title: Role of the DRM and CMT3 methyltransferases in RNA-directed DNA methylation.
abstract: RNA interference is a conserved process in which double-stranded RNA is processed into 21-25 nucleotide siRNAs that trigger posttranscriptional gene silencing. In addition, plants display a phenomenon termed RNA-directed DNA methylation (RdDM) in which DNA with sequence identity to silenced RNA is de novo methylated at its cytosine residues. This methylation is not only at canonical CpG sites but also at cytosines in CpNpG and asymmetric sequence contexts. In this report, we study the role of the DRM and CMT3 DNA methyltransferase genes in the initiation and maintenance of RdDM. Neither drm nor cmt3 mutants affected the maintenance of preestablished RNA-directed CpG methylation. However, drm mutants showed a nearly complete loss of asymmetric methylation and a partial loss of CpNpG methylation. The remaining asymmetric and CpNpG methylation was dependent on the activity of CMT3, showing that DRM and CMT3 act redundantly to maintain non-CpG methylation. These DNA methyltransferases appear to act downstream of siRNAs, since drm1 drm2 cmt3 triple mutants show a lack of non-CpG methylation but elevated levels of siRNAs. Finally, we demonstrate that DRM activity is required for the initial establishment of RdDM in all sequence contexts including CpG, CpNpG, and asymmetric sites.
authors.affiliation: Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, 917 Datun Road, Beijing 100101, China.
authors.forename: Xiaofeng
authors.initials: X
authors.lastname: Cao
authors.forename: Werner
authors.initials: W
authors.lastname: Aufsatz
authors.forename: Daniel
authors.initials: D
authors.lastname: Zilberman
authors.forename: M Florian
authors.initials: MF
authors.lastname: Mette
authors.forename: Michael S
authors.initials: MS
authors.lastname: Huang
authors.forename: Marjori
authors.initials: M
authors.lastname: Matzke
authors.forename: Steven E
authors.initials: SE
authors.lastname: Jacobsen
mesh_terms: D017360:Arabidopsis; D029681:Arabidopsis Proteins; D015152:Blotting, Northern; D015139:Blotting, Southern; D018899:CpG Islands / Q000235:genetics*; D019175:DNA Methylation; D015257:DNA-Cytosine Methylases / Q000235:genetics / Q000378:metabolism*; D020868:Gene Silencing; D008780:Methyltransferases / Q000235:genetics / Q000378:metabolism*; D034741:RNA, Small Interfering / Q000378:metabolism
chemical_list: D029681:Arabidopsis Proteins; D034741:RNA, Small Interfering; D015257:DNA-Cytosine Methylases; D008780:Methyltransferases; C429766:CMT3 protein, Arabidopsis; C470731:DRM1 protein, Arabidopsis; C470732:DRM2 protein, Arabidopsis