ROS3 is an RNA-binding protein required for DNA demethylation in Arabidopsis.
ROS3 is an RNA-binding protein required for DNA demethylation in Arabidopsis.
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DOI：
10.1038/nature07305
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发表时间：
2008-10-30
期刊：
影响因子：
64.8
通讯作者：
Zhu, Jian-Kang
中科院分区：
文献类型：
--
作者：
Zheng, Xianwu;Pontes, Olga;Zhu, Jianhua;Miki, Daisuke;Zhang, Fei;Li, Wen-Xue;Iida, Kei;Kapoor, Avnish;Pikaard, Craig S.;Zhu, Jian-Kang
DNA甲基化是多种生物中转录基因沉默（TGS）的重要表观遗传标记。最近的研究表明，许多基因的甲基化状态是由甲基化和去甲基化动态调节的。在拟南芥中，活性DNA去甲基化是由5-甲基胞嘧啶DNA糖基化酶的ROS 1亚家族通过碱基切除修复途径介导的。这些去甲基化酶在消除DNA甲基化和防止靶基因的TGS中起关键作用。然而，尚不清楚脱甲基酶如何靶向特定序列。我们在这里报告的识别ROS 3，一个重要的调节DNA去甲基化，包含一个RNA识别基序。ros 3突变体和ros 1 ros 3双突变体的分析表明，ROS 3与ROS 1在相同的遗传途径中起作用，以防止DNA超甲基化和TGS。凝胶迁移率变动分析和植物提取物中ROS 3免疫沉淀物的分析表明，ROS 3在体外和体内与小RNA结合。免疫染色显示，ROS 3和ROS 1蛋白共定位在分散在整个细胞核的离散病灶。这些结果证明了ROS 3在防止DNA超甲基化中的关键作用，并表明ROS 1的DNA去甲基化可能由与ROS 3结合的RNA指导。
DNA methylation is an important epigenetic mark for transcriptional gene silencing (TGS) in diverse organisms. Recent studies suggest that the methylation status of a number of genes is dynamically regulated by methylation and demethylation. In Arabidopsis, active DNA demethylation is mediated by the ROS1 (repressor of silencing 1) subfamily of 5-methylcytosine DNA glycosylases through a base excision repair pathway. These demethylases play critical roles in erasing DNA methylation and preventing TGS of target genes. However, it is not known how the demethylases are targeted to specific sequences. We report here the identification of ROS3, an essential regulator of DNA demethylation that contains an RNA recognition motif. Analysis of ros3 mutant and ros1ros3 double mutant suggests that ROS3 acts in the same genetic pathway as ROS1 to prevent DNA hypermethylation and TGS. Gel mobility shift assays and analysis of ROS3 immunoprecipitate from plant extracts showed that ROS3 binds to small RNAs in vitro and in vivo. Immunostaining shows that ROS3 and ROS1 proteins colocalize in discrete foci dispersed throughout the nucleus. These results demonstrate a critical role for ROS3 in preventing DNA hypermethylation and suggest that DNA demethylation by ROS1 may be guided by RNAs bound to ROS3.
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