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An Effector of RNA-Directed DNA Methylation in Arabidopsis Is an ARGONAUTE 4-and RNA-Binding Protein

机译:拟南芥中RNA定向DNA甲基化的效应是ARGONAUTE 4和RNA结合蛋白。

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摘要

DNA methylation is a conserved epigenetic mark in plants and mammals. In Arabidopsis, DNA methylation can be triggered by small interfering RNAs (siRNAs) through an RNA-directed DNA methylation (RdDM) pathway. Here, we report the identification of an RdDM effector, KTF1. Loss-of-function mutations in KTF1 reduce DNA methylation and release the silencing of RdDM target loci without abolishing the siRNA triggers. KTF1 has similarity to the transcription elongation factor SPT5 and contains a C-terminal extension rich in GW/WG repeats. KTF1 colocalizes with ARGONAUTE 4 (AGO4) in punctate nuclear foci and binds AGO4 and RNA transcripts. Our results suggest KTF1 as an adaptor protein that binds scaffold transcripts generated by Pol V and recruits AGO4 and AGO4-bound siRNAs to form an RdDM effector complex. The dual interaction of an effector protein with AGO and small RNA target transcripts may be a general feature of RNA-silencing effector complexes.
机译:DNA甲基化是植物和哺乳动物中保守的表观遗传标记。在拟南芥中,小分子干扰RNA(siRNA)可通过RNA定向DNA甲基化(RdDM)途径触发DNA甲基化。在这里,我们报告了RdDM效应子KTF1的鉴定。 KTF1中的功能丧失突变可降低DNA甲基化并释放RdDM目标基因座的沉默,而不会消除siRNA触发因素。 KTF1与转录延伸因子SPT5具有相似性,并且包含一个富含GW / WG重复序列的C端延伸。 KTF1在点状核灶中与ARGONAUTE 4(AGO4)共定位,并结合AGO4和RNA转录本。我们的结果表明,KTF1是一种衔接蛋白,可结合Pol V产生的支架转录物并募集AGO4和AGO4结合的siRNA,形成RdDM效应子复合物。效应蛋白与AGO和小RNA靶转录本的双重相互作用可能是RNA沉默效应复合物的普遍特征。

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