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Visualization of elongation factor Tu on the Escherichia coli ribosome

Nature volume 389, pages 403–406 (1997)Cite this article

Abstract

The delivery of a specific amino acid to the translating ribosome is fundamental to protein synthesis. The binding of aminoacyl-transfer RNA to the ribosome is catalysed by the elongation factor Tu (EF-Tu). The elongation factor, the aminoacyl-tRNA and GTP form a stable ‘ternary’ complex that binds to the ribosome. We have used electron cryomicroscopy and angular reconstitution1 to visualize directly the kirromycin-stalled ternary complex in the A site of the 70S ribosome of Escherichia coli. Electron cryomicroscopy had previously given detailed ribosomal structures at 25 and 23 Å (refs 2, 3) resolution, and was used to determine the position of tRNAs on the ribosome4,5. In particular, the structures5 of pre-translocational (tRNAs in A and P sites) and post-translocational ribosomes (P and E sites occupied) were both visualized at a resolution of ∼20 Å. Our three-dimensional reconstruction at 18 Å resolution shows the ternary complex spanning the inter-subunit space with the acceptor domain of the tRNA reaching into the decoding centre. Domain 1 (the G domain) of the EF-Tu is bound both to the L7/L12 stalk and to the 50S body underneath the stalk, whereas domain 2 is oriented towards the S12 region on the 30S subunit.

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Figure 1: a, b, The 70S E.coli ribosome with the ternary complex locked into the A site by kirromycin.
Figure 2: Stereo views of the ternary complex on the 70S E.coli ribosome seen in directions ∼40° apart, all with the 30S subunit on the left and the 50S subunit on the right.
Figure 3: The crystallographic structure of the ternary complex (at 2.7 Å resolution) compared to the electron microscopical reconstruction (18 Å resolution).

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Acknowledgements

We thank J. Nyborg for the X-ray coordinates of the ternary complex. We acknowledge support with the IMAGIC system from M. Schatz and R. Schmidt of Image Science GmbH. The work was supported by the Deutsche Forschungsgemeinschaft, grants to W.W., M.vH. and R.B.; H.S. was financed in part by an award to R. Henderson of the MRC Laboratory for Molecular Biology, Cambridge, UK.

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Authors and Affiliations

  1. Department of Biochemistry, Imperial College of Science, Medicine and Technology, London, SW7 2AY, UK

    Holger Stark & Marin van Heel

  2. Fritz-Haber-Institut der Max-Planck-Gesellschaft, Berlin, D-14195, Germany

    Holger Stark

  3. Institut fr Molekularbiologie, Universitt Witten/Herdecke, D-58448, Witten, Germany

    Marina V. Rodnina & Wolfgang Wintermeyer

  4. Max-Planck-Institut fr Molekulare Genetik, D-14195, Berlin, Germany

    Jutta Rinke-Appel & Richard Brimacombe

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  1. Holger Stark
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Correspondence to Marin van Heel.

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Stark, H., Rodnina, M., Rinke-Appel, J. et al. Visualization of elongation factor Tu on the Escherichia coli ribosome. Nature 389, 403–406 (1997). https://doi.org/10.1038/38770

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