Sequence-specific hypomethylation of the tobacco genome induced with dihydroxypropyladenine, ethionine and 5-azacytidine

Higher plant DNA is methylated at CG and CNG targets. In this study we have investigated the tobacco methylation system in tissue culture using the methylation inhibitors 5-azacytidine (5-azaC), dihydroxypropyladenine (DHPA) and ethionine (Ethi), and methylation-sensitive restriction endonucleases H...

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Bibliographic Details
Published in:FEBS letters 1994-10, Vol.353 (3), p.309-311
Main Authors: Kovarik, A, Koukalova, B, Holy, A, Bezdek, M
Format: Article
Language:English
Subjects:
Adenine - analogs & derivatives
Adenine - pharmacology
Adenosylhomocysteinase
Azacitidine - pharmacology
Cells, Cultured
CG and CNG DNA sequence
Deoxyribonucleases, Type II Site-Specific
DNA methylation inhibitor
DNA, Plant - metabolism
DNA-Cytosine Methylases - metabolism
Ethionine - pharmacology
Genome, Plant
Hydrolases - antagonists & inhibitors
Methylation - drug effects
Nicotiana - genetics
Nicotiana tabacum
Plants, Toxic
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Summary:Higher plant DNA is methylated at CG and CNG targets. In this study we have investigated the tobacco methylation system in tissue culture using the methylation inhibitors 5-azacytidine (5-azaC), dihydroxypropyladenine (DHPA) and ethionine (Ethi), and methylation-sensitive restriction endonucleases HpaII, MspI, HhaI, EcoRII, ScrFI, and Fnu4HI. Surprisingly, CAG/CTG sequences, contrary to CG doublets and CCG/CGG triplets, appeared to be refractory to the inhibitory effect of 5-azaC. Thus 5-azaC cannot be considered a general inhibitor of DNA methylation in tobacco cells. On the other hand, DHPA, the inhibitor of S-adenosylhomocysteine (SAH) hydrolase, and Ethi caused hypomethylation of both CAG/CTG and CCG/CGG triplets but not of the CG doublets. The sensitivity of triplet-specific methylation to the inhibition of SAH hydrolase suggests the possibility that plant-specific DNA methylation at CNG targets might be modulated by alterations of the SAH/S-Adenosyhnethionine ratio in plant cells.
ISSN:0014-5793
1873-3468
DOI:10.1016/0014-5793(94)01048-X