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Plant and Cell Physiology Advance Access published online on April 5, 2007
Plant and Cell Physiology, doi:10.1093/pcp/pcm043
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© The Author 2007. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For Permissions, please e-mail: journals.permissions@oxfordjournals.org

Arabidopsis thaliana 5S ribosomal RNA genes regulation.

Isabelle Vaillant1, Sylvie Tutois, Claudine Cuvillier, Ingo Schubert2 and Sylvette Tourmente*

1 Present address: University of Geneva, Laboratory of Plant Genetics Sciences III, 30 Quai Ernest-Ansermet, CH-1211 Genève 4, Switzerland.
2 Leibniz- Institute of Plant Genetics and Crop Plant Research (IPK), Corrensstrasse 3, D-06466 Gatersleben, Germany (I.S.).

*Corresponding author: Sylvette Tourmente. Unité Mixte de Recherche CNRS 6547 BIOMOVE, Université Blaise Pascal, 24 Avenue des Landais, 63177 Aubière Cedex, France. Tel: (33) 4 73 40 74 01; FAX: (33) 4 73 40 77 77; Email: sylvette.tourmente{at}univ-bpclermont.fr


  Abstract

The Arabidopsis thaliana genome comprises around 1000 copies of 5S rRNA genes encoding both major and minor 5S rRNAs. In mature wild type leaves, the minor 5S rRNA genes are silent. Using different mutants of DNA methyltransferases (met1, cmt3 and met1 cmt3), components of the RNAi pathway (ago4) or post-translational histone modifier (hda6/sil1), we show that the corresponding proteins are needed to maintain proper methylation patterns at heterochromatic 5S rDNA repeats. Using RT-PCR and cytological analyses, we report that decrease of 5S rDNA methylation at CG or CNG sites in these mutants lead to the release of 5S rRNA genes silencing which occured without detectable changes of the 5S rDNA chromatin structure. Inspite of severely reduced DNA methylation, the met1 cmt3 double mutant revealed no increase in minor 5S rRNA transcripts.

Furthermore, the release of minor 5S rDNA silencing can be achieved without an increased formation of euchromatic loops by 5S rDNA, and is independent from the global heterochromatin content.

Additionally, FISH with centromeric 180-bp repeats confirmed that these highly repetitive sequences inspite of their elevated transcriptional activity in the DNA methyltransferase mutants (met1, cmt3 and met1 cmt3) remain within chromocenters of the mutant nuclei.

Keywords: Arabidopsis thaliana - 5S rDNA - DNA methylation - Transcription


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