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Ribosome engineering reveals the importance of 5S rRNA autonomy for ribosome assembly

Author

Listed:
  • Shijie Huang

    (University of Illinois at Chicago
    University of Pennsylvania)

  • Nikolay A. Aleksashin

    (University of Illinois at Chicago
    University of Illinois at Chicago
    University of California)

  • Anna B. Loveland

    (RNA Therapeutics Institute, Department of Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School)

  • Dorota Klepacki

    (University of Illinois at Chicago)

  • Kaspar Reier

    (Institute of Molecular and Cellular Biology, University of Tartu)

  • Amira Kefi

    (University of Illinois at Chicago)

  • Teresa Szal

    (University of Illinois at Chicago)

  • Jaanus Remme

    (Institute of Molecular and Cellular Biology, University of Tartu)

  • Luc Jaeger

    (Chemistry and Biochemistry Department, University of California)

  • Nora Vázquez-Laslop

    (University of Illinois at Chicago
    University of Illinois at Chicago)

  • Andrei A. Korostelev

    (RNA Therapeutics Institute, Department of Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School)

  • Alexander S. Mankin

    (University of Illinois at Chicago
    University of Illinois at Chicago)

Abstract

5S rRNA is an indispensable component of cytoplasmic ribosomes in all species. The functions of 5S rRNA and the reasons for its evolutionary preservation as an independent molecule remain unclear. Here we used ribosome engineering to investigate whether 5S rRNA autonomy is critical for ribosome function and cell survival. By linking circularly permutated 5S rRNA with 23S rRNA we generated a bacterial strain devoid of free 5S rRNA. Viability of the engineered cells demonstrates that autonomous 5S rRNA is dispensable for cell growth under standard conditions and is unlikely to have essential functions outside the ribosome. The fully assembled ribosomes carrying 23S-5S rRNA are highly active in translation. However, the engineered cells accumulate aberrant 50S subunits unable to form stable 70S ribosomes. Cryo-EM analysis revealed a malformed peptidyl transferase center in the misassembled 50S subunits. Our results argue that the autonomy of 5S rRNA is preserved due to its role in ribosome biogenesis.

Suggested Citation

  • Shijie Huang & Nikolay A. Aleksashin & Anna B. Loveland & Dorota Klepacki & Kaspar Reier & Amira Kefi & Teresa Szal & Jaanus Remme & Luc Jaeger & Nora Vázquez-Laslop & Andrei A. Korostelev & Alexander, 2020. "Ribosome engineering reveals the importance of 5S rRNA autonomy for ribosome assembly," Nature Communications, Nature, vol. 11(1), pages 1-13, December.
    • Handle: RePEc:nat:natcom:v:11:y:2020:i:1:d:10.1038_s41467-020-16694-8
    DOI: 10.1038/s41467-020-16694-8
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