Identification of Sequence Specificity of 5‐Methylcytosine Oxidation by Tet1 Protein with High‐Throughput Sequencing
Kizaki S., Chandran A., Sugiyama H.
AbstractTet (ten‐eleven translocation) family proteins have the ability to oxidize 5‐methylcytosine (mC) to 5‐hydroxymethylcytosine (hmC), 5‐formylcytosine (fC), and 5‐carboxycytosine (caC). However, the oxidation reaction of Tet is not understood completely. Evaluation of genomic‐level epigenetic changes by Tet protein requires unbiased identification of the highly selective oxidation sites. In this study, we used high‐throughput sequencing to investigate the sequence specificity of mC oxidation by Tet1. A 6.6×104‐member mC‐containing random DNA‐sequence library was constructed. The library was subjected to Tet‐reactive pulldown followed by high‐throughput sequencing. Analysis of the obtained sequence data identified the Tet1‐reactive sequences. We identified mCpG as a highly reactive sequence of Tet1 protein.