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ROS3 is an RNA-binding protein required for DNA demethylation in Arabidopsis

机译：ROS3是拟南芥DNA脱甲基化所需的RNA结合蛋白

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摘要

DNA methylation is an important epigenetic mark for transcrip-tional 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 (repres-sor of silencing 1) subfamily of 5-methylcytosine DNA glycosylases through a base excision repair pathway. These demethylases have 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. Here we report the identification of ROS3, an essential regulator of DNA demethylation that contains an RNA recognition motif. Analysis of ros3 mutants and rosl ros3 double mutants 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 shows that ROS3 binds to small RNAs in vitro and in vivo. Immunostaining shows that ROS3 and ROS1 proteins co-localize 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.
机译：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由甲基化和脱甲基化动态调节。在拟南芥中，活性DNA去甲基化是通过碱基切除修复途径由5-甲基胞嘧啶DNA糖基化酶的ROS1（沉默1的代表）亚家族介导的。这些脱甲基酶在消除DNA甲基化和防止靶基因的TGS中起关键作用。然而，不知道脱甲基酶如何靶向特定序列。在这里，我们报告鉴定的ROS3，DNA脱甲基的重要调节器，其中包含一个RNA识别基序。对ros3突变体和rossl ros3双重突变体的分析表明，ROS3与ROS1在相同的遗传途径中起作用，以防止DNA过度甲基化和TGS。凝胶迁移率迁移分析和植物提取物中ROS3免疫沉淀物的分析表明，ROS3在体外和体内与小RNA结合。免疫染色显示，ROS3和ROS1蛋白共定位在分散于整个细胞核的离散病灶中。这些结果证明了ROS3在防止DNA过度甲基化中的关键作用，并表明ROS1的DNA去甲基化可能受结合到ROS3的RNA的引导。

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