# REST missense mutations reveal disrupted Re1 motif binding and co-repressor interactions in uterine fibroids

## TL;DR

This study shows how mutations in the REST protein disrupt its ability to control gene activity, potentially contributing to the development of uterine fibroids.

## Contribution

The study identifies specific REST mutations that impair co-repressor and DNA interactions, offering new insights into REST's role in uterine fibroid biology.

## Key findings

- Five REST variants (Y31C, Y31D, L76Q, Y283C, L427Q) were identified as structurally disruptive.
- The Y283C mutation significantly reduced REST's interaction with SIN3A and DNA binding.
- REST destabilization through Y283C may lead to transcriptional dysregulation and fibroid growth.

## Abstract

The Re1-Silencing Transcription Factor (REST) is a master regulator of gene silencing, orchestrating transcriptional repression by tethering chromatin-modifying co-repressors to the Re1 motif of target genes. While REST is recognized as a sentinel of cellular identity, its role in uterine fibroids (UF) remains unclear. This study aims to investigate how structural perturbations in REST may compromise its regulatory function and contribute to altered transcriptional control in fibroid biology.

A deep structural interrogation of REST was performed through expansive in silico analysis of 938 missense SNPs. Evolutionary conservation was assessed across ten primate species to identify structurally disruptive variants. Structural modelling, protein–protein and protein–DNA docking analyses were conducted to evaluate interactions with co-repressors and DNA. Molecular dynamics simulations were used to assess conformational stability, flexibility, compactness, and energetic changes in wild-type and mutant REST variants.

Five structurally disruptive REST variants (Y31C, Y31D, L76Q, Y283C, L427Q) were identified at evolutionarily conserved residues. Structural modelling and docking analyses revealed weakened affinity for co-repressors, with the Y283C variant showing a marked reduction in SIN3A interaction (Z-score: 2.4 to −1.2) and impaired DNA binding (Z-score: 2.0 to −1.3). Molecular dynamics simulations demonstrated that Y283C increased rigidity (RMSF: 0.33 to 0.27 nm), reduced compactness (Rg: 3.48–3.51 nm), and lowered potential energy. Upon Re1 binding, destabilization intensified, with increased RMSD (0.95–1.07 nm) and pronounced shifts in energy.

This integrative analysis highlights REST as a candidate regulatory component in uterine fibroid biology. Structural disruption of REST, particularly through the Y283C mutation, may destabilize molecular interactions and compromise DNA-binding precision, potentially unleashing transcriptional noise that fuels fibroid growth. These findings suggest that perturbation of REST-mediated transcriptional repression may be associated with altered regulatory control in this disease and could inform future strategies to investigate dysregulation in uterine fibroids.

Investigating the Role of REST Mutational Destabilization in Uterine Leiomyoma Formation via In Silico Docking and Molecular Dynamics Simulations. This graphical abstract depicts the computational workflow exploring the impact of single nucleotide polymorphisms (SNPs) on REST protein stability and its role in uterine leiomyoma formation. Homology modelling of REST was followed by SNP-based mutational induction, with destabilizing mutations predicted through stability analysis. Protein-protein and protein-DNA docking were employed to assess the effects of mutations on REST interactions with key partners and the Re1 motif. Molecular dynamics (MD) simulations were then performed to evaluate the structural destabilization of REST mutants, linking mutational inactivation to GPR10 overexpression, a proposed driver of uterine leiomyoma pathogenesis.Diagram illustrating the role of REST protein in leiomyoma formation. It shows a normal uterus with normal REST protein and a uterus with leiomyoma featuring mutated REST. The process includes REST mutational inactivation, GPR10 overexpression, and REST's identification in leiomyoma formation. The REST protein is modeled, followed by single nucleotide polymorphism (SNP) analysis highlighting mutations. A molecular dynamics simulation and protein analysis, including protein-protein docking and protein-DNA docking, are depicted. A mutational destabilization of the REST protein influences GPR10 overexpression, promoting leiomyoma.

Investigating the Role of REST Mutational Destabilization in Uterine Leiomyoma Formation via In Silico Docking and Molecular Dynamics Simulations. This graphical abstract depicts the computational workflow exploring the impact of single nucleotide polymorphisms (SNPs) on REST protein stability and its role in uterine leiomyoma formation. Homology modelling of REST was followed by SNP-based mutational induction, with destabilizing mutations predicted through stability analysis. Protein-protein and protein-DNA docking were employed to assess the effects of mutations on REST interactions with key partners and the Re1 motif. Molecular dynamics (MD) simulations were then performed to evaluate the structural destabilization of REST mutants, linking mutational inactivation to GPR10 overexpression, a proposed driver of uterine leiomyoma pathogenesis.