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MRNA stem-loops can pause the ribosome by hindering A-site tRNA binding.


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 the 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.

SUBMITTER: Bao C 

PROVIDER: S-EPMC7282821 | biostudies-literature | 2020 May

REPOSITORIES: biostudies-literature

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mRNA stem-loops can pause the ribosome by hindering A-site tRNA binding.

Bao Chen C   Loerch Sarah S   Ling Clarence C   Korostelev Andrei A AA   Grigorieff Nikolaus N   Ermolenko Dmitri N DN  

eLife 20200519


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 <i>E. coli dnaX</i> mRNA and the <i>gag-pol</i> transcript of Human Immunodeficiency Virus (HIV) perturb translation elo  ...[more]

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