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Originally published In Press as doi:10.1074/jbc.M211937200 on February 5, 2003

J. Biol. Chem., Vol. 278, Issue 15, 13026-13032, April 11, 2003
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Purification, Cloning, and Characterization of XendoU, a Novel Endoribonuclease Involved in Processing of Intron-encoded Small Nucleolar RNAs in Xenopus laevis*

Pietro LaneveDagger , Fabio Altieri§, Micol E. FioriDagger , Andrea Scaloni, Irene BozzoniDagger , and Elisa Caffarelli||**

From the Dagger  Institute Pasteur Fondazione Cenci-Bolognetti, Department of Genetics and Molecular Biology, University "La Sapienza" Piazzale Aldo Moro 5, 00185 Rome, Italy, the § Department of Biochemistry, University "La Sapienza," 00185 Rome, Italy,  Proteomics and Mass Spectrometry Laboratory, Institute for Animal Production System in Mediterranean Environment, National Research Council, 80147 Naples, Italy, and the || Institute of Molecular Biology and Pathology, National Research Council, 00185 Rome, Italy

Here we report the purification, from Xenopus laevis oocyte nuclear extracts, of a new endoribonuclease, XendoU, that is involved in the processing of the intron-encoded box C/D U16 small nucleolar RNA (snoRNA) from its host pre-mRNA. Such an activity has never been reported before and has several uncommon features that make it quite a novel enzyme: it is poly(U)-specific, it requires Mn2+ ions, and it produces molecules with 2'-3'-cyclic phosphate termini. Even if XendoU cleaves U-stretches, it displays some preferential cleavage on snoRNA precursor molecules. XendoU also participates in the biosynthesis of another intron-encoded snoRNA, U86, which is contained in the NOP56 gene of Xenopus laevis. A common feature of these snoRNAs is that their production is alternative to that of the mRNA, suggesting an important regulatory role for all the factors involved in the processing reaction.

* This work was supported in part by grants from Ministero dell'Università e delle Ricerca Scientifica e Tecnologica (Biotechnology Program L.95/95, Biotecnologie, Programmi di Ricerca di Interesse Nazionale 40%, and Centro di Eccellenza Biologia e Medicina Molecolare) and from Consiglio Nazionale delle Ricerche (Target Project on Biotechnology and Tecnologie di Base della Post-genomica).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/EBI Data Bank with accession number(s) AJ507315.

** To whom correspondence should be addressed: Inst. of Molecular Biology and Pathology, c/o Department of Genetics and Molecular Biology, University "La Sapienza" P. le Aldo Moro 5, 00185 Rome, Italy. Tel.: 39-06-4991-2217; Fax: 39-06-4991-2500; E-mail: elisa.caffarelli@uniroma1.it.

Copyright © 2003 by The American Society for Biochemistry and Molecular Biology, Inc.
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