The role of MET1 in RNA-directed de novoand maintenance methylation of CG dinucleotides

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Abstract
A genetic screen for mutants defective in RNA-directed DNA methylation and transcriptional silencing of the constitutive nopaline synthase (NOS) promoter in Arabidopsisidentified two independent mutations in the gene encoding the DNA methyltransferase MET1. Both mutant alleles are disrupted structurally in the MET1 catalytic domain, suggesting that they are complete loss of function alleles. Experiments designed to test the effect of a met1mutation on both RNA-directed de novoand maintenance methylation of the target NOS promoter revealed in each case approximately wild type levels of non-CG methylation together with significant reductions of CG methylation. These results confirm a requirement for MET1 to maintain CG methylation induced by RNA. In addition, the failure to establish full CG methylation in met1mutants, despite normal RNA-directed de novo methylation of Cs in other sequence contexts, indicates that MET1 is required for full de novomethylation of CG dinucleotides. We discuss MET1 as a site-specific DNA methyltransferase that is able to maintain CG methylation during DNA replication and contribute to CG de novomethylation in response to RNA signals.