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Molecular and Cellular Biology, July 2003, p. 4449-4460, Vol. 23, No. 13
0270-7306/03/$08.00+0 DOI: 10.1128/MCB.23.13.4449-4460.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.
Sequential Protein Association with Nascent 60S Ribosomal Particles
Cosmin Saveanu,1 Abdelkader Namane,2 Pierre-Emmanuel Gleizes,3 Alice Lebreton,1 Jean-Claude Rousselle,2 Jacqueline Noaillac-Depeyre,3 Nicole Gas,3 Alain Jacquier,1 and Micheline Fromont-Racine1*
Génétique des Interactions Macromoléculaires, CNRS-URA2171,1
PT Protéomique, Institut Pasteur, 75724 Paris Cedex 15,2
Laboratoire de Biologie Moléculaire Eucaryote (CNRS), 31062 Toulouse Cedex, France3
Received 22 January 2003/
Returned for modification 28 February 2003/
Accepted 4 April 2003
Ribosome biogenesis in eukaryotes depends on the coordinated action of ribosomal and nonribosomal proteins that guide the assembly of preribosomal particles. These intermediate particles follow a maturation pathway in which important changes in their protein composition occur. The mechanisms involved in the coordinated assembly of the ribosomal particles are poorly understood. We show here that the association of preribosomal factors with pre-60S complexes depends on the presence of earlier factors, a phenomenon essential for ribosome biogenesis. The analysis of the composition of purified preribosomal complexes blocked in maturation at specific steps allowed us to propose a model of sequential protein association with, and dissociation from, early pre-60S complexes for several preribosomal factors such as Mak11, Ssf1, Rlp24, Nog1, and Nog2. The presence of either Ssf1 or Nog2 in complexes that contain the 27SB pre-rRNA defines novel, distinct pre-60S particles that contain the same pre-rRNA intermediates and that differ only by the presence or absence of specific proteins. Physical and functional interactions between Rlp24 and Nog1 revealed that the assembly steps are, at least in part, mediated by direct protein-protein interactions.
* Corresponding author. Mailing address: Génétique des Interactions Macromoléculaires, Institut Pasteur (CNRS-URA2171), 25-28 rue du Dr. Roux, 75724 Paris Cedex 15, France. Phone: 33 0 1 40 61 32 05. Fax: 33 0 1 45 68 87 90. E-mail: mfromont{at}pasteur.fr.
Molecular and Cellular Biology, July 2003, p. 4449-4460, Vol. 23, No. 13
0022-538X/03/$08.00+0 DOI: 10.1128/MCB.23.13.4449-4460.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.
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Copyright © 2003 by the American Society for Microbiology. All rights reserved.