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Doublet decoding of tRNASer3 demonstrates plasticity of ribosomal decoding center

Krishnaswamy, Shruthi ; Akbar, Shirin ; Larsson, Daniel S.D. ; Chen, Yang LU and Selmer, Maria (2025) In Nature Communications 16(1).
Abstract

Frameshifts can be caused by specific combinations of tRNA and mRNA. The wildtype AGC-decoding E. coli tRNASer3GCU has been shown to induce −1 ribosomal frameshifting on GCA alanine codons, and proposed to read a two-base codon instead of a canonical triplet. However, it has remained unclear whether this type of non-cognate decoding can be accommodated by the ribosome. Here, we perform single-particle cryo-EM reconstructions on E. coli 70S ribosomes with the frameshift-inducing tRNASer3 bound to the non-cognate GCA codon or the cognate AGC codon in the ribosomal A site. The structures demonstrate that doublet decoding is made possible when A1493, the conserved monitoring base in 16S rRNA, mimics a first... (More)

Frameshifts can be caused by specific combinations of tRNA and mRNA. The wildtype AGC-decoding E. coli tRNASer3GCU has been shown to induce −1 ribosomal frameshifting on GCA alanine codons, and proposed to read a two-base codon instead of a canonical triplet. However, it has remained unclear whether this type of non-cognate decoding can be accommodated by the ribosome. Here, we perform single-particle cryo-EM reconstructions on E. coli 70S ribosomes with the frameshift-inducing tRNASer3 bound to the non-cognate GCA codon or the cognate AGC codon in the ribosomal A site. The structures demonstrate that doublet decoding is made possible when A1493, the conserved monitoring base in 16S rRNA, mimics a first codon base, forming a Hoogsteen base pair with U36 from the anticodon and stacking with the mRNA. This interaction pushes the first two bases of the A-site codon in position for base pairing with C35 and G34 of the anticodon.

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author
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organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Nature Communications
volume
16
issue
1
article number
5402
publisher
Nature Publishing Group
external identifiers
  • pmid:40571681
  • scopus:105009248886
ISSN
2041-1723
DOI
10.1038/s41467-025-61016-5
language
English
LU publication?
yes
id
83ffa650-882f-47c5-9f3d-ffbe1ec295cf
date added to LUP
2025-10-27 10:12:59
date last changed
2025-11-10 11:41:17
@article{83ffa650-882f-47c5-9f3d-ffbe1ec295cf,
  abstract     = {{<p>Frameshifts can be caused by specific combinations of tRNA and mRNA. The wildtype AGC-decoding E. coli tRNA<sup>Ser3</sup><sub>GCU</sub> has been shown to induce −1 ribosomal frameshifting on GCA alanine codons, and proposed to read a two-base codon instead of a canonical triplet. However, it has remained unclear whether this type of non-cognate decoding can be accommodated by the ribosome. Here, we perform single-particle cryo-EM reconstructions on E. coli 70S ribosomes with the frameshift-inducing tRNA<sup>Ser3</sup> bound to the non-cognate GCA codon or the cognate AGC codon in the ribosomal A site. The structures demonstrate that doublet decoding is made possible when A1493, the conserved monitoring base in 16S rRNA, mimics a first codon base, forming a Hoogsteen base pair with U36 from the anticodon and stacking with the mRNA. This interaction pushes the first two bases of the A-site codon in position for base pairing with C35 and G34 of the anticodon.</p>}},
  author       = {{Krishnaswamy, Shruthi and Akbar, Shirin and Larsson, Daniel S.D. and Chen, Yang and Selmer, Maria}},
  issn         = {{2041-1723}},
  language     = {{eng}},
  number       = {{1}},
  publisher    = {{Nature Publishing Group}},
  series       = {{Nature Communications}},
  title        = {{Doublet decoding of tRNA<sup>Ser3</sup> demonstrates plasticity of ribosomal decoding center}},
  url          = {{http://dx.doi.org/10.1038/s41467-025-61016-5}},
  doi          = {{10.1038/s41467-025-61016-5}},
  volume       = {{16}},
  year         = {{2025}},
}