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A PRC2-dependent repressive role of PRDM14 in human embryonic stem cells and induced pluripotent stem cell

Yun-Shen Chan1, Jonathan Göke, Xinyi Lu

  • 1Gene Regulation Laboratory, Genome Institute of Singapore, Singapore, Singapore.

Stem Cells (Dayton, Ohio)
|January 3, 2013
PubMed

Insights

PRDM14 directly represses developmental genes in human stem cells by recruiting PRC2, a chromatin regulator. This mechanism is crucial for maintaining pluripotency and reprogramming cells, revealing PRDM14 as a new PRC2 regulator.

Area of Science:

  • Stem cell biology
  • Epigenetics
  • Gene regulation

Background:

  • PRDM14 is key for human embryonic stem cell (ESC) identity and pluripotency.
  • PRDM14's repressive mechanism in human ESCs was previously unknown.
  • While known to repress genes in mouse cells, its function in human cells needed clarification.

Purpose of the Study:

  • To elucidate the mechanism of PRDM14's repressive activity in human ESCs.
  • To investigate PRDM14's role in the maintenance and induction of pluripotency.
  • To identify PRDM14 as a regulator of chromatin modifiers.

Main Methods:

  • Chromatin immunoprecipitation sequencing (ChIP-seq) to map PRDM14 binding sites.
  • Analysis of histone modifications, specifically H3K27me3.
  • Co-immunoprecipitation assays to study PRDM14-PRC2 interaction.
  • Reporter assays to assess gene repression.
  • Experiments involving PRDM14 depletion and ectopic expression.
  • Analysis of ZEB1 repression during induced pluripotent stem cell (iPSC) reprogramming.

Main Results:

  • PRDM14 directly represses developmental genes in human ESCs by binding to silenced loci.
  • PRDM14 binding sites are enriched for H3K27me3, a repressive histone mark.
  • PRDM14 directly interacts with Polycomb Repressive Complex 2 (PRC2).
  • PRDM14 recruits PRC2 to target loci, leading to H3K27me3 deposition and gene repression.
  • PRDM14 depletion reduces PRC2 binding and H3K27me3 levels.
  • PRDM14-mediated repression is dependent on both PRDM14 and PRC2.
  • Ectopic PRDM14 represses developmental genes in fibroblasts and recruits PRC2 to repress ZEB1, facilitating mesenchymal-to-epithelial transition during iPSC reprogramming.

Conclusions:

  • PRDM14 plays a direct repressive role in maintaining pluripotency in human ESCs.
  • PRDM14 is a novel regulator that recruits PRC2 to mediate gene silencing.
  • This mechanism is important for both stem cell identity and the reprogramming of somatic cells into iPSCs.

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