1. Cell Biology

A gene-expression screen identifies a non-toxic sumoylation inhibitor that Mimics SUMO-less human LRH-1 in liver

  1. Miyuki Suzawa
  2. Diego A Miranda
  3. Karmela A Ramos
  4. Kenny K-H Ang
  5. Emily J Faivre
  6. Christopher G Wilson
  7. Laura Caboni
  8. Michelle R Arkin
  9. Yeong-Sang Kim
  10. Robert J Fletterick
  11. Aaron Diaz
  12. John S Schneekloth
  13. Holly A Ingraham  Is a corresponding author
  1. University of California, San Francisco, United States
  2. National Cancer Institute, United States
https://doi.org/10.7554/eLife.09003
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Cite this article
  1. Miyuki Suzawa
  2. Diego A Miranda
  3. Karmela A Ramos
  4. Kenny K-H Ang
  5. Emily J Faivre
  6. Christopher G Wilson
  7. Laura Caboni
  8. Michelle R Arkin
  9. Yeong-Sang Kim
  10. Robert J Fletterick
  11. Aaron Diaz
  12. John S Schneekloth
  13. Holly A Ingraham
(2015)
A gene-expression screen identifies a non-toxic sumoylation inhibitor that Mimics SUMO-less human LRH-1 in liver
eLife 4:e09003.
https://doi.org/10.7554/eLife.09003
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Abstract

SUMO-modification of nuclear proteins has profound effects on gene expression. However, non-toxic chemical tools that modulate sumoylation in cells are lacking. Here, to identify small molecule sumoylation inhibitors we developed a cell-based screen that focused on the well-sumoylated substrate, human Liver Receptor Homolog-1 (hLRH-1, NR5A2). Our primary gene-expression screen assayed two SUMO-sensitive transcripts, APOC3 and MUC1 that are upregulated by SUMO-less hLRH-1 or by siUBC9 knockdown, respectively. A polyphenol, tannic acid (TA) emerged as a potent sumoylation inhibitor in vitro (IC50 = 12.8 µM) and in cells. TA also increased hLRH-1 occupancy on SUMO-sensitive transcripts. Most significantly, when tested in humanized mouse primary hepatocytes, TA inhibits hLRH-1 sumoylation and induces SUMO-sensitive genes, thereby recapitulating the effects of expressing SUMO-less hLRH-1 in mouse liver. Our findings underscore the benefits of phenotypic screening for targeting post-translational modifications, and illustrate the potential utility of TA for probing the cellular consequences of sumoylation.

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Author details

  1. Miyuki Suzawa

    Department of Cellular and Molecular Pharmacology, University of California, San Francisco, San Francisco, United States
    Competing interests
    The authors declare that no competing interests exist.

  2. Diego A Miranda

    Department of Cellular and Molecular Pharmacology, University of California, San Francisco, San Francisco, United States
    Competing interests
    The authors declare that no competing interests exist.

  3. Karmela A Ramos

    Department of Cellular and Molecular Pharmacology, University of California, San Francisco, San Francisco, United States
    Competing interests
    The authors declare that no competing interests exist.

  4. Kenny K-H Ang

    Small Molecule Discovery Center, Department of Pharmaceutical Chemistry, University of California, San Francisco, San Francisco, United States
    Competing interests
    The authors declare that no competing interests exist.

  5. Emily J Faivre

    Department of Cellular and Molecular Pharmacology, University of California, San Francisco, San Francisco, United States
    Competing interests
    The authors declare that no competing interests exist.

  6. Christopher G Wilson

    Small Molecule Discovery Center, Department of Pharmaceutical Chemistry, University of California, San Francisco, San Francisco, United States
    Competing interests
    The authors declare that no competing interests exist.

  7. Laura Caboni

    Department of Biochemistry and Biophysics, University of California, San Francisco, San Francisco, United States
    Competing interests
    The authors declare that no competing interests exist.

  8. Michelle R Arkin

    Small Molecule Discovery Center, Department of Pharmaceutical Chemistry, University of California, San Francisco, San Francisco, United States
    Competing interests
    The authors declare that no competing interests exist.

  9. Yeong-Sang Kim

    Chemical Biology Laboratory, National Cancer Institute, Frederick, United States
    Competing interests
    The authors declare that no competing interests exist.

  10. Robert J Fletterick

    Department of Biochemistry and Biophysics, University of California, San Francisco, San Francisco, United States
    Competing interests
    The authors declare that no competing interests exist.

  11. Aaron Diaz

    Department of Epidemiology and Biostatistics, University of California, San Francisco, San Francisco, United States
    Competing interests
    The authors declare that no competing interests exist.

  12. John S Schneekloth

    Chemical Biology Laboratory, National Cancer Institute, Frederick, United States
    Competing interests
    The authors declare that no competing interests exist.

  13. Holly A Ingraham

    Department of Cellular and Molecular Pharmacology, University of California, San Francisco, San Francisco, United States
    For correspondence
    holly.ingraham@ucsf.edu
    Competing interests
    The authors declare that no competing interests exist.

Reviewing Editor

  1. Peter Tontonoz, Howard Hughes Medical Institute, University of California, Los Angeles, United States

Ethics

Animal experimentation: This study was performed in strict accordance with the recommendations in the Guide for the Care and Use of Laboratory Animals of the National Institutes of Health. All of the animals were handled according to approved institutional animal care and use committee (IACUC) protocols (#AN109147-01) of the University of California, San Francisco.Mice were euthanized in accordance with the UCSF Institutional Animal Care and Use Committee under Ingraham lab protocol.

Version history

  1. Received: May 26, 2015
  2. Accepted: October 20, 2015
  3. Accepted Manuscript published: December 11, 2015 (version 1)
  4. Version of Record published: January 27, 2016 (version 2)

Copyright

© 2015, Suzawa 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. Miyuki Suzawa
  2. Diego A Miranda
  3. Karmela A Ramos
  4. Kenny K-H Ang
  5. Emily J Faivre
  6. Christopher G Wilson
  7. Laura Caboni
  8. Michelle R Arkin
  9. Yeong-Sang Kim
  10. Robert J Fletterick
  11. Aaron Diaz
  12. John S Schneekloth
  13. Holly A Ingraham
(2015)
A gene-expression screen identifies a non-toxic sumoylation inhibitor that Mimics SUMO-less human LRH-1 in liver
eLife 4:e09003.
https://doi.org/10.7554/eLife.09003

Share this article

https://doi.org/10.7554/eLife.09003

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