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© The Rockefeller University Press, 0021-9525/1999//1369 $5.00
The Journal of Cell Biology, Volume 145, Number 7, , 1999 1369-1380

Regular Articles

Assembly of 5S Ribosomal RNA Is Required at a Specific Step of the Pre-rRNA Processing Pathway



Anne-Marie Dechampesme*, Olga Koroleva*, Isabelle Leger-Silvestre{ddagger}, Nicole Gas{ddagger}, and Sylvie Camier*

* Service de Biochimie et de Génétique Moléculaire, Commissariat á L'Energie Atomique (CEA)/Saclay, F-91191 Gif-sur-Yvette Cedex, France; and {ddagger} Laboratoire de Biologie Moléculaire des Eucaryotes du Centre National de la Recherche Scientifique, F-31062 Toulouse Cedex, France

A collection of yeast strains surviving with mutant 5S RNA has been constructed. The mutant strains presented alterations of the nucleolar structure, with less granular component, and a delocalization of the 25S rRNA throughout the nucleoplasm. The 5S RNA mutations affected helix I and resulted in decreased amounts of stable 5S RNA and of the ribosomal 60S subunits. The shortage of 60S subunits was due to a specific defect in the processing of the 27SB precursor RNA that gives rise to the mature 25S and 5.8S rRNA. The processing rate of the 27SB pre-rRNA was specifically delayed, whereas the 27SA and 20S pre-rRNA were processed at a normal rate. The defect was partially corrected by increasing the amount of mutant 5S RNA. We propose that the 5S RNA is recruited by the pre-60S particle and that its recruitment is necessary for the efficient processing of the 27SB RNA precursor. Such a mechanism could ensure that all newly formed mature 60S subunits contain stoichiometric amounts of the three rRNA components.

Key Words: 5S ribosomal RNA • pre-rRNA • ribosomes • yeasts • cell nucleolus

Abbreviations used in this paper: ITS2, internal transcribed spacer 2; YPD, yeast extract/peptone/glucose..

We are very thankful to André Sentenac (CEA/Saclay, France) for his critical review of the manuscript and his constant support during this work. We also thank Pierre Thuriaux (CEA/Saclay, France) for his comments and interest in the study. We are very grateful to Françoise Wyers from François Lacroute's lab (Centre de Génétique Moléculaire, Centre de la Recherche Scientifique, Gif-sur-Yvette, France) for her help in the set up of the analysis of the ribosomal profiles. We also thank John Woolford, Jacqueline Segall (University of Toronto, Canada), Stuart Clarkson, Rosalia Arrebola (CEA/Saclay, France), and John Aris for their gifts of plasmids, yeast strains, and antibodies.


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