Identification of Sequence Specificity of 5-Methylcytosine Oxidation by Tet1 Protein with High-Throughput Sequencing
Identification of Sequence Specificity of 5-Methylcytosine Oxidation by Tet1 Protein with High-Throughput Sequencing
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DOI：
10.1002/cbic.201500646
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发表时间：
2016-03-01
期刊：
影响因子：
3.2
通讯作者：
Sugiyama, Hiroshi
中科院分区：
文献类型：
--
作者：
Kizaki, Seiichiro;Chandran, Anandhakumar;Sugiyama, Hiroshi
泰特（ten-eleven易位）家族蛋白具有将5-甲基胞嘧啶（mC）氧化为5-羟甲基胞嘧啶（hmC）、5-甲酰基胞嘧啶（fC）和5-羧基胞嘧啶（caC）的能力。然而，泰特的氧化反应还没有完全了解。通过泰特蛋白评估基因组水平的表观遗传变化需要无偏地鉴定高选择性氧化位点。在这项研究中，我们使用高通量测序来研究Tet 1氧化mC的序列特异性。构建了6.6 × 10（4）个成员的含mC的随机DNA序列文库。对文库进行Tet-reactive pulldown，然后进行高通量测序。对获得的序列数据进行分析，鉴定了Tet 1反应性序列。我们将mCpG鉴定为Tet 1蛋白的高反应性序列。
Tet (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 6.6x10(4)-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.