Pyrene-based Quantitative Detection of the 5-formylcytosine Loci Symmetry in the CpG Duplex Content During TET-dependent Demethylation

Overview

Journal Angew Chem Int Ed Engl

Specialty Chemistry

Date 2014 Aug 28

PMID 25159856

Citations 14

Authors

Liang Xu

Ying-Chu Chen

Jenny Chong

Andrea Fin

Lisa S McCoy

Jun Xu

Chao Zhang

Dong Wang

Affiliations

Soon will be listed here.

Abstract

Methylcytosine (5mC) is mostly symmetrically distributed in CpG sites. Ten-eleven-translocation (TET) proteins are the key enzymes involved in active DNA demethylation through stepwise oxidation of 5mC. However, oxidation pathways of TET enzymes in the symmetrically methylated CpG context are still elusive. Employing the unique fluorescence properties of pyrene group, we designed and synthesized a sensitive fluorescence-based probe not only to target 5-formylcytosine (5fC) sites, but also to distinguish symmetric from asymmetric 5fC sites in the double stranded DNA context during TET-dependent 5mC oxidation process. Using this novel probe, we revealed dominant levels of symmetric 5fC among total 5fC sites during in vitro TET-dependent 5mC oxidation and novel mechanistic insights into the TET-dependent 5mC oxidation in the mCpG context.

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