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Mol Cell Biol, February 1998, p. 1023-1028, Vol. 18, No. 2
0270-7306/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

In Vivo Identification of Nuclear Factors Interacting with the Conserved Elements of Box C/D Small Nucleolar RNAs

Elisa Caffarelli,1 Massimo Losito,2 Corinna Giorgi,2 Alessandro Fatica,2 and Irene Bozzoni2,*

Centro Acidi Nucleici of CNR1 and Dipartimento di Genetica e Biologia Molecolare, Istituto-Pasteur Fondazione Cenci-Bolognetti,2 Università "La Sapienza," Rome, Italy

Received 28 July 1997/Returned for modification 8 September 1997/Accepted 17 November 1997

The U16 small nucleolar RNA (snoRNA) is encoded by the third intron of the L1 (L4, according to the novel nomenclature) ribosomal protein gene of Xenopus laevis and originates from processing of the pre-mRNA in which it resides. The U16 snoRNA belongs to the box C/D snoRNA family, whose members are known to assemble in ribonucleoprotein particles (snoRNPs) containing the protein fibrillarin. We have utilized U16 snoRNA in order to characterize the factors that interact with the conserved elements common to the other members of the box C/D class. In this study, we have analyzed the in vivo assembly of U16 snoRNP particles in X. laevis oocytes and identified the proteins which interact with the RNA by label transfer after UV cross-linking. This analysis revealed two proteins, of 40- and 68-kDa apparent molecular size, which require intact boxes C and D together with the conserved 5',3'-terminal stem for binding. Immunoprecipitation experiments showed that the p40 protein corresponds to fibrillarin, indicating that this protein is intimately associated with the RNA. We propose that fibrillarin and p68 represent the RNA-binding factors common to box C/D snoRNPs and that both proteins are essential for the assembly of snoRNP particles and the stabilization of the snoRNA.

* Corresponding author. Mailing address: Istituto-Pasteur Fondazione Cenci-Bolognetti, Dipartimento di Genetica e Biologia Molecolare, Universita "La Sapienza," 00185 Rome, Italy. Phone: 39-6-49912202. Fax: 39-6-49912500. E-mail: Bozzoni{at}axcasp.caspur.it.




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