title: Effective, homogeneous and transient interference with cytosine methylation in plant genomic DNA by zebularine.
abstract: Covalent modification by methylation of cytosine residues represents an important epigenetic hallmark. While sequence analysis after bisulphite conversion allows correlative analyses with single-base resolution, functional analysis by interference with DNA methylation is less precise, due to the complexity of methylation enzymes and their targets. A cytidine analogue, 5-azacytidine, is frequently used as an inhibitor of DNA methyltransferases, but its rapid degradation in aqueous solution is problematic for culture periods of longer than a few hours. Application of zebularine, a more stable cytidine analogue with a similar mode of action that is successfully used as a methylation inhibitor in Neurospora and mammalian tumour cell lines, can significantly reduce DNA methylation in plants in a dose-dependent and transient manner independent of sequence context. Demethylation is connected with transcriptional reactivation and partial decondensation of heterochromatin. Zebularine represents a promising new and versatile tool for investigating the role of DNA methylation in plants with regard to transcriptional control, maintenance and formation of (hetero-) chromatin.
authors.affiliation: Gregor Mendel Institute of Molecular Plant Biology, Austrian Academy of Sciences, Dr Bohr-Gasse 3, 1030 Vienna, Austria.
authors.forename: Tuncay
authors.initials: T
authors.lastname: Baubec
authors.forename: Ales
authors.initials: A
authors.lastname: Pecinka
authors.forename: Wilfried
authors.initials: W
authors.lastname: Rozhon
authors.forename: Ortrun
authors.initials: O
authors.lastname: Mittelsten Scheid
mesh_terms: D017360:Arabidopsis / Q000235:genetics / Q000254:growth & development / Q000378:metabolism; D003562:Cytidine / Q000031:analogs & derivatives* / Q000494:pharmacology; D003596:Cytosine / Q000378:metabolism*; D019175:DNA Methylation / Q000187:drug effects*; D018744:DNA, Plant / Q000187:drug effects*; D003841:Deoxycytidine / Q000031:analogs & derivatives / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D018745:Genome, Plant; D000455:Medicago sativa / Q000235:genetics / Q000254:growth & development / Q000378:metabolism
chemical_list: D018744:DNA, Plant; D003841:Deoxycytidine; D003562:Cytidine; C009131:pyrimidin-2-one beta-ribofuranoside; D003596:Cytosine; C016569:5-methyldeoxycytidine