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EF-G-dependent GTP hydrolysis induces translocation accompanied by large conformational changes in the 70S ribosome

Nature Structural Biology volume 6, pages 643–647 (1999)Cite this article

Abstract

Cryo-electron microscopy has been used to visualize elongation factor G (EF-G) on the 70S ribosome in GDP and GTP states. GTP hydrolysis is required for binding of all the domains of EF-G to the pretranslocational complex and for the completion of translocation. In addition, large conformational changes have been identified in the ribosome. The head of the 30S subunit shifts toward the L1 protein side, and the L7/L12 stalk becomes bifurcated upon EF-G binding. Upon GTP hydrolysis, the bifurcation is reversed and an arc-like connection is formed between the base of the stalk and EF-G.

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Figure 1: Three-dimensional cryo-EM maps.
Figure 2: Difference maps obtained by subtracting the three-dimensional map of the ribosome occupied by P-site tRNA11 from the three-dimensional maps of various EF-G–ribosome complexes overlaid on the 15 Å resolution map of the 70S ribosome11.
Figure 3: a, Stereo view comparison of masses of densities (wire-mesh contours) related to EF-G in the 70S–EF-G–GMPP(CH2)P complex (magenta) superimposed with the corresponding density obtained from the 70S–(tRNA)2–EF-G–GMPP(CH2)P complex (green), prepared using a higher concentration of EF-G.
Figure 4: Relative movement of the 30S subunit with respect to the 50S subunit upon EF-G–GMPP(CH2)P binding to the 70S ribosome.
Figure 5: Proposed sequence of events of the translocation reaction.

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Acknowledgements

We thank P. Moore for providing the all-atom coordinates of the EF-G–GDPX-ray structure. This work was supported by grants from the National Institutes of Health and the National Science Foundation.

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Authors and Affiliations

  1. Wadsworth Center, Empire State Plaza , Albany, 12201-0509, New York, USA

    Rajendra K. Agrawal, Amy B. Heagle, Pawel Penczek, Robert A. Grassucci & Joachim Frank

  2. Howard Hughes Medical Institute, Empire State Plaza, Albany, 12201-0509, New York, USA

    Amy B. Heagle, Robert A. Grassucci & Joachim Frank

  3. Department of Biomedical Sciences, State University of New York at Albany, Empire State Plaza, Albany, 12201-0509, New York, USA

    Pawel Penczek & Joachim Frank

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  1. Rajendra K. Agrawal
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Correspondence to Rajendra K. Agrawal.

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Agrawal, R., Heagle, A., Penczek, P. et al. EF-G-dependent GTP hydrolysis induces translocation accompanied by large conformational changes in the 70S ribosome. Nat Struct Mol Biol 6, 643–647 (1999). https://doi.org/10.1038/10695

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