Analysis of the SWI/SNF chromatin-remodeling complex during early heart development and BAF250a repression cardiac gene transcription during P19 cell differentiation

Nucleic Acids Res. 2014 Mar;42(5):2958-75. doi: 10.1093/nar/gkt1232. Epub 2013 Dec 13.

Abstract

The regulatory networks of differentiation programs and the molecular mechanisms of lineage-specific gene regulation in mammalian embryos remain only partially defined. We document differential expression and temporal switching of BRG1-associated factor (BAF) subunits, core pluripotency factors and cardiac-specific genes during post-implantation development and subsequent early organogenesis. Using affinity purification of BRG1 ATPase coupled to mass spectrometry, we characterized the cardiac-enriched remodeling complexes present in E8.5 mouse embryos. The relative abundance and combinatorial assembly of the BAF subunits provides functional specificity to Switch/Sucrose NonFermentable (SWI/SNF) complexes resulting in a unique gene expression profile in the developing heart. Remarkably, the specific depletion of the BAF250a subunit demonstrated differential effects on cardiac-specific gene expression and resulted in arrhythmic contracting cardiomyocytes in vitro. Indeed, the BAF250a physically interacts and functionally cooperates with Nucleosome Remodeling and Histone Deacetylase (NURD) complex subunits to repressively regulate chromatin structure of the cardiac genes by switching open and poised chromatin marks associated with active and repressed gene expression. Finally, BAF250a expression modulates BRG1 occupancy at the loci of cardiac genes regulatory regions in P19 cell differentiation. These findings reveal specialized and novel cardiac-enriched SWI/SNF chromatin-remodeling complexes, which are required for heart formation and critical for cardiac gene expression regulation at the early stages of heart development.

Publication types

  • Research Support, N.I.H., Intramural

MeSH terms

  • Animals
  • Cell Differentiation
  • Cell Line
  • Cell Lineage / genetics
  • Chromatin / metabolism
  • Chromatin Assembly and Disassembly
  • DNA Helicases / metabolism
  • DNA-Binding Proteins / metabolism*
  • Gene Expression Regulation*
  • Heart / embryology*
  • Mi-2 Nucleosome Remodeling and Deacetylase Complex / metabolism
  • Mice
  • Myocardium / cytology
  • Myocardium / metabolism*
  • Nuclear Proteins / metabolism*
  • Protein Subunits / genetics
  • Protein Subunits / metabolism
  • RNA Polymerase II / metabolism
  • Repressor Proteins / metabolism*
  • Transcription Factors / genetics
  • Transcription Factors / metabolism*
  • Transcription, Genetic*

Substances

  • Arid1a protein, mouse
  • Chromatin
  • DNA-Binding Proteins
  • Nuclear Proteins
  • Protein Subunits
  • Repressor Proteins
  • Transcription Factors
  • RNA Polymerase II
  • Mi-2 Nucleosome Remodeling and Deacetylase Complex
  • Smarca4 protein, mouse
  • DNA Helicases