RNA-directed DNA methylation in Arabidopsis

We describe this system and the impact of several mutations that impair DNA methylation andor possible chromatin remodeling processes

W. Aufsatz; M. F. Mette; J. van der Winden; A. J. M. Matzke; M. Matzke

2002

Scholarcy highlights

  • In plants, double-stranded RNA that is processed to short RNAs Ϸ21–24 nt in length can trigger two types of epigenetic gene silencing
  • A promoter double-stranded RNA-mediated trans-silencing system based on the nopaline synthase promoter was originally established in tobacco and has served as a model for setting up a similar system in Arabidopsis
  • The target line was retransformed with a silencing construct, which contains a NOSpro inverted repeats under the control of the 35S promoter of cauliflower mosaic virus together with a gene encoding resistance to hygromycin B driven by the 19S promoter of cauliflower mosaic virus
  • The silencing locus produces NOSpro dsRNA that is processed into short RNAs Ϸ21–24 nucleotides in length, similar to those observed in tobacco transformed with the same construct
  • A NOSpro dsRNA transcribed from a NOSpro IR at the silencing locus is processed to short RNAs 21–24 nt in length
  • We report here an analysis of methylation of the target NOSpro and the NOSpro IR at the silencing locus in the presence and absence of NOSpro dsRNA, and in several mutants deficient in DNA methylation andor putative chromatin remodeling proteins
  • We thank Eric Richards for the met1 mutant and for probes to analyze methylation in centromeric and ribosomal DNA repeats; Ortrun Mittelsten Scheid for the som8 allele of ddm1; Jurek Paszkowski for mom1; Ann Depicker for supplying a Cre-expressing T-DNA construct; and Steve Jacobsen and Xiaofeng Cao for a collaboration on cmt3 and drm2 mutants

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