Evidence for ARGONAUTE4-DNA interactions in RNA-directed DNA methylation in plants

Genes Dev. 2016 Dec 1;30(23):2565-2570. doi: 10.1101/gad.289553.116. Epub 2016 Dec 16.


RNA polymerase V (Pol V) long noncoding RNAs (lncRNAs) have been proposed to guide ARGONAUTE4 (AGO4) to chromatin in RNA-directed DNA methylation (RdDM) in plants. Here, we provide evidence, based on laser UV-assisted zero-length cross-linking, for functionally relevant AGO4-DNA interaction at RdDM targets. We further demonstrate that Pol V lncRNAs or the act of their transcription are required to lock Pol V holoenzyme into a stable DNA-bound state that allows AGO4 recruitment via redundant glycine-tryptophan/tryptophan-glycine AGO hook motifs present on both Pol V and its associated factor, SPT5L. We propose a model in which AGO4-DNA interaction could be responsible for the unique specificities of RdDM.

Keywords: Argonaute; DNA methylation; RdDM.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, N.I.H., Extramural

MeSH terms

  • Amino Acid Motifs / genetics
  • Arabidopsis Proteins / chemistry
  • Arabidopsis Proteins / genetics*
  • Arabidopsis Proteins / metabolism*
  • Argonaute Proteins / chemistry
  • Argonaute Proteins / genetics*
  • Argonaute Proteins / metabolism*
  • Chromatin / metabolism
  • DNA Methylation* / genetics
  • DNA, Plant / metabolism*
  • DNA-Directed RNA Polymerases / chemistry
  • DNA-Directed RNA Polymerases / genetics
  • DNA-Directed RNA Polymerases / metabolism
  • Gene Silencing
  • Models, Biological
  • RNA Interference
  • RNA, Plant / metabolism*
  • Transcription, Genetic / genetics


  • AGO4 protein, Arabidopsis
  • Arabidopsis Proteins
  • Argonaute Proteins
  • Chromatin
  • DNA, Plant
  • RNA, Plant
  • RNA polymerase V, Arabidopsis
  • DNA-Directed RNA Polymerases