A protein complex required for polymerase V transcripts and RNA- directed DNA methylation in Arabidopsis.
DNA methylation is an epigenetic modification associated with gene silencing. In Arabidopsis, DNA methylation is established by DOMAINS REARRANGED METHYLTRANSFERASE 2 (DRM2), which is targeted by small interfering RNAs through a pathway termed RNA-directed DNA methylation (RdDM). Recently, RdDM was shown to require intergenic noncoding (IGN) transcripts that are dependent on the Pol V polymerase. These transcripts are proposed to function as scaffolds for the recruitment of downstream RdDM proteins, including DRM2, to loci that produce both siRNAs and IGN transcripts. However, the mechanism(s) through which Pol V is targeted to specific genomic loci remains largely unknown. Through affinity purification of two known RdDM components, DEFECTIVE IN RNA-DIRECTED DNA METHYLATION 1 (DRD1) and DEFECTIVE IN MERISTEM SILENCING 3 (DMS3), we found that they copurify with each other and with a novel protein, RNA-DIRECTED DNA METHYLATION 1 (RDM1), forming a complex we term DDR. We also found that DRD1 copurified with Pol V subunits and that RDM1, like DRD1 and DMS3, is required for the production of Pol V-dependent transcripts. These results suggest that the DDR complex acts in RdDM at a step upstream of the recruitment or activation of Pol V.
Law
Julie A
JA
Department of Molecular Cell and Developmental Biology, University of California at Los Angeles, Los Angeles, CA 90095, USA.
Ausin
Israel
I
Johnson
Lianna M
LM
Vashisht
Ajay A
AA
Zhu
Jian-Kang
JK
Wohlschlegel
James A
JA
Jacobsen
Steven E
SE
D017360:Arabidopsis / Q000235:genetics / Q000378:metabolism; D029681:Arabidopsis Proteins / Q000235:genetics / Q000378:metabolism*; D002868:Chromosomal Proteins, Non-Histone / Q000235:genetics / Q000378:metabolism; D019175:DNA Methylation; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D004259:DNA-Directed DNA Polymerase / Q000235:genetics / Q000378:metabolism*; D012321:DNA-Directed RNA Polymerases / Q000235:genetics / Q000378:metabolism; D018506:Gene Expression Regulation, Plant; D007527:Isoenzymes / Q000235:genetics / Q000378:metabolism; D013058:Mass Spectrometry; D030821:Plants, Genetically Modified; D018749:RNA, Plant / Q000235:genetics / Q000378:metabolism*
D029681:Arabidopsis Proteins; D002868:Chromosomal Proteins, Non-Histone; C529025:DMS3 protein, Arabidopsis; D004268:DNA-Binding Proteins; D007527:Isoenzymes; C549271:RDM1 protein, Arabidopsis; D018749:RNA, Plant; D012321:DNA-Directed RNA Polymerases; C504888:DRD1 protein, Arabidopsis; D004259:DNA-Directed DNA Polymerase
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