-rich motifs in the goosecoid promoter, a nodal-regulated differentiation gene. The structures revealed a 5-bp consensus sequence GGC(GC)|(CG) as the binding site for both TGF-β and BMP-activated Smads and for Smad4. These 5GC motifs are highly represented as clusters in Smad-bound regions genome-wide. Our results provide a basis for understanding the functional adaptability of Smads in different cellular contexts, and their dependence on lineage-determining transcription factors to target specific genes in TGF-β and BMP pathways.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Allosteric Regulation / genetics
  • Amino Acid Motifs*
  • Animals
  • Binding Sites / genetics
  • Bone Morphogenetic Proteins / metabolism
  • CRISPR-Cas Systems
  • Cell Lineage / genetics
  • Crystallography, X-Ray
  • Gene Expression Regulation, Developmental
  • Goosecoid Protein / genetics*
  • Magnetic Resonance Spectroscopy
  • Mice
  • Mouse Embryonic Stem Cells
  • Promoter Regions, Genetic
  • Protein Binding
  • Smad3 Protein / chemistry*
  • Smad3 Protein / genetics
  • Smad4 Protein / chemistry*
  • Smad4 Protein / genetics
  • Transforming Growth Factor beta / metabolism

Substances

  • Bone Morphogenetic Proteins
  • GSC protein, human
  • Goosecoid Protein
  • SMAD3 protein, human
  • SMAD4 protein, human
  • Smad3 Protein
  • Smad4 Protein
  • Transforming Growth Factor beta