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Oxidative and non-oxidative active turnover of genomic methylcytosine in distinct pluripotent states

By Fabio Spada, Sarah Schiffers, Angie Kirchner, Yingqian Zhang, Olesea Kosmatchev, Eva Korytiakova, René Rahimoff, Charlotte Ebert, Thomas Carell

Posted 18 Nov 2019
bioRxiv DOI: 10.1101/846584

Epigenetic plasticity underpins cell potency, but the extent to which active turnover of DNA methylation contributes to such plasticity is not known and the underlying pathways are poorly understood. Here we use mass spectrometry in combination with isoptope tracing to quantitatively address the global turnover of genomic methylcytidine (mdC), hydroxymethylcytidine (hmdC) and formylcytidine (fdC) across mouse pluripotent cell states. High rates of mdC/hmdC oxidation and fdC turnover characterize a formative-like pluripotent state. In primed pluripotent cells the global mdC turnover rate is about 3-6% faster than can be explained by passive dilution through DNA synthesis. While this active component is largely dependent on Tet-mediated mdC oxidation, we also unveiled an additional mdC oxidation-independent turnover process based on DNA repair. This process accelerates upon acquisition of primed pluripotency and returns to low levels in lineage committed cells. Thus, in pluripotent cells active mdC turnover involves both mdC oxidation-dependent and independent processes.

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