Phosphorylation of the retinoic acid receptor RARγ2 is crucial for the neuronal differentiation of mouse embryonic stem cells

J Cell Sci. 2014 May 1;127(Pt 9):2095-105. doi: 10.1242/jcs.145979. Epub 2014 Feb 25.

Abstract

Retinoic acid (RA) plays key roles in cell differentiation and growth arrest by activating nuclear RA receptors (RARs) (α, β and γ), which are ligand-dependent transcription factors. RARs are also phosphorylated in response to RA. Here, we investigated the in vivo relevance of the phosphorylation of RARs during RA-induced neuronal differentiation of mouse embryonic stem cells (mESCs). Using ESCs where the genes encoding each RAR subtype had been inactivated, and stable rescue lines expressing RARs mutated in phospho-acceptor sites, we show that RA-induced neuronal differentiation involves RARγ2 and requires RARγ2 phosphorylation. By gene expression profiling, we found that the phosphorylated form of RARγ2 regulates a small subset of genes through binding an unusual RA response element consisting of two direct repeats with a seven-base-pair spacer. These new findings suggest an important role for RARγ phosphorylation during cell differentiation and pave the way for further investigations during embryonic development.

Keywords: Differentiation; Nuclear receptor; Phosphorylation; RA; RAR; Retinoic acid; Transcription.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Differentiation / drug effects
  • Embryonic Stem Cells / cytology
  • Embryonic Stem Cells / metabolism
  • Mice
  • Phosphorylation
  • Receptors, Retinoic Acid / metabolism*
  • Tretinoin / pharmacology*

Substances

  • Receptors, Retinoic Acid
  • retinoic acid receptor gamma
  • Tretinoin