Destabilization of the TWIST1/E12 complex dimerization following the R154P point-mutation of TWIST1: an in silico approach

BMC Struct Biol. 2017 May 18;17(1):6. doi: 10.1186/s12900-017-0076-x.

Abstract

Background: The bHLH transcription factor TWIST1 plays a key role in the embryonic development and in tumorigenesis. Some loss-of-function mutations of the TWIST1 gene have been shown to cause an autosomal dominant craniosynostosis, known as the Saethre-Chotzen syndrome (SCS). Although the functional impacts of many TWIST1 mutations have been experimentally reported, little is known on the molecular mechanisms underlying their loss-of-function. In a previous study, we highlighted the predictive value of in silico molecular dynamics (MD) simulations in deciphering the molecular function of TWIST1 residues.

Results: Here, since the substitution of the arginine 154 amino acid by a glycine residue (R154G) is responsible for the SCS phenotype and the substitution of arginine 154 by a proline experimentally decreases the dimerizing ability of TWIST1, we investigated the molecular impact of this point mutation using MD approaches. Consistently, MD simulations highlighted a clear decrease in the stability of the α-helix during the dimerization of the mutated R154P TWIST1/E12 dimer compared to the wild-type TE complex, which was further confirmed in vitro using immunoassays.

Conclusions: Our study demonstrates that MD simulations provide a structural explanation for the loss-of-function associated with the SCS TWIST1 mutation and provides a proof of concept of the predictive value of these MD simulations. This in silico methodology could be used to determine reliable pharmacophore sites, leading to the application of docking approaches in order to identify specific inhibitors of TWIST1 complexes.

Keywords: Dimerization; Molecular dynamics; TWIST1/E12; bHLH transcription factor; in silico analysis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Arginine / chemistry
  • Arginine / genetics
  • Computer Simulation
  • Crystallography, X-Ray
  • Glycine / chemistry
  • Glycine / genetics
  • Humans
  • Mice
  • Molecular Dynamics Simulation*
  • Nuclear Proteins / chemistry*
  • Nuclear Proteins / genetics*
  • Phosphorylation
  • Point Mutation*
  • Protein Conformation
  • Protein Multimerization
  • Sequence Homology
  • Transcription Factor 3 / chemistry*
  • Transcription Factor 3 / genetics
  • Twist-Related Protein 1 / chemistry*
  • Twist-Related Protein 1 / genetics*

Substances

  • Nuclear Proteins
  • TWIST1 protein, human
  • Transcription Factor 3
  • Twist-Related Protein 1
  • Arginine
  • Glycine