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mRNA stem-loops can pause the ribosome by hindering A-site tRNA binding [preprint]

Bao, Chen
Loerch, Sarah
Ling, Clarence
Korostelev, Andrei A.
Grigorieff, Nikolaus
Ermolenko, Dmitri N.
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Abstract

Although the elongating ribosome is an efficient helicase, certain mRNA stem-loop structures are known to impede ribosome movement along mRNA and stimulate programmed ribosome frameshifting via mechanisms that are not well understood. Using biochemical and single-molecule Förster resonance energy transfer (smFRET) experiments, we studied how frameshift-inducing stem-loops from E. coli dnaX mRNA and the gag-pol transcript of Human Immunodeficiency Virus (HIV) perturb translation elongation. We find that upon encountering the ribosome, the stem-loops strongly inhibit A-site tRNA binding and ribosome intersubunit rotation that accompanies translation elongation. Electron cryo-microscopy (cryo-EM) reveals that the HIV stem-loop docks into the A site of the ribosome. Our results suggest that mRNA stem-loops can transiently escape ribosome helicase by binding to the A site. Thus, the stem-loops can modulate gene expression by sterically hindering tRNA binding and inhibiting translation elongation.

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bioRxiv 2020.02.05.936120; doi: https://doi.org/10.1101/2020.02.05.936120. Link to preprint on bioRxiv service.

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10.1101/2020.02.05.936120
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Now published in eLife doi: 10.7554/eLife.55799

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The copyright holder for this preprint (which was not peer-reviewed) is the author/funder. It is made available under a CC-BY-ND 4.0 International license.