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Genetics, Vol. 157, 1723-1733, April 2001, Copyright © 2001

Regulation of Activator/Dissociation Transposition by Replication and DNA Methylation

Francesca Rosa and Reinhard Kunzea
a Institut für Genetik und Mikrobiologie, Universität München, 80638 Munich, Germany

Corresponding author: Reinhard Kunze, Botanisches Institut II, Universität zu Köln, Gyrhofstrasse 15, 50931 Cologne, Germany., reinhard.kunze{at}uni-koeln.de (E-mail)

Communicating editor: V. SUNDARESAN

In maize the transposable elements Activator/Dissociation (Ac/Ds) transpose shortly after replication from one of the two resulting chromatids ("chromatid selectivity"). A model has been suggested that explains this phenomenon as a consequence of different affinity for Ac transposase binding to holo-, hemi-, and unmethylated transposon ends. Here we demonstrate that in petunia cells a holomethylated Ds is unable to excise from a nonreplicating vector and that replication restores excision. A Ds element hemi-methylated on one DNA strand transposes in the absence of replication, whereas hemi-methylation of the complementary strand causes a >6.3-fold inhibition of Ds excision. Consistently in the active hemi-methylated state, the Ds ends have a high binding affinity for the transposase, whereas binding to inactive ends is strongly reduced. These results provide strong evidence for the above-mentioned model. Moreover, in the absence of DNA methylation, replication enhances Ds transposition in petunia protoplasts >8-fold and promotes formation of a predominant excision footprint. Accordingly, replication also has a methylation-independent regulatory effect on transposition.




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