Skip to main content
bioRxiv
  • Home
  • About
  • Submit
  • ALERTS / RSS
Advanced Search
New Results

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

Chen Bao, Sarah Loerch, Clarence Ling, Andrei A. Korostelev, Nikolaus Grigorieff, View ORCID ProfileDmitri N. Ermolenko
doi: https://doi.org/10.1101/2020.02.05.936120
Chen Bao
1Department of Biochemistry & Biophysics at School of Medicine and Dentistry and Center for RNA Biology, University of Rochester, Rochester, NY, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Sarah Loerch
2Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Clarence Ling
1Department of Biochemistry & Biophysics at School of Medicine and Dentistry and Center for RNA Biology, University of Rochester, Rochester, NY, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Andrei A. Korostelev
3RNA Therapeutics Institute, Department of Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School, 364 Plantation St., Worcester, MA 01605, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Nikolaus Grigorieff
2Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, VA, USA
3RNA Therapeutics Institute, Department of Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School, 364 Plantation St., Worcester, MA 01605, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Dmitri N. Ermolenko
1Department of Biochemistry & Biophysics at School of Medicine and Dentistry and Center for RNA Biology, University of Rochester, Rochester, NY, USA
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • ORCID record for Dmitri N. Ermolenko
  • For correspondence: dmitri_ermolenko@urmc.rochester.edu niko@grigorieff.org
  • Abstract
  • Full Text
  • Info/History
  • Metrics
  • Preview PDF
Loading

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.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-ND 4.0 International license.
Back to top
PreviousNext
Posted February 06, 2020.
Download PDF
Email

Thank you for your interest in spreading the word about bioRxiv.

NOTE: Your email address is requested solely to identify you as the sender of this article.

Enter multiple addresses on separate lines or separate them with commas.
mRNA stem-loops can pause the ribosome by hindering A-site tRNA binding
(Your Name) has forwarded a page to you from bioRxiv
(Your Name) thought you would like to see this page from the bioRxiv website.
CAPTCHA
This question is for testing whether or not you are a human visitor and to prevent automated spam submissions.
Share
mRNA stem-loops can pause the ribosome by hindering A-site tRNA binding
Chen Bao, Sarah Loerch, Clarence Ling, Andrei A. Korostelev, Nikolaus Grigorieff, Dmitri N. Ermolenko
bioRxiv 2020.02.05.936120; doi: https://doi.org/10.1101/2020.02.05.936120
Digg logo Reddit logo Twitter logo Facebook logo Google logo LinkedIn logo Mendeley logo
Citation Tools
mRNA stem-loops can pause the ribosome by hindering A-site tRNA binding
Chen Bao, Sarah Loerch, Clarence Ling, Andrei A. Korostelev, Nikolaus Grigorieff, Dmitri N. Ermolenko
bioRxiv 2020.02.05.936120; doi: https://doi.org/10.1101/2020.02.05.936120

Citation Manager Formats

  • BibTeX
  • Bookends
  • EasyBib
  • EndNote (tagged)
  • EndNote 8 (xml)
  • Medlars
  • Mendeley
  • Papers
  • RefWorks Tagged
  • Ref Manager
  • RIS
  • Zotero
  • Tweet Widget
  • Facebook Like
  • Google Plus One

Subject Area

  • Biochemistry
Subject Areas
All Articles
  • Animal Behavior and Cognition (3697)
  • Biochemistry (7801)
  • Bioengineering (5686)
  • Bioinformatics (21316)
  • Biophysics (10592)
  • Cancer Biology (8193)
  • Cell Biology (11954)
  • Clinical Trials (138)
  • Developmental Biology (6772)
  • Ecology (10411)
  • Epidemiology (2065)
  • Evolutionary Biology (13890)
  • Genetics (9719)
  • Genomics (13083)
  • Immunology (8158)
  • Microbiology (20037)
  • Molecular Biology (7865)
  • Neuroscience (43116)
  • Paleontology (321)
  • Pathology (1279)
  • Pharmacology and Toxicology (2264)
  • Physiology (3358)
  • Plant Biology (7242)
  • Scientific Communication and Education (1314)
  • Synthetic Biology (2009)
  • Systems Biology (5545)
  • Zoology (1130)