Regulation of mRNA translation by a photoriboswitch

  1. Kelly A Rotstan
  2. Michael M Abdelsayed
  3. Luiz F M Passalacqua
  4. Fabio Chizzolini
  5. Kasireddy Sudarshan
  6. A Richard Chamberlin
  7. Jiří Míšek  Is a corresponding author
  8. Andrej Luptak  Is a corresponding author
  1. University of California, Irvine, United States
  2. Charles University, Czech Republic

Abstract

Optogenetic tools have revolutionized the study of receptor-mediated processes, but such tools are lacking for RNA-controlled systems. In particular, light-activated regulatory RNAs are needed for spatiotemporal control of gene expression. To fill this gap, we used in vitro selection to isolate a novel riboswitch that selectively binds the trans isoform of a stiff-stilbene (amino-tSS)–a rapidly and reversibly photoisomerizing small molecule. Structural probing revealed that the RNA binds amino-tSS about 100-times stronger than the cis photoisoform (amino-cSS). In vitro and in vivo functional analysis showed that the riboswitch, termed Werewolf-1 (Were-1), inhibits translation of a downstream open reading frame when bound to amino-tSS. Photoisomerization of the ligand with a sub-millisecond pulse of light induced the protein expression. In contrast, amino-cSS supported protein expression, which was inhibited upon photoisomerization to amino-tSS. Reversible photoregulation of gene expression using a genetically encoded RNA will likely facilitate high-resolution spatiotemporal analysis of complex RNA processes.

Data availability

All data generated or analysed during this study are included in the manuscript and supporting files. Source data file has been provided for all figures in an Excel file.

Article and author information

Author details

  1. Kelly A Rotstan

    Department of Pharmaceutical Sciences, University of California, Irvine, Irvine, United States
    Competing interests
    The authors declare that no competing interests exist.
  2. Michael M Abdelsayed

    Department of Molecular Biology and Biochemistry, University of California, Irvine, Irvine, United States
    Competing interests
    The authors declare that no competing interests exist.
  3. Luiz F M Passalacqua

    Department of Pharmaceutical Sciences, University of California, Irvine, Irvine, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-5490-2427
  4. Fabio Chizzolini

    Department of Pharmaceutical Sciences, University of California, Irvine, Irvine, United States
    Competing interests
    The authors declare that no competing interests exist.
  5. Kasireddy Sudarshan

    Department of Organic Chemistry, Charles University, Prague, Czech Republic
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-7072-3226
  6. A Richard Chamberlin

    Department of Pharmaceutical Sciences, University of California, Irvine, Irvine, United States
    Competing interests
    The authors declare that no competing interests exist.
  7. Jiří Míšek

    Department of Organic Chemistry, Charles University, Prague, Czech Republic
    For correspondence
    jirimisek1@gmail.com
    Competing interests
    The authors declare that no competing interests exist.
  8. Andrej Luptak

    Department of Pharmaceutical Sciences, University of California, Irvine, Irvine, United States
    For correspondence
    aluptak@uci.edu
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-0632-5442

Funding

National Science Foundation (1804220)

  • Andrej Luptak

National Institutes of Health (R01GM094929)

  • Andrej Luptak

John Templeton Foundation

  • Andrej Luptak

Grantová Agentura České Republiky (17-25897Y)

  • Jiří Míšek

National Institutes of Health (Graduate Student Fellowship T32GM108561)

  • Kelly A Rotstan

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (Graduate Student Fellowship 99999.013571/2013-03)

  • Luiz F M Passalacqua

University of California, Irvine (Miguel Velez Scholarship)

  • Luiz F M Passalacqua

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Reviewing Editor

  1. Adrian Ferré-D'Amaré

Publication history

  1. Received: September 9, 2019
  2. Accepted: February 12, 2020
  3. Accepted Manuscript published: February 13, 2020 (version 1)
  4. Version of Record published: March 2, 2020 (version 2)
  5. Version of Record updated: March 11, 2020 (version 3)

Copyright

© 2020, Rotstan 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.

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  1. Kelly A Rotstan
  2. Michael M Abdelsayed
  3. Luiz F M Passalacqua
  4. Fabio Chizzolini
  5. Kasireddy Sudarshan
  6. A Richard Chamberlin
  7. Jiří Míšek
  8. Andrej Luptak
(2020)
Regulation of mRNA translation by a photoriboswitch
eLife 9:e51737.
https://doi.org/10.7554/eLife.51737
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