Plant Physiol. Drug Metab Dispos
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Published on May 6, 2009; 10.1104/pp.109.137968

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Received March 3, 2009
Accepted April 29, 2009

Processing of a dicistronic tRNA-snoRNA precursor: combined analysis in vitro and in vivo reveals alternate pathways and coupling to assembly of snoRNP

Nicolas Barbezier , Giusy Canino , Julie Rodor , Edouard Jobet , Julio Saez-Vasquez , Anita Marchfelder , and Manuel Echeverria *

Laboratoire Genome et Developpement des Plantes, UMR 5096 Universite de Perpignan-CNRS-IRD, 52, Av Paul Alduy, 66 860 Perpignan Cedex, France; Molekulare Botanik, Universitat Ulm, Albert-Allee 11, 89069 Ulm, Germany

* Corresponding author; email: manuel.echeverria{at}univ-perp.fr.

The C/D snoRNAs represent an essential class of small nucleolar RNAs that guide 2'-O-ribose methylation of ribosomal RNAs and other RNAs in eukaryotes. In Arabidopsis thaliana more than a hundred C/D snoRNAs have been identified, most of them encoded by polycistronic gene clusters, but little is known on the factors controlling their biogenesis.

Here we focus on the identification of factors controlling the processing of tRNA-snoRNA dicistronic precursors (pre-tsnoRNA) synthesized by RNA polymerase III and producing tRNAGly and C/D snoR43. We produced radiolabelled RNA probes corresponding to different pre-tsnoRNA mutants to test their impact on processing in vitro by a recombinant tRNAse Z, the Arabidopsis endonuclease that processes the 3'end of tRNAs, and by nuclear extracts from cauliflower inflorescences that accurately process the pre-tsnoRNA. This was coupled to an in vivo analysis of the processing of tagged pre-tsnoRNA mutants expressed in Arabidopsis. Our results strongly implicate tRNase Z in endonucleolytic cleavage of the pre-tsnoRNA. In addition, they reveal an alternate pathway which could depend on a tRNA decay surveillance mechanism. Finally, we provide arguments showing that processing of pre-tsnoRNA, both in planta and by nuclear extracts, is coupled to the assembly of snoRNA with core proteins forming the functional snoRNP.




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G. Canino, E. Bocian, N. Barbezier, M. Echeverria, J. Forner, S. Binder, and A. Marchfelder
Arabidopsis Encodes Four tRNase Z Enzymes
Plant Physiology, July 1, 2009; 150(3): 1494 - 1502.
[Abstract] [Full Text] [PDF]


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