A large nucleolar U3 ribonucleoprotein required for 18S ribosomal RNA biogenesis

Abstract

Although the U3 small nucleolar RNA (snoRNA), a member of the box C/D class of snoRNAs, was identified with the spliceosomal small nuclear RNAs (snRNAs) over 30 years ago1,2, its function and its associated protein components have remained more elusive. The U3 snoRNA is ubiquitous in eukaryotes and is required for nucleolar processing of pre-18S ribosomal RNA in all organisms where it has been tested3,4. Biochemical and genetic analyses suggest that U3–pre-rRNA base-pairing interactions mediate endonucleolytic pre-rRNA cleavages3. Here we have purified a large ribonucleoprotein (RNP) complex from Saccharomyces cerevisiae that contains the U3 snoRNA and 28 proteins. Seventeen new proteins (Utp1–17) and Rrp5 were present, as were ten known components. The Utp proteins are nucleolar and specifically associated with the U3 snoRNA. Depletion of the Utp proteins impedes production of the 18S rRNA, indicating that they are part of the active pre-rRNA processing complex. On the basis of its large size (80S; calculated relative molecular mass of at least 2,200,000) and function, this complex may correspond to the terminal knobs present at the 5′ ends of nascent pre-rRNAs. We have termed this large RNP the small subunit (SSU) processome.

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Figure 1: RNA composition of fractions from the purification of the U3 snoRNP.
Figure 2: Function of the new components of the SSU processome.
Figure 3: The SSU processome sediments at 80S on sucrose gradients.
Figure 4: The U3 snoRNA and Utp7 are required for terminal knob formation on nascent pre-rRNAs.

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Acknowledgements

We thank A. Djikeng, G. Dreyfuss, P. Gordon, J. Laney, T. Serio and T. Stone for assistance and advice. We also thank P. Glazer, M. Snyder and J. Steitz for critical reading of the manuscript. F.D. was supported by the Anna Fuller Fund for Molecular Oncology. This work was supported by federal grants from the NIH to D.F.H and S.J.B. and the NSF to A.L.B., and by The Patterson Trust to S.J.B.

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Correspondence to Susan J. Baserga.

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Dragon, F., Gallagher, J., Compagnone-Post, P. et al. A large nucleolar U3 ribonucleoprotein required for 18S ribosomal RNA biogenesis. Nature 417, 967–970 (2002). https://doi.org/10.1038/nature00769

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