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Functional role of Tet-mediated RNA hydroxymethylcytosine in mouse ES cells and during differentiation.


Lan, Jie and Rajan, Nicholas and Bizet, Martin and Penning, Audrey and Singh, Nitesh K and Guallar, Diana and Calonne, Emilie and Li Greci, Andrea and Bonvin, Elise and Deplus, Rachel and Hsu, Phillip J and Nachtergaele, Sigrid and Ma, Chengjie and Song,

Tet-enzyme-mediated 5-hydroxymethylation of cytosines in DNA plays a crucial role in mouse embryonic stem cells (ESCs). In RNA also, 5-hydroxymethylcytosine (5hmC) has recently been evidenced, but its physiological roles are still largely unknown. Here we show the contribution and function of this mark in mouse ESCs and differentiating embryoid bodies. Transcriptome-wide mapping in ESCs reveals hundreds of messenger RNAs marked by 5hmC at sites characterized by a defined unique consensus sequence and particular features. During differentiation a large number of transcripts, including many encoding key pluripotency-related factors (such as Eed and Jarid2), show decreased cytosine hydroxymethylation. Using Tet-knockout ESCs, we find Tet enzymes to be partly responsible for deposition of 5hmC in mRNA. A transcriptome-wide search further reveals mRNA targets to which Tet1 and Tet2 bind, at sites showing a topology similar to that of 5hmC sites. Tet-mediated RNA hydroxymethylation is found to reduce the stability of crucial pluripotency-promoting transcripts. We propose that RNA cytosine 5-hydroxymethylation by Tets is a mark of transcriptome flexibility, inextricably linked to the balance between pluripotency and lineage commitment.

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Published
October, 2020

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Products used in this publication

  • Mouse IgG
    C15220001-50
    5-hydroxymethylcytosine (5-hmC) monoclonal anti...

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