Expression of ZmMET1, a gene encoding a DNA methyltransferase from maize, is associated not only with DNA replication in actively proliferating cells, but also with altered DNA methylation status in cold-stressed quiescent cells.
A cDNA fragment encoding part of a DNA methyltransferase was isolated from maize. The putative amino acid sequence identically matched that deduced from a genomic sequence in the database (accession no. AF063403), and the corresponding gene was designated as ZmMET1. Bacterially expressed ZmMET1 actively methylated DNA in vitro. Transcripts of ZmMET1 could be shown to exclusively accumulate in actively proliferating cells of the meristems of mesocotyls and root apices, suggesting ZmMET1 expression to be associated with DNA replication. This was confirmed by simultaneous decrease of transcripts of ZmMET1 and histone H3, a marker for DNA replication, in seedlings exposed to wounding, desiccation and salinity, all of which suppress cell division. Cold stress also depressed both transcripts in root tissues. In contrast, however, accumulation of ZmMET1 transcripts in shoot mesocotyls was not affected by cold stress, whereas those for H3 sharply decreased. Such a differential accumulation of ZmMET1 transcripts was consistent with ZmMET1 protein levels as revealed by western blotting. Expression of ZmMET1 is thus coexistent, but not completely dependent on DNA replication. Southern hybridization analysis with a methylation-sensitive restriction enzyme revealed that cold treatment induced demethylation of DNA in the Ac/Ds transposon region, but not in other genes, and that such demethylation primarily occurred in roots. These results suggested that the methylation level was decreased selectively by cold treatment, and that ZmMET1 may, at least partly, prevent such demethylation.
Steward
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Research and Education Center for Genetic Information, Nara Institute of Science and Technology, 8916-5 Takayama-cho Ikoma, Nara 630-0101 Japan.
Kusano
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T
Sano
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H
D015139:Blotting, Southern; D015153:Blotting, Western; D002455:Cell Division; D003001:Cloning, Molecular; D003080:Cold Temperature; D004248:DNA (Cytosine-5-)-Methyltransferases / Q000378:metabolism*; D004249:DNA Damage / Q000187:drug effects / Q000235:genetics; D019175:DNA Methylation; D015254:DNA Modification Methylases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D004261:DNA Replication / Q000187:drug effects; D005020:Ethyl Methanesulfonate / Q000494:pharmacology; D018506:Gene Expression Regulation, Plant / Q000187:drug effects; D017403:In Situ Hybridization; D008969:Molecular Sequence Data; D009928:Organ Specificity; D010935:Plant Diseases / Q000235:genetics; D018514:Plant Structures / Q000166:cytology / Q000187:drug effects / Q000201:enzymology / Q000235:genetics; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics; D018749:RNA, Plant / Q000032:analysis / Q000235:genetics; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D003313:Zea mays / Q000166:cytology* / Q000187:drug effects / Q000201:enzymology / Q000235:genetics*
D012333:RNA, Messenger; D018749:RNA, Plant; D011994:Recombinant Proteins; D005020:Ethyl Methanesulfonate; D015254:DNA Modification Methylases; D004248:DNA (Cytosine-5-)-Methyltransferases