mRNA stem-loops can pause the ribosome by hindering A-site tRNA binding
Authors
Bao, ChenLoerch, Sarah
Ling, Clarence
Korostelev, Andrei A.
Grigorieff, Nikolaus
Ermolenko, Dmitri N.
UMass Chan Affiliations
Department of Biochemistry and Molecular PharmacologyRNA Therapeutics Institute
Document Type
Journal ArticlePublication Date
2020-05-19Keywords
E. colichromosomes
gene expression
molecular biophysics
structural biology
Biochemistry
Biophysics
Enzymes and Coenzymes
Molecular Biology
Nucleic Acids, Nucleotides, and Nucleosides
Structural Biology
Viruses
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Show full item recordAbstract
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 Forster 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.Source
Bao C, Loerch S, Ling C, Korostelev AA, Grigorieff N, Ermolenko DN. mRNA stem-loops can pause the ribosome by hindering A-site tRNA binding. Elife. 2020 May 19;9:e55799. doi: 10.7554/eLife.55799. Epub ahead of print. PMID: 32427100. Link to article on publisher's site
DOI
10.7554/eLife.55799Permanent Link to this Item
http://hdl.handle.net/20.500.14038/41462PubMed ID
32427100Related Resources
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© 2020, Bao et al. This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.Distribution License
http://creativecommons.org/licenses/by/4.0/ae974a485f413a2113503eed53cd6c53
10.7554/eLife.55799
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Except where otherwise noted, this item's license is described as © 2020, Bao et al. This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.