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The Temperature-Dependent Change in Methylation of the Antirrhinum Transposon Tam3 Is Controlled by the Activity of Its Transposase^[W]

^a Laboratory of Genetic Engineering, Graduate School of Agriculture, Hokkaido University, Sapporo 060-8589, Japan
^b Laboratory of Plant Breeding, Graduate School of Agriculture, Hokkaido University, Sapporo 060-8589, Japan
^c Department of Cell and Developmental Biology, John Innes Centre, Norwich Research Park, Norwich, Norfolk NR4 7UH, United Kingdom

² To whom correspondence should be addressed. E-mail kishima{at}abs.agr.hokudai.ac.jp; fax 81-11-706-4934.

The Antirrhinum majus transposon Tam3 undergoes low temperature–dependent transposition (LTDT). Growth at 15°C permits transposition, whereas growth at 25°C strongly suppresses it. The degree of Tam3 DNA methylation is altered somatically and positively correlated with growth temperature, an exceptional epigenetic system in plants. Using a Tam3-inactive line, we show that methylation change depends on Tam3 activity. Random binding site selection analysis and electrophoretic mobility shift assays revealed that the Tam3 transposase (TPase) binds to the major repeat in the subterminal regions of Tam3, the site showing the biggest temperature-dependent change in methylation state. Methylcytosines in the motif impair the binding ability of the TPase. Proteins in a nuclear extract from plants grown at 15°C but not 25°C bind to this motif in Tam3. The decrease in Tam3 DNA methylation at low temperature also requires cell division. Thus, TPase binding to Tam3 occurs only during growth at low temperature and immediately after DNA replication, resulting in a Tam3-specific decrease in methylation of transposon DNA. Consequently, the Tam3 methylation level in LTDT is regulated by Tam3 activity, which is dependent on the ability of its TPase to bind DNA and affected by growth temperature. Thus, the methylation/demethylation of Tam3 is the consequence, not the cause, of LTDT.

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