The final step of 40S ribosomal subunit maturation is controlled by a dual key lock
- Centre de Biologie Integrative, University of Toulouse, France
- ETH Zürich, Switzerland
- Institut de Pharmacologie et de Biologie Structurale, France
- 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
Celia Plisson-Chastang
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
- Alan G Hinnebusch, Eunice Kennedy Shriver National Institute of Child Health and Human Development, United States
Version history
- Received: July 20, 2020
- Accepted: April 19, 2021
- Accepted Manuscript published: April 28, 2021 (version 1)
- 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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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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