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bioRxiv · 10.1101/2025.09.26.678779

Structural mechanism of mRNA decoding by mammalian GTPase GTPBP1

Susorov, D.·Miscicka, A.·Golovenko, D.·Loveland, A. B.·Zinoviev, A.·Pestova, T. V.·Korostelev, A. A.
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Abstract

GTP-binding protein 1 (GTPBP1) is a widespread translational GTPase closely related to elongation factor eEF1A. The loss of GTPBP1 leads to errors in neuronal development in animals and is associated with neurodegenerative disorders in humans. Although linked to translation and quality control mechanisms, GTPBP1 functions remain largely obscure. Similarly to eEF1A, GTPBP1 delivers cognate aminoacyl-tRNA to the ribosomal A site in a GTP-dependent manner, but GTP hydrolysis is not followed by rapid peptide bond formation, and GTPBP1-mediated elongation is slow. To establish the basis for GTPBP1 function, we determined cryo-EM structures of 80S ribosomal complexes bound to GTPBP1*aa-tRNA with GTP or the non-hydrolysable analog GDPCP. They revealed that the unique GTPBP1 architecture, including the additional eIF1/IF3-like N-terminal domain and the shoulder-interacting H-loop in place of the 2 helix of canonical GTPases, is responsible for establishing GTPBP1-specific interactions with tRNA and the ribosome, leading to slow GTPBP1 dissociation after GTP hydrolysis and thus delayed tRNA accommodation. Slow dissociation correlates with an extended proofreading stage resulting in higher accuracy of GTPBP1-mediated decoding and potentially allows GTPBP1 to elicit its putative quality control functions. GTPBP1 visualization provides the foundation for mapping and elucidating GTPBP1 mutations associated with human diseases.

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Susorov, D., Miscicka, A., Golovenko, D., Loveland, A. B., Zinoviev, A., Pestova, T. V., Korostelev, A. A.. 2025-09-26. Structural mechanism of mRNA decoding by mammalian GTPase GTPBP1. https://doi.org/10.1101/2025.09.26.678779

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