Role of Domains 4 and 5 in Elongation Factor G Functions on the Ribosome
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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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).”
From Ribosomal Protection Proteins and Their Mechanism ofTetracyclineResistance (2003) · cites this paper as [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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