Doublet decoding of tRNASer3 demonstrates plasticity of ribosomal decoding center

Abstract Frameshifts can be caused by specific combinations of tRNA and mRNA. The wildtype AGC-decoding E. coli tRNASer3 GCU 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 uncle...

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Main Authors: Shruthi Krishnaswamy, Shirin Akbar, Daniel S. D. Larsson, Yang Chen, Maria Selmer
Format: Article
Language:English
Published: Nature Portfolio 2025-06-01
Series:Nature Communications
Online Access:https://doi.org/10.1038/s41467-025-61016-5
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author Shruthi Krishnaswamy
Shirin Akbar
Daniel S. D. Larsson
Yang Chen
Maria Selmer
author_facet Shruthi Krishnaswamy
Shirin Akbar
Daniel S. D. Larsson
Yang Chen
Maria Selmer
author_sort Shruthi Krishnaswamy
collection DOAJ
description Abstract Frameshifts can be caused by specific combinations of tRNA and mRNA. The wildtype AGC-decoding E. coli tRNASer3 GCU 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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spelling doaj-art-02e3fd0e99644ac8bdaffd296cfae7b02025-08-20T03:27:10ZengNature PortfolioNature Communications2041-17232025-06-011611610.1038/s41467-025-61016-5Doublet decoding of tRNASer3 demonstrates plasticity of ribosomal decoding centerShruthi Krishnaswamy0Shirin Akbar1Daniel S. D. Larsson2Yang Chen3Maria Selmer4Department of Cell and Molecular Biology, Uppsala University, BMC, P.O. Box 596Department of Cell and Molecular Biology, Uppsala University, BMC, P.O. Box 596Department of Cell and Molecular Biology, Uppsala University, BMC, P.O. Box 596Department of Cell and Molecular Biology, Uppsala University, BMC, P.O. Box 596Department of Cell and Molecular Biology, Uppsala University, BMC, P.O. Box 596Abstract Frameshifts can be caused by specific combinations of tRNA and mRNA. The wildtype AGC-decoding E. coli tRNASer3 GCU 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.https://doi.org/10.1038/s41467-025-61016-5
spellingShingle Shruthi Krishnaswamy
Shirin Akbar
Daniel S. D. Larsson
Yang Chen
Maria Selmer
Doublet decoding of tRNASer3 demonstrates plasticity of ribosomal decoding center
Nature Communications
title Doublet decoding of tRNASer3 demonstrates plasticity of ribosomal decoding center
title_full Doublet decoding of tRNASer3 demonstrates plasticity of ribosomal decoding center
title_fullStr Doublet decoding of tRNASer3 demonstrates plasticity of ribosomal decoding center
title_full_unstemmed Doublet decoding of tRNASer3 demonstrates plasticity of ribosomal decoding center
title_short Doublet decoding of tRNASer3 demonstrates plasticity of ribosomal decoding center
title_sort doublet decoding of trnaser3 demonstrates plasticity of ribosomal decoding center
url https://doi.org/10.1038/s41467-025-61016-5
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