The final step of 40S ribosomal subunit maturation is controlled by a dual key lock

  1. Laura Plassart
  2. Ramtin Shayan
  3. Christian Montellese
  4. Dana Rinaldi
  5. Natacha Larburu
  6. Carole Pichereaux
  7. Carine Froment
  8. Simon Lebaron
  9. Marie-Françoise O'Donohue
  10. Ulrike Kutay
  11. Julien Marcoux
  12. Pierre-Emmanuel Gleizes  Is a corresponding author
  13. Celia Plisson-Chastang  Is a corresponding author
  1. Centre de Biologie Integrative, University of Toulouse, France
  2. ETH Zürich, Switzerland
  3. Institut de Pharmacologie et de Biologie Structurale, France
  4. CNRS, France

Abstract

Preventing premature interaction of pre-ribosomes with the translation apparatus is essential for translational accuracy. Hence, the final maturation step releasing functional 40S ribosomal subunits, namely processing of the 18S ribosomal RNA 3' end, is safeguarded by the protein DIM2, which both interacts with the endoribonuclease NOB1 and masks the rRNA cleavage site. To elucidate the control mechanism that unlocks NOB1 activity, we performed cryo-EM analysis of late human pre-40S particles purified using a catalytically-inactive form of the ATPase RIO1. These structures, together with in vivo and in vitro functional analyses, support a model in which ATP-loaded RIO1 cooperates with ribosomal protein RPS26/eS26 to displace DIM2 from the 18S rRNA 3' end, thereby triggering final cleavage by NOB1; release of ADP then leads to RIO1 dissociation from the 40S subunit. This dual key lock mechanism requiring RIO1 and RPS26 guarantees the precise timing of pre-40S particle conversion into translation-competent ribosomal subunits.

Data availability

Mass spectrometry proteomics data have been deposited to the ProteomeXchange Consortium via the PRIDE partner repository with the dataset identifier PXD019270. Cryo-EM maps have been deposited in the Electron Microscopy Data Bank (EMDB), under the accession codes : EMD-11440 (State A multi-body composite map); EMD-11441 (State B multi-body composite map); EMD-11446 (State A, head); EMD-11445 (State A, body); EMD-11447 (State A, platform); EMD-11443 (State B, head); EMD-11442 (State B, body); EMD-11444 (State B, platform). Atomic coordinate models of State A and State B RIO1(kd)-StHA pre-40S particles have been deposited in the Protein Data Bank (PDB), with respective PDB accession codes 6ZUO and 6ZV6.

Article and author information

Author details

  1. Laura Plassart

    Molecular, Cellular and Developmental biology department (MCD), Centre de Biologie Integrative, University of Toulouse, Toulouse, France
    Competing interests
    The authors declare that no competing interests exist.
  2. Ramtin Shayan

    Molecular, Cellular and Developmental biology department (MCD), Centre de Biologie Integrative, University of Toulouse, Toulouse, France
    Competing interests
    The authors declare that no competing interests exist.
  3. Christian Montellese

    Institute of Biochemistry, ETH Zürich, Zurich, Switzerland
    Competing interests
    The authors declare that no competing interests exist.
  4. Dana Rinaldi

    Molecular, Cellular and Developmental biology department (MCD), Centre de Biologie Integrative, University of Toulouse, Toulouse, France
    Competing interests
    The authors declare that no competing interests exist.
  5. Natacha Larburu

    Molecular, Cellular and Developmental biology department (MCD), Centre de Biologie Integrative, University of Toulouse, Toulouse, France
    Competing interests
    The authors declare that no competing interests exist.
  6. Carole Pichereaux

    Department of Biophysics, Institut de Pharmacologie et de Biologie Structurale, Toulouse, France
    Competing interests
    The authors declare that no competing interests exist.
  7. Carine Froment

    Institute of Pharmacology and Structural Biology, CNRS, Toulouse, France
    Competing interests
    The authors declare that no competing interests exist.
  8. Simon Lebaron

    Molecular, Cellular and Developmental biology department (MCD), Centre de Biologie Integrative, University of Toulouse, Toulouse, France
    Competing interests
    The authors declare that no competing interests exist.
  9. Marie-Françoise O'Donohue

    Molecular, Cellular and Developmental biology department (MCD), Centre de Biologie Integrative, University of Toulouse, Toulouse, France
    Competing interests
    The authors declare that no competing interests exist.
  10. Ulrike Kutay

    Institute of Biochemistry, ETH Zürich, Zurich, Switzerland
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-8257-7465
  11. Julien Marcoux

    Institute of Pharmacology and Structural Biology, CNRS, Toulouse, France
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-7321-7436
  12. Pierre-Emmanuel Gleizes

    Molecular, Cellular and Developmental biology department (MCD), Centre de Biologie Integrative, University of Toulouse, Toulouse, France
    For correspondence
    pierre-emmanuel.gleizes@univ-tlse3.fr
    Competing interests
    The authors declare that no competing interests exist.
  13. Celia Plisson-Chastang

    Molecular, Cellular and Developmental biology department (MCD), Centre de Biologie Integrative, University of Toulouse, Toulouse, France
    For correspondence
    celia.plisson-chastang@univ-tlse3.fr
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-8439-8428

Funding

Agence Nationale de la Recherche (16-CE11-0029)

  • Laura Plassart
  • Ramtin Shayan
  • Natacha Larburu
  • Simon Lebaron
  • Julien Marcoux
  • Pierre-Emmanuel Gleizes
  • Celia Plisson-Chastang

Swiss National Science Fundation (31003A_166565)

  • Christian Montellese

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Reviewing Editor

  1. Alan G Hinnebusch, Eunice Kennedy Shriver National Institute of Child Health and Human Development, United States

Publication history

  1. Received: July 20, 2020
  2. Accepted: April 19, 2021
  3. Accepted Manuscript published: April 28, 2021 (version 1)
  4. Version of Record published: May 11, 2021 (version 2)

Copyright

© 2021, Plassart et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

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  1. Laura Plassart
  2. Ramtin Shayan
  3. Christian Montellese
  4. Dana Rinaldi
  5. Natacha Larburu
  6. Carole Pichereaux
  7. Carine Froment
  8. Simon Lebaron
  9. Marie-Françoise O'Donohue
  10. Ulrike Kutay
  11. Julien Marcoux
  12. Pierre-Emmanuel Gleizes
  13. Celia Plisson-Chastang
(2021)
The final step of 40S ribosomal subunit maturation is controlled by a dual key lock
eLife 10:e61254.
https://doi.org/10.7554/eLife.61254

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