A chaperone for ribosome maturation.
The nascent pre-rRNA of eukaryotic ribosomes is fully transcribed and assembled into an 80-90 S nucleolar particle before being cleaved into mature ribosomal RNA. The interdependence of steps in the processing of this precursor RNA indicates that RNA processing, at least in part, acts as a quality control mechanism that helps ensure that only functional RNA is incorporated into mature ribosomes. In search of structural components that underlie this interdependence using the Schizosaccharomyces pombe internal transcribed spacer 1 (ITS) as a ligand for affinity chromatography of ITS1-specific proteins, we have isolated a large spliceosome-like protein complex, a ribosome assembly chaperone (RAC) of 20 or more polypeptides (Lalev, A. I., Abeyrathne, P. D., and Nazar, R. N. (2000) J. Mol. Biol. 302, 65-77). When the ITS2 spacer was used in the present study to isolate ITS2-specific proteins, the same proteins were identified consistent with a complex containing multiple specific binding sites. Subsequent competition binding studies indicated that the protein complex actually contains independent binding sites for all four of the transcribed spacers in the pre-rRNA. Because disruption of protein-binding sites in these spacer RNAs is known to severely affect rRNA processing, taken together these results suggest that the RAC complex is a chaperone for ribosome maturation acting as a "rack" on which critical structure is organized.
Department of Molecular Biology and Genetics, University of Guelph, Guelph, Ontario N1G 2W1, Canada.
A I
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Lalev
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Nazar
D001665:Binding Sites; D002466:Cell Nucleolus / Q000502:physiology / Q000648:ultrastructure; D004271:DNA, Fungal / Q000235:genetics; D021903:DNA, Ribosomal Spacer / Q000235:genetics*; D005656:Fungal Proteins / Q000302:isolation & purification / Q000378:metabolism; D008961:Models, Structural; D012322:RNA Precursors / Q000235:genetics*; D012331:RNA, Fungal / Q000235:genetics; D012339:RNA, Ribosomal, 28S / Q000235:genetics / Q000378:metabolism; D012261:Ribonucleoproteins / Q000302:isolation & purification / Q000378:metabolism*; D012269:Ribosomal Proteins / Q000737:chemistry / Q000378:metabolism; D012270:Ribosomes / Q000235:genetics / Q000502:physiology* / Q000648:ultrastructure; D012568:Schizosaccharomyces / Q000235:genetics*; D014158:Transcription, Genetic
D004271:DNA, Fungal; D021903:DNA, Ribosomal Spacer; D005656:Fungal Proteins; D012322:RNA Precursors; D012331:RNA, Fungal; D012339:RNA, Ribosomal, 28S; D012261:Ribonucleoproteins; D012269:Ribosomal Proteins