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Fluctuations between multiple EF-G-induced chimeric tRNA states during translocation on the ribosome

Author

Listed:
  • Sarah Adio

    (Max Planck Institute for Biophysical Chemistry)

  • Tamara Senyushkina

    (Max Planck Institute for Biophysical Chemistry)

  • Frank Peske

    (Max Planck Institute for Biophysical Chemistry)

  • Niels Fischer

    (3D Electron Cryomicroscopy Group, Max Planck Institute for Biophysical Chemistry)

  • Wolfgang Wintermeyer

    (Max Planck Institute for Biophysical Chemistry)

  • Marina V. Rodnina

    (Max Planck Institute for Biophysical Chemistry)

Abstract

The coupled translocation of transfer RNA and messenger RNA through the ribosome entails large-scale structural rearrangements, including step-wise movements of the tRNAs. Recent structural work has visualized intermediates of translocation induced by elongation factor G (EF-G) with tRNAs trapped in chimeric states with respect to 30S and 50S ribosomal subunits. The functional role of the chimeric states is not known. Here we follow the formation of translocation intermediates by single-molecule fluorescence resonance energy transfer. Using EF-G mutants, a non-hydrolysable GTP analogue, and fusidic acid, we interfere with either translocation or EF-G release from the ribosome and identify several rapidly interconverting chimeric tRNA states on the reaction pathway. EF-G engagement prevents backward transitions early in translocation and increases the fraction of ribosomes that rapidly fluctuate between hybrid, chimeric and posttranslocation states. Thus, the engagement of EF-G alters the energetics of translocation towards a flat energy landscape, thereby promoting forward tRNA movement.

Suggested Citation

  • Sarah Adio & Tamara Senyushkina & Frank Peske & Niels Fischer & Wolfgang Wintermeyer & Marina V. Rodnina, 2015. "Fluctuations between multiple EF-G-induced chimeric tRNA states during translocation on the ribosome," Nature Communications, Nature, vol. 6(1), pages 1-11, November.
    • Handle: RePEc:nat:natcom:v:6:y:2015:i:1:d:10.1038_ncomms8442
    DOI: 10.1038/ncomms8442
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    Cited by:

    1. Panagiotis Poulis & Anoshi Patel & Marina V. Rodnina & Sarah Adio, 2022. "Altered tRNA dynamics during translocation on slippery mRNA as determinant of spontaneous ribosome frameshifting," Nature Communications, Nature, vol. 13(1), pages 1-15, December.
    2. Sakshi Jain & Lukasz Koziej & Panagiotis Poulis & Igor Kaczmarczyk & Monika Gaik & Michal Rawski & Namit Ranjan & Sebastian Glatt & Marina V. Rodnina, 2023. "Modulation of translational decoding by m6A modification of mRNA," Nature Communications, Nature, vol. 14(1), pages 1-13, December.

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