A DNA 3' phosphatase functions in active DNA demethylation in Arabidopsis.

A DNA 3' phosphatase functions in active DNA demethylation in Arabidopsis.
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DOI:
10.1016/j.molcel.2011.11.034
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发表时间:
2012-02-10
期刊:
影响因子:
16
通讯作者:
Zhu JK
Zhu JK
中科院分区:
生物学1区
文献类型:
--
作者:
Martínez-Macías MI;Qian W;Miki D;Pontes O;Liu Y;Tang K;Liu R;Morales-Ruiz T;Ariza RR;Roldán-Arjona T;Zhu JK

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DNA甲基化是甲基化和去甲基化反应共同作用下形成的重要表观遗传标记。植物使用碱基切除修复途径进行活性DNA去甲基化。去除5-甲基胞嘧啶后,拟南芥DNA糖基化酶/裂解酶ROS1切开DNA主干,部分产物有一个单核苷酸缺口,两侧是3 ' -和5 ' -磷酸端。在这里,我们展示了DNA磷酸酶ZDP去除阻断3 ' -磷酸,允许随后的DNA聚合和连接步骤完成修复反应。ZDP和ROS1在体外相互作用,并在体内核质病灶中共定位。zdp突变植株的提取物在体外不能完成DNA去甲基化,突变导致DNA超甲基化和一个报告基因的转录沉默。zdp突变株的全基因组甲基化分析鉴定出数百个高甲基化的内源位点。我们的研究结果表明,ZDP在ROS1活性DNA去甲基化途径的一个分支中发挥下游作用。
DNA methylation is an important epigenetic mark established by the combined actions of methylation and demethylation reactions. Plants use a base excision repair pathway for active DNA demethylation. After 5-methylcytosine removal, the Arabidopsis DNA glycosylase/lyase ROS1 incises the DNA backbone and part of the product has a single-nucleotide gap flanked by 3′- and 5′-phosphate termini. Here we show that the DNA phosphatase ZDP removes the blocking 3′-phosphate, allowing subsequent DNA polymerization and ligation steps needed to complete the repair reactions. ZDP and ROS1 interact in vitro and co-localize in vivo in nucleoplasmic foci. Extracts from zdp mutant plants are unable to complete DNA demethylation in vitro, and the mutations cause DNA hypermethylation and transcriptional silencing of a reporter gene. Genome-wide methylation analysis in zdp mutant plants identified hundreds of hypermethylated endogenous loci. Our results show that ZDP functions downstream of ROS1 in one branch of the active DNA demethylation pathway.
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