5-Formylcytosine to Cytosine Conversion by C-C Bond Cleavage in vivo

Iwan, Katharina ; Rahimoff, René ; Kirchner, Angie ; Spada, Fabio ; Schröder, Arne S.; Kosmatchev, Olesea; Ferizaj, Shqiponja; Steinbacher, Jessica; Parsa, Edris; Müller, Markus and Carell, Thomas (27. November 2017): 5-Formylcytosine to Cytosine Conversion by C-C Bond Cleavage in vivo. In: Nature Chemical Biology, Vol. 14: pp. 72-78

Abstract

Tet enzymes oxidise 5-methyl-deoxycytidine (mdC) to 5-hydroxymethyl-dC (hmdC), 5-formyl-dC (fdC) and 5-carboxy-dC (cadC) in DNA. It was proposed that fdC and cadC deformylate and decarboxylate to dC in the course of an active demethylation process. This would re-install canonical dC bases at previously methylated sites. The question whether such direct C-C bond cleavage reactions at fdC and cadC occur in vivo remains an unsolved problem. Here we report the incorporation of synthetic isotope- and (R)-2’-fluorine-labelled dC and fdC-derivatives into the genome of cultured mammalian cells. Following the fate of these probe molecules using UHPLC-MS/MS provided quantitative data about the formed reaction products. The data show that the labelled fdC probe is efficiently converted into the corresponding labelled dC, most likely after its incorporation into the genome. This allows concluding that fdC is undergoing C-C bond cleavage in stem cells that leads to the direct re-installation of unmodified dC.

• Item Type: Journal article
• EU Funded Grant Agreement Number: 741912
• EU Projects: Horizon 2020 > ERC Grants > ERC Advanced Grant > ERC Grant 741912: EPiR - The Chemical Basis of RNA Epigenetics
• Form of publication: Postprint
• Faculties: Chemistry and Pharmacy > Department of Chemistry
• Research Centers: Center for Integrated Protein Science Munich (CIPSM)
• Subjects: 500 Science > 540 Chemistry; 500 Science > 570 Life sciences; biology
• ISSN: 1552-4450
• Language: English