Distinct catalytic and non-catalytic roles of ARGONAUTE4 in RNA-directed DNA methylation

Yijun Qi, Xingyue He, Xiu-Jie Wang, Oleksiy Kohany, Jerzy Jurka and Gregory J. Hannon ()
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Yijun Qi: Watson School of Biological Sciences and Howard Hughes Medical Institute
Xingyue He: Watson School of Biological Sciences and Howard Hughes Medical Institute
Xiu-Jie Wang: Institute of Genetics and Developmental Biology, Chinese Academy of Sciences
Oleksiy Kohany: Genetic Information Research Institute
Jerzy Jurka: Genetic Information Research Institute
Gregory J. Hannon: Watson School of Biological Sciences and Howard Hughes Medical Institute

Nature, 2006, vol. 443, issue 7114, 1008-1012

Abstract: Abstract DNA methylation has important functions in stable, transcriptional gene silencing, immobilization of transposable elements and genome organization1. In Arabidopsis, DNA methylation can be induced by double-stranded RNA through the RNA interference (RNAi) pathway, a response known as RNA-directed DNA methylation2. This requires a specialized set of RNAi components, including ARGONAUTE4 (AGO4)3,4,5,6. Here we show that AGO4 binds to small RNAs including small interfering RNAs (siRNAs) originating from transposable and repetitive elements, and cleaves target RNA transcripts. Single mutations in the Asp-Asp-His catalytic motif of AGO4 do not affect siRNA-binding activity but abolish its catalytic potential. siRNA accumulation and non-CpG DNA methylation at some loci require the catalytic activity of AGO4, whereas others are less dependent on this activity. Our results are consistent with a model in which AGO4 can function at target loci through two distinct and separable mechanisms. First, AGO4 can recruit components that signal DNA methylation in a manner independent of its catalytic activity. Second, AGO4 catalytic activity can be crucial for the generation of secondary siRNAs that reinforce its repressive effects.

Date: 2006
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DOI: 10.1038/nature05198

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