IDN1 and IDN2 are required for de novo DNA methylation in Arabidopsis thaliana - PubMed (original) (raw)

. 2009 Dec;16(12):1325-7.

doi: 10.1038/nsmb.1690. Epub 2009 Nov 15.

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IDN1 and IDN2 are required for de novo DNA methylation in Arabidopsis thaliana

Israel Ausin et al. Nat Struct Mol Biol. 2009 Dec.

Abstract

DNA methylation is an epigenetic mark affecting genes and transposons. Screening for mutants that fail to establish DNA methylation yielded two we termed "involved in de novo" (idn) 1 and 2. IDN1 encodes DMS3, an SMC-related protein, and IDN2 encodes a previously unknown double-stranded RNA-binding protein with homology to SGS3. IDN1 and IDN2 control de novo methylation and small interfering RNA (siRNA)-mediated maintenance methylation and are components of the RNA-directed DNA methylation pathway.

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Figures

Figure 1

Figure 1. De novo methylation phenotype of idn mutants

a) Flowering time measured as total number of leaves produced by wild type, idn mutants and FWA transformed T1 plants under long day conditions. Error bars depict standard deviation. b) Methylation status of wild type and idn mutants at endogenous and transgenic FWA. c) SDC expression levels in idn mutants before and after SDC transformation. Hybridization with a UBQ10 probe is shown as loading control. Panel below shows methylation status of wild type and idn mutants at transgenic SDC.

Figure 2

Figure 2. Maintenance methylation phenotype of idn mutants

a) Methylation sensitive enzyme Southern hybridization assay at 5S and MEA-ISR loci, and HaeIII cutting assay at AtSN1. HaeIII, is blocked by C methylation in GGCC context, HpaII is blocked by C methylation in CCGG context and MspI is blocked by methylation of the external C in CCGG context. b) Methylation status of wild type, idn and idn cmt3–11 double mutants at endogenous SDC and methylation status of wild type and idn mutants at MEA-ISR. c) Morphological and molecular phenotype of idn cmt3–11 double mutants.

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