Effective, homogeneous and transient interference with cytosine methylation in plant genomic DNA by zebularine.
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.
Baubec
Tuncay
T
Gregor Mendel Institute of Molecular Plant Biology, Austrian Academy of Sciences, Dr Bohr-Gasse 3, 1030 Vienna, Austria.
Pecinka
Ales
A
Rozhon
Wilfried
W
Mittelsten Scheid
Ortrun
O
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
D018744:DNA, Plant; D003841:Deoxycytidine; D003562:Cytidine; C009131:pyrimidin-2-one beta-ribofuranoside; D003596:Cytosine; C016569:5-methyldeoxycytidine