Functional characterization of the RNA guided DRM2 DNA methyltransferase. Research done in the Faculty of Biology UW and IBB PAS. M.Sc. Thesis Supervisor: prof. dr hab. Ewa Bartnik.

Abstract

DNA methylation in plants is in part controlled by small RNAs. A key player in RNA directed DNA methylation (RdDM) is Domain Rearranged Methyltransferase 2 (DRM2). In addition to its catalytic domain, which resembles the catalytic domain of animal DNA methyltransferases, the protein has several UBA domains of unknown biochemical significance. Genetic experiments on Arabidopsis thaliana indicate that a DRM2 variant lacking all UBA domains is similarly incapacitated as a catalytic mutant. The aim of this work was to elucidate the role of the UBA domains in DRM2. Towards this end, expression plasmids were generated, which should direct expression of DRM2 variants lacking individual UBA domains only after Agrobacterium mediated transformation. Unfortunately, protein expression of the deletion constructs was not observed, even though a reporter control protein was expressed in parallel experiments.