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Assembly and functionality of the ribosome with tethered subunits

Author

Listed:
  • Nikolay A. Aleksashin

    (University of Illinois at Chicago)

  • Margus Leppik

    (University of Tartu)

  • Adam J. Hockenberry

    (Northwestern University
    The University of Texas at Austin)

  • Dorota Klepacki

    (University of Illinois at Chicago)

  • Nora Vázquez-Laslop

    (University of Illinois at Chicago)

  • Michael C. Jewett

    (Northwestern University)

  • Jaanus Remme

    (University of Tartu)

  • Alexander S. Mankin

    (University of Illinois at Chicago)

Abstract

Ribo-T is an engineered ribosome whose small and large subunits are tethered together by linking 16S rRNA and 23S rRNA in a single molecule. Although Ribo-T can support cell proliferation in the absence of wild type ribosomes, Ribo-T cells grow slower than those with wild type ribosomes. Here, we show that cell growth defect is likely explained primarily by slow Ribo-T assembly rather than its imperfect functionality. Ribo-T maturation is stalled at a late assembly stage. Several post-transcriptional rRNA modifications and some ribosomal proteins are underrepresented in the accumulated assembly intermediates and rRNA ends are incompletely trimmed. Ribosome profiling of Ribo-T cells shows no defects in translation elongation but reveals somewhat higher occupancy by Ribo-T of the start codons and to a lesser extent stop codons, suggesting that subunit tethering mildly affects the initiation and termination stages of translation. Understanding limitations of Ribo-T system offers ways for its future development.

Suggested Citation

  • Nikolay A. Aleksashin & Margus Leppik & Adam J. Hockenberry & Dorota Klepacki & Nora Vázquez-Laslop & Michael C. Jewett & Jaanus Remme & Alexander S. Mankin, 2019. "Assembly and functionality of the ribosome with tethered subunits," Nature Communications, Nature, vol. 10(1), pages 1-13, December.
    • Handle: RePEc:nat:natcom:v:10:y:2019:i:1:d:10.1038_s41467-019-08892-w
    DOI: 10.1038/s41467-019-08892-w
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    Cited by:

    1. Yuishin Kosaka & Yumi Miyawaki & Megumi Mori & Shunsuke Aburaya & Chisato Nishizawa & Takeshi Chujo & Tatsuya Niwa & Takumi Miyazaki & Takashi Sugita & Mao Fukuyama & Hideki Taguchi & Kazuhito Tomizaw, 2025. "Autonomous ribosome biogenesis in vitro," Nature Communications, Nature, vol. 16(1), pages 1-14, December.

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