Cited by 115RNA and protein synthesis mechanismsBacterial Genetics and BiotechnologyRNA modifications and cancer

Role of Domains 4 and 5 in Elongation Factor G Functions on the Ribosome

Savelsbergh, Andreas, Matassova, Natalia B., Rodnina, Marina V., Wintermeyer, Wolfgang·2000·Journal of Molecular Biology

Abstract

Elongation factor G (EF-G) is a large, five domain GTPase that catalyses the translocation of the tRNAs on the bacterial ribosome at the expense of GTP. In the crystal structure of GDP-bound EF-G, domain 1 (G domain) makes direct contacts with domains 2 and 5, whereas domain 4 protrudes from the body of the molecule. Here, we show that the presence of both domains 4 and 5 is essential for tRNA translocation and for the turnover of the factor on the ribosome, but not for rapid single-round GTP hydrolysis by EF-G. Replacement of a highly conserved histidine residue at the tip of domain 4, His583, with lysine or arginine decreases the rate of tRNA translocation at least 100-fold, whereas the binding of the factor to the ribosome, GTP hydrolysis and P(i) release are not affected by the mutations. Various small deletions in the tip region of domain 4 decrease the translocation activity of EF-G even further, but do not block the turnover of the factor. Unlike native EF-G, the mutants of EF-G lacking domains 4/5 do not interact with the alpha-sarcin stem-loop of 23 S rRNA. These mutants are not released from the ribosome after GTP hydrolysis or translocation, indicating that the contact with, or a conformational change of, the alpha-sarcin stem-loop is required for EF-G release from the ribosome.

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Registry

Crossref · 10.1006/jmbi.2000.3886

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  • Domain IV in EF-G has been implicated as an important determinant for promoting translocation of the tRNAs.

    “This is significant, as domain IV in EF-G has been implicated as an important determinant for promoting translocation of the tRNAs (30, 31, 42).”

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Authors
Savelsbergh, A., Matassova, N. B., Rodnina, M. V., & Wintermeyer, W.
Affiliations
  • Witten/Herdecke University (DE)
  • Max Planck Society (DE)
Venue
Journal of Molecular Biology
Publisher
Elsevier BV
Published
2000-07
Volume
300
Issue
4
Pages
951-961
ISSN
0022-2836 (ISSN-L)
OpenAlex
W1984294833
Cited by
115
References
45
Citations by year
2025: 2 · 2023: 4 · 2022: 7 · 2021: 5 · 2020: 10 · 2019: 7 · 2018: 2 · 2017: 1 · 2016: 2 · 2015: 4 · 2014: 5 · 2013: 5 · 2012: 4
FWCI
2.41
Type
journal article
Language
en
Open access
closed
Locations
  • Journal of Molecular Biology (publishedVersion)
  • PubMed (publishedVersion)
  • Max Planck Institute for Plasma Physics (submittedVersion)
  • GRO (submittedVersion)
  • MPG.PuRe (Max Planck Society) (submittedVersion)
Funders
Deutsche Forschungsgemeinschaft
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA and protein synthesis mechanisms
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Bacterial Genetics and Biotechnology
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA modifications and cancer
Keywords
elongation factor G, tRNA translocation, ribosome, GTPase, GTP hydrolysis, 23S rRNA, bacterial translation
MeSH terms
Binding Sites, Catalysis, Endoribonucleases, Escherichia coli, Fungal Proteins, Guanosine Triphosphate, Hydrolysis, Kinetics, Models, Molecular, Nucleic Acid Conformation, Ribosomes, RNA, Ribosomal, 23S, RNA, Transfer, Sulfuric Acid Esters, RNA-Binding Proteins, Conserved Sequence, Sequence Deletion, Protein Structure, Tertiary, Crystallography, X-Ray, Amino Acid Substitution, Peptide Elongation Factor G

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