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Originally published In Press as doi:10.1074/jbc.M002996200 on May 26, 2000

J. Biol. Chem., Vol. 275, Issue 35, 27212-27220, September 1, 2000
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Fibrillarin Genes Encode Both a Conserved Nucleolar Protein and a Novel Small Nucleolar RNA Involved in Ribosomal RNA Methylation in Arabidopsis thaliana*

Frédy Barneche, Florence Steinmetz, and Manuel EcheverríaDagger

From the Laboratoire Génome et Développement des Plantes, Université de Perpignan, Unité de Recherche Mixte CNRS 5096, 52 Avenue de Villeneuve, 66860 Perpignan Cedex, France

Fibrillarin is a key nucleolar protein in eukaryotes which associates with box C/D small nucleolar RNAs (snoRNAs) directing 2'-O-ribose methylation of the rRNA. In this study we describe two genes in Arabidopsis thaliana, AtFib1 and AtFib2, encoding nearly identical proteins conserved with eukaryotic fibrillarins. We demonstrate that AtFib1 and AtFib2 proteins are functional homologs of the yeast Nop1p (fibrillarin) and can rescue a yeast NOP1-null mutant strain. Surprisingly, for the first time in plants, we identified two isoforms of a novel box C/D snoRNA, U60.1f and U60.2f, nested in the fifth intron of AtFib1 and AtFib2. Interestingly after gene duplication the host intronic sequences completely diverged, but the snoRNA was conserved, even in other crucifer fibrillarin genes. We show that the U60f snoRNAs accumulate in seedlings and that their targeted residue on the 25 S rRNA is methylated. Our data reveal that the three modes of expression of snoRNAs, single, polycistronic, and intronic, exist in plants and suggest that the mechanisms directing rRNA methylation, dependent on fibrillarin and box C/D snoRNAs, are evolutionarily conserved in plants.

* This work was supported by a grant from the Ministère de l'Education Nationale, de l'Enseignement Superieur et de la Recherche (to F. B.).The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

The nucleotide sequence(s) reported in this paper has been submitted to the GenBankTM/EMBL Data Bank with accession number(s) AtFib1 gene, AF265547; AtFib2 gene, AF267171; AtFib1 cDNA, AF233443; AtFib2 cDNA, AF233444; U60.1f, AF228364; U60.2f; AF228365.

Dagger To whom correspondence should be addressed. Tel.: 334-6866 2132; Fax: 334-6866 8499; E-mail: manuel@univ-perp.fr.

Copyright © 2000 by The American Society for Biochemistry and Molecular Biology, Inc.
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