Structural aspects of messenger RNA reading frame maintenance by the ribosome
Abstract
One key question in protein biosynthesis is how the ribosome couples mRNA and tRNA movements to prevent disruption of weak codon-anticodon interactions and loss of the translational reading frame during translocation. Here we report the complete path of mRNA on the 70S ribosome at the atomic level (3.1-A resolution), and we show that one of the conformational rearrangements that occurs upon transition from initiation to elongation is a narrowing of the downstream mRNA tunnel. This rearrangement triggers formation of a network of interactions between the mRNA downstream of the A-site codon and the elongating ribosome. Our data elucidate the mechanism by which hypermodified nucleoside 2-methylthio-N6 isopentenyl adenosine at position 37 (ms(2)i(6)A37) in tRNA(Phe)(GAA) stabilizes mRNA-tRNA interactions in all three tRNA binding sites. Another network of contacts is formed between this tRNA modification and ribosomal elements surrounding the mRNA E/P kink, resulting in the anchoring of P-site tRNA. These data allow rationalization of how modification deficiencies of ms(2)i(6)A37 in tRNAs may lead to shifts of the translational reading frame.
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The structure of the 70S ribosome from Thermus thermophilus has >3000 modeled cations, all but 3 initially assigned as Mg2+.
“The structure of the 70S ribosome from Thermus thermophilus (PDB 4 V6F)51 has >3000 modeled cations (c). All but 3 of these were initially assigned as Mg2+.”
From Metals in Motion: Understanding Labile Metal Pools in Bacteria (2025) · cites this paper as [51]
>3000 cation-binding sites were visualized in the Thermus thermophilus 70S ribosome and assigned as Mg2+, but those assignments have been challenged on experimental and stereochemical grounds.
“As an upper limit, > 3000 cation-binding sites were visualized in the Thermus thermophilus 70S ribosome51 and assigned as Mg2+, but those assignments have been challenged on both experimental52 and stereochemical54 grounds.”
From Metals in Motion: Understanding Labile Metal Pools in Bacteria (2025) · cites this paper as [51]
Tetracycline binding at the second site has not been previously described.
“Tetracycline binding at this position has not been previously described (17 -20).”
From Resistance mutations generate divergent antibiotic susceptibility profiles against translation inhibitors (2016) · cites this paper as [19]
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- Authors
- Jenner, L. B., Demeshkina, N., Yusupova, G., & Yusupov, M.
- Affiliations
- Institut de génétique et de biologie moléculaire et cellulaire (FR)
- Centre National de la Recherche Scientifique (FR)
- Venue
- Nature Structural & Molecular Biology
- Published
- 2010-04-18
- Volume
- 17
- Issue
- 5
- Pages
- 555-560
- ISSN
- 1545-9985 (ISSN-L)
- 1545-9993
- OpenAlex
- W2073504319
- Cited by
- 312
- References
- 45
- Citations by year
- 2026: 6 · 2025: 9 · 2024: 13 · 2023: 16 · 2022: 14 · 2021: 10 · 2020: 22 · 2019: 16 · 2018: 22 · 2017: 21 · 2016: 26 · 2015: 23 · 2014: 28 · 2013: 30 · 2012: 28
- FWCI
- 11.20
- Type
- journal article
- Language
- en
- Open access
- closed
- Licence
- www.springer.com/tdm (tdm)
- Locations
- Nature Structural & Molecular Biology (publishedVersion)
- PubMed (publishedVersion)
- HAL (submittedVersion)
- DORA PSI (Paul Scherrer Institute) (submittedVersion)
- Funders
- Agence Nationale de la Recherche (ANR-07-PCVI-0015)
- 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 › Molecular Biology › DNA and Nucleic Acid Chemistry
- Keywords
- ribosome, codon-anticodon pairing, 70S ribosome, tRNA modification, tRNA-Phe, ribosome structure, translational readthrough
- MeSH terms
- Escherichia coli, Models, Molecular, Nucleic Acid Conformation, RNA, Bacterial, RNA, Messenger, RNA, Transfer, Reading Frames, Thermus thermophilus, Crystallography, X-Ray, Ribosome Subunits
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