Structures of modified eEF2·80S ribosome complexes reveal the role of GTP hydrolysis in translocation
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On the basis of kinetic data on ribosome protein synthesis, the mechanical energy for translocation of the mRNA-tRNA complex is thought to be provided by GTP hydrolysis of an elongation factor (eEF2 in eukaryotes, EF-G in bacteria). We have obtained cryo-EM reconstructions of eukaryotic ribosomes complexed with ADP-ribosylated eEF2 (ADPR-eEF2), before and after GTP hydrolysis, providing a structural basis for analyzing the GTPase-coupled mechanism of translocation. Using the ADP-ribosyl group as a distinct marker, we observe conformational changes of ADPR-eEF2 that are due strictly to GTP hydrolysis. These movements are likely representative of native eEF2 motions in a physiological context and are sufficient to uncouple the mRNA-tRNA complex from two universally conserved bases in the ribosomal decoding center (A1492 and A1493 in Escherichia coli) during translocation. Interpretation of these data provides a detailed two-step model of translocation that begins with the eEF2/EF-G binding-induced ratcheting motion of the small ribosomal subunit. GTP hydrolysis then uncouples the mRNA-tRNA complex from the decoding center so translocation of the mRNA-tRNA moiety may be completed by a head rotation of the small subunit.
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GTP hydrolysis induces substantial conformational changes in the ribosome.
“It is known that GTP hydrolysis induces substantial conformational changes in the ribosome.”
From Mechanism of tetracycline resistance by ribosomal protection protein Tet(O) (2013) · cites this paper as [25]
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- Authors
- Taylor, D. J., Nilsson, J., Merrill, A. R., Andersen, G. R., Nissen, P., & Frank, J.
- Affiliations
- Wadsworth Center (US)
- Aarhus University (DK)
- University of Guelph (CA)
- Corresponding author
- Frank, Joachim
- Venue
- The EMBO Journal
- Published
- 2007-04-19
- Volume
- 26
- Issue
- 9
- Pages
- 2421-2431
- ISSN
- 0261-4189 (ISSN-L)
- 1460-2075
- PMC
- PMC1864975
- OpenAlex
- W2086666821
- Cited by
- 193
- References
- 59
- Citations by year
- 2026: 4 · 2025: 2 · 2024: 4 · 2023: 12 · 2022: 11 · 2021: 8 · 2020: 2 · 2019: 5 · 2018: 14 · 2017: 6 · 2016: 11 · 2015: 4 · 2014: 7 · 2013: 18 · 2012: 20
- FWCI
- 7.03
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Licence
- Locations
- The EMBO Journal (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- Funders
- Howard Hughes Medical Institute
- Novo Nordisk
- Novo Nordisk Fonden
- Canadian Institutes of Health Research
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA and protein synthesis mechanisms
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA modifications and cancer
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Structural Biology › Advanced Electron Microscopy Techniques and Applications
- Keywords
- elongation factor 2, GTP hydrolysis, ribosome translocation, ADP-ribosylation, elongation factors, 80S ribosome, head rotation, elongation factor G
- MeSH terms
- Adenosine Diphosphate, Ascomycota, Guanosine Triphosphate, Hydrolysis, Models, Molecular, Molecular Sequence Data, Ribosomes, RNA, Messenger, RNA, Transfer, Protein Structure, Tertiary, Cryoelectron Microscopy, Peptide Elongation Factor 2, Peptide Elongation Factor G, RNA Transport
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