The nucleosome remodeling and deacetylase complex protein CHD4 regulates neural differentiation of mouse embryonic stem cells by down-regulating p53

J Biol Chem. 2019 Jan 4;294(1):195-209. doi: 10.1074/jbc.RA118.004086. Epub 2018 Nov 8.

Abstract

Lineage specification of the three germ layers occurs during early embryogenesis and is critical for normal development. The nucleosome remodeling and deacetylase (NuRD) complex is a repressive chromatin modifier that plays a role in lineage commitment. However, the role of chromodomain helicase DNA-binding protein 4 (CHD4), one of the core subunits of the NuRD complex, in neural lineage commitment is poorly understood. Here, we report that the CHD4/NuRD complex plays a critical role in neural differentiation of mouse embryonic stem cells (ESCs). We found that RNAi-mediated Chd4 knockdown suppresses neural differentiation, as did knockdown of methyl-CpG-binding domain protein Mbd3, another NuRD subunit. Chd4 and Mbd3 knockdowns similarly affected changes in global gene expression during neural differentiation and up-regulated several mesendodermal genes. However, inhibition of mesendodermal genes by knocking out the master regulators of mesendodermal lineages, Brachyury and Eomes, through a CRISPR/Cas9 approach could not restore the impaired neural differentiation caused by the Chd4 knockdown, suggesting that CHD4 controls neural differentiation by not repressing other lineage differentiation processes. Notably, Chd4 knockdown increased the acetylation levels of p53, resulting in increased protein levels of p53. Double knockdown of Chd4 and p53 restored the neural differentiation rate. Furthermore, overexpression of BCL2, a downstream factor of p53, partially rescued the impaired neural differentiation caused by the Chd4 knockdown. Our findings reveal that the CHD4/NuRD complex regulates neural differentiation of ESCs by down-regulating p53.

Keywords: chromodomain helicase DNA-binding protein 4 (CHD4); embryonic stem cell; lineage specification; methyl-CpG binding domain protein 3 (MBD3); neurodifferentiation; nucleosome remodeling deacetylase (NuRD); p53; protein degradation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Differentiation*
  • Cell Line
  • DNA Helicases / genetics
  • DNA Helicases / metabolism*
  • Down-Regulation*
  • Gene Knockdown Techniques
  • Mice
  • Mouse Embryonic Stem Cells
  • Neurons / cytology
  • Neurons / metabolism*
  • Nucleosomes / genetics
  • Nucleosomes / metabolism*
  • Proto-Oncogene Proteins c-bcl-2 / biosynthesis
  • Proto-Oncogene Proteins c-bcl-2 / genetics
  • Tumor Suppressor Protein p53 / biosynthesis*
  • Tumor Suppressor Protein p53 / genetics

Substances

  • Nucleosomes
  • Proto-Oncogene Proteins c-bcl-2
  • Trp53 protein, mouse
  • Tumor Suppressor Protein p53
  • Bcl2 protein, mouse
  • Mi-2beta protein, mouse
  • DNA Helicases