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Stage-specific assembly events of the 6-MDa small-subunit processome initiate eukaryotic ribosome biogenesis

Nature Structural & Molecular Biology volume 22, pages 920923 (2015) | Download Citation

Abstract

Eukaryotic ribosome biogenesis involves a plethora of ribosome-assembly factors, and their temporal order of association with preribosomal RNA is largely unknown. By using Saccharomyces cerevisiae as a model organism, we developed a system that recapitulates and arrests ribosome assembly at early stages, thus providing in vivo snapshots of nascent preribosomal particles. Here we report the stage-specific order in which 70 ribosome-assembly factors associate with preribosomal RNA domains, thereby forming the 6-MDa small-subunit processome.

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Acknowledgements

We thank Z. Hakhverdyan, J. Fernandez-Martinez and M. Rout for early advice with nanobody affinity purification, S. Gerstberger for technical assistance with northern blots and S. Liebman (University of Reno, Nevada) for the kind gift of yeast strains L-1491 and L-1496. M.C-M. is supported by a scholarship from Fonds de Recherche du Québec–Santé (FRQ-S). J.B. is supported by an European Molecular Biology Organization long-term fellowship (ALTF 51-2014). S.K. is supported by the Robertson Foundation, the Alfred P. Sloan Foundation, the Irma T. Hirschl Trust and the Human Frontier Science Program. The Proteomics Resource Center at Rockefeller University acknowledges funding from the Leona M. and Harry B. Helmsley Charitable Trust for mass spectrometer instrumentation.

Author information

Affiliations

  1. Laboratory of Protein and Nucleic Acid Chemistry, Rockefeller University, New York, New York, USA.

    • Malik Chaker-Margot
    • , Mirjam Hunziker
    • , Jonas Barandun
    •  & Sebastian Klinge
  2. Tri-Institutional Training Program in Chemical Biology, Rockefeller University, New York, New York, USA.

    • Malik Chaker-Margot
  3. Proteomics Resource Center, Rockefeller University, New York, New York, USA.

    • Brian D Dill

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Contributions

M.C.-M. and S.K. conceived the project. M.C.-M. performed all biochemical experiments. M.H. and J.B. established RNP protein-purification procedures. B.D.D. performed mass spectrometry analysis. M.C.-M. and S.K. analyzed the results and wrote the manuscript. All authors edited the manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Sebastian Klinge.

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DOI

https://doi.org/10.1038/nsmb.3111

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