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Immunohistochemical Detection of 5-Methylcytosine and 5-Hydroxymethylcytosine in Developing and Postmitotic Mouse Retina
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Functional role of Tet-mediated RNA hydroxymethylcytosine in mouse ES cells and during differentiation

Jie Lan1, Nicholas Rajan1, Martin Bizet1

  • 1Laboratory of Cancer Epigenetics, Faculty of Medicine, ULB Cancer Research Center (U-CRC), Welbio Investigator, Université Libre de Bruxelles (ULB), Brussels, Belgium.

Nature Communications
|October 3, 2020
PubMed

Insights

Tet enzymes add a 5-hydroxymethylcytosine (5hmC) mark to messenger RNA (mRNA) in mouse stem cells. This RNA modification impacts gene expression and cell differentiation.

Area of Science:

  • Epigenetics
  • Molecular Biology
  • Stem Cell Biology

Background:

  • Tet-enzyme-mediated 5-hydroxymethylation is vital for DNA in mouse embryonic stem cells (ESCs).
  • The role of 5-hydroxymethylcytosine (5hmC) in RNA is not well understood.
  • Investigating 5hmC in RNA is crucial for understanding gene regulation and cell fate.

Purpose of the Study:

  • To map and characterize 5hmC marks on messenger RNA (mRNA) in mouse ESCs.
  • To investigate the function of RNA 5hmC during cell differentiation.
  • To determine the role of Tet enzymes in RNA hydroxymethylation.

Main Methods:

  • Transcriptome-wide mapping of 5hmC in ESCs and embryoid bodies.
  • Analysis of Tet enzyme binding sites on mRNA.
  • Utilizing Tet-knockout ESCs to study enzyme function.

Main Results:

  • Hundreds of mRNAs in ESCs exhibit 5hmC at specific consensus sequences.
  • Differentiation leads to decreased 5hmC on transcripts encoding pluripotency factors.
  • Tet enzymes are involved in depositing 5hmC onto mRNA.
  • Tet1 and Tet2 bind to mRNA targets with similar topology to 5hmC sites.
  • Tet-mediated RNA hydroxymethylation reduces the stability of pluripotency-promoting transcripts.

Conclusions:

  • Tet-mediated RNA 5-hydroxymethylation is a novel epigenetic mark.
  • This modification influences transcriptome flexibility and the balance between pluripotency and differentiation.
  • RNA 5hmC acts as a regulatory mechanism in stem cell fate decisions.

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