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Molecular and Cellular Biology, February 2007, p. 1207-1221, Vol. 27, No. 4
0270-7306/07/$08.00+0     doi:10.1128/MCB.01523-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Characterization of Saccharomyces cerevisiae Npa2p (Urb2p) Reveals a Low-Molecular-Mass Complex Containing Dbp6p, Npa1p (Urb1p), Nop8p, and Rsa3p Involved in Early Steps of 60S Ribosomal Subunit Biogenesis{triangledown} ,{dagger}

Iván V. Rosado,1 Christophe Dez,2 Simon Lebaron,2 Michèle Caizergues-Ferrer,2 Yves Henry,2 and Jesús de la Cruz1*

Departamento de Genética, Universidad de Sevilla, Sevilla, Spain,1 Equipe Labellisée Ligue Nationale contre le Cancer, Laboratoire de Biologie Moleculaire Eucaryote, CNRS-Universite Paul Sabatier, IFR 109, Toulouse, France2

Received 16 August 2006/ Returned for modification 8 October 2006/ Accepted 24 November 2006

We report the characterization of the yeast Npa2p (Urb2p) protein, which is essential for 60S ribosomal subunit biogenesis. We identified this protein in a synthetic lethal screening with the rsa3 null allele. Rsa3p is a genetic partner of the putative RNA helicase Dbp6p. Mutation or depletion of Npa2p leads to a net deficit in 60S subunits and a decrease in the levels all 27S pre-rRNAs and mature 25S and 5.8S rRNAs. This is likely due to instability of early pre-60S particles. Consistent with a role of Npa2p in 60S subunit biogenesis, green fluorescent protein-tagged Npa2p localizes predominantly to the nucleolus and TAP-tagged Npa2p sediments with large complexes in sucrose gradients and is associated mainly with 27SA2 pre-rRNA-containing preribosomal particles. In addition, we reveal a genetic synthetic interaction between Npa2p, several factors required for early steps of 60S subunit biogenesis (Dbp6p, Dbp7p, Dbp9p, Npa1p, Nop8p, and Rsa3p), and the 60S protein Rpl3p. Furthermore, coimmunoprecipitation and gel filtration analyses demonstrated that at least Npa2p, Dbp6p, Npa1p, Nop8p, and Rsa3p are present together in a subcomplex of low molecular mass whose integrity is independent of RNA. Our results support the idea that these five factors work in concert during the early steps of 60S subunit biogenesis.

* Corresponding author. Mailing address: Departamento de Genética, Facultad de Biología, Universidad de Sevilla, Avda. Reina Mercedes, 6, E-41012 Sevilla, Spain. Phone: 34 95 455 71 06. Fax: 34 95 455 71 04. E-mail: jdlcd{at}us.es.

{triangledown} Published ahead of print on 4 December 2006.

{dagger} Supplemental material for this article may be found at http://mcb.asm.org/.

Molecular and Cellular Biology, February 2007, p. 1207-1221, Vol. 27, No. 4
0270-7306/07/$08.00+0     doi:10.1128/MCB.01523-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



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