老鼠Tet1蛋白能氧化单链脱氧核糖核酸上的5-甲基胞嘧啶到5-羟甲基胞嘧啶和5-羧基胞嘧啶

doi:10.6023/A12090619

摘要/Abstract

5-甲基胞嘧啶在哺乳动物细胞中具有广泛的作用. 而它被双脱氧家族Tet蛋白氧化所得的产物5-羟甲基胞嘧啶、5-醛基胞嘧啶和5-羧基胞嘧啶也被证明在细胞发育和5-甲基胞嘧啶动态平衡调控中具有关键的作用. 已有的研究结果表明, Tet蛋白能够识别双链DNA上的5-甲基胞嘧啶, 并将其氧化成5-羟甲基胞嘧啶、5-醛基胞嘧啶和5-羧基胞嘧啶. 我们通过质谱仪检测发现, 老鼠Tet1蛋白的DNA结合结构域还能识别和氧化单链DNA上的5-甲基胞嘧啶. 这一发现暗示我们, Tet蛋白家族不但具有已经发现的氧化双链DNA上5-甲基胞嘧啶的功能, 还有可能作用于DNA的复制及转录, 甚至具有氧化单链RNA上对应的甲基修饰碱基的能力.

关键词: Tet蛋白, 5-甲基胞嘧啶, 5-羟甲基胞嘧啶, 5-醛基胞嘧啶, 5-羧基胞嘧啶, 双链DNA, 单链DNA, 氧化

5-Methylcytosine (5mC) is known to play broad roles in mammalian biology. It can be enzymatically oxidized to 5-hydroxymethylcytosine (5hmC), 5-formylcytosine (5fC), and 5-carboxylcytosine (5caC) by a family of iron(II)/αKG-dependent dioxygenases Tet proteins. This process has been shown to play critical roles in mammalian cell development and dynamic methylation regulation. Previous research has shown that Tet proteins can oxidize 5mC on double-stranded DNA (dsDNA). Here, we expressed and purified the active domain of mouse Tet1 DNA from insect cell. By employing mass spectrometry, we show here that mtet1 (1367—2039) can also oxidize 5mC on single-stranded DNA (ssDNA), thus indicating its potential involvement in 5mC oxidation during replication or/and transcription, as well as the oxidation of corresponding modifications on single-stranded RNA in mammalian cells.

Key words: Tet, 5-methylcytosine (5mC), 5-hydroxymethylcytosine (5hmC), 5-formylcytosine (5fC), 5-carboxylcytosine (5caC), double-stranded DNA (dsDNA), single-stranded DNA (ssDNA), oxidation

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张良, 于淼, 何川. 老鼠Tet1蛋白能氧化单链脱氧核糖核酸上的5-甲基胞嘧啶到5-羟甲基胞嘧啶和5-羧基胞嘧啶[J]. 化学学报, 2012, 70(20): 2123-2126.

Zhang Liang, Yu Miao, He Chuan. Mouse Tet1 Protein can Oxidize 5mC to 5hmC and 5caC on Single-stranded DNA[J]. Acta Chimica Sinica, 2012, 70(20): 2123-2126.

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