1. Biochemistry and Chemical Biology
  2. Structural Biology and Molecular Biophysics
Download icon

Modulating FOXO3 transcriptional activity by small, DBD-binding molecules

  1. Judith Hagenbuchner
  2. Veronika Obsilova
  3. Teresa Kaserer
  4. Nora Kaiser
  5. Bettina Rass
  6. Katarina Psenakova
  7. Vojtech Docekal
  8. Miroslava Alblova
  9. Klara Kohoutova
  10. Daniela Schuster
  11. Tatsiana Aneichyk
  12. Jan Vesely
  13. Petra Obexer
  14. Tomas Obsil  Is a corresponding author
  15. Michael J Ausserlechner  Is a corresponding author
  1. Medical University Innsbruck, Austria
  2. The Czech Academy of Sciences, Czech Republic
  3. University of Innsbruck, Austria
  4. Charles University, Czech Republic
  5. Charles University, Afghanistan
Research Article
  • Cited 3
  • Views 1,716
  • Annotations
Cite this article as: eLife 2019;8:e48876 doi: 10.7554/eLife.48876

Abstract

FOXO transcription factors are critical regulators of cell homeostasis and steer cell death, differentiation and longevity in mammalian cells. By combined pharmacophore-modelling-based in silico and fluorescence polarization-based screening we identified small molecules that physically interact with the DNA-binding domain (DBD) of FOXO3 and modulate the FOXO3 transcriptional program in human cells. The mode of interaction between compounds and the FOXO3-DBD was assessed via NMR spectroscopy and docking studies. We demonstrate that compounds S9 and its oxalate salt S9OX interfere with FOXO3 target promoter binding, gene transcription and modulate the physiologic program activated by FOXO3 in cancer cells. These small molecules prove the druggability of the FOXO-DBD and provide a structural basis for modulating these important homeostasis regulators in normal and malignant cells.

Article and author information

Author details

  1. Judith Hagenbuchner

    Department of Pediatrics II, Medical University Innsbruck, Innsbruck, Austria
    Competing interests
    The authors declare that no competing interests exist.
  2. Veronika Obsilova

    Department of Structural Biology of Signaling Proteins, The Czech Academy of Sciences, Prague, Czech Republic
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-4887-0323
  3. Teresa Kaserer

    Institute of Pharmacy, University of Innsbruck, Innsbruck, Austria
    Competing interests
    The authors declare that no competing interests exist.
  4. Nora Kaiser

    Department of Pediatrics I, Medical University Innsbruck, Innsbruck, Austria
    Competing interests
    The authors declare that no competing interests exist.
  5. Bettina Rass

    Department of Pediatrics I, Medical University Innsbruck, Innsbruck, Austria
    Competing interests
    The authors declare that no competing interests exist.
  6. Katarina Psenakova

    Department of Structural Biology of Signaling Proteins, The Czech Academy of Sciences, Prague, Czech Republic
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-8877-6599
  7. Vojtech Docekal

    Department of Organic Chemistry, Charles University, Prague, Czech Republic
    Competing interests
    The authors declare that no competing interests exist.
  8. Miroslava Alblova

    Department of Structural Biology of Signaling Proteins, The Czech Academy of Sciences, Prague, Czech Republic
    Competing interests
    The authors declare that no competing interests exist.
  9. Klara Kohoutova

    Department of Structural Biology of Signaling Proteins, The Czech Academy of Sciences, Prague, Czech Republic
    Competing interests
    The authors declare that no competing interests exist.
  10. Daniela Schuster

    Institute of Pharmacy, University of Innsbruck, Innsbruck, Austria
    Competing interests
    The authors declare that no competing interests exist.
  11. Tatsiana Aneichyk

    Division of Molecular Pathophysiology, Medical University Innsbruck, Innsbruck, Austria
    Competing interests
    The authors declare that no competing interests exist.
  12. Jan Vesely

    Department of Organic Chemistry, Charles University, Prague, Afghanistan
    Competing interests
    The authors declare that no competing interests exist.
  13. Petra Obexer

    Department of Pediatrics II, Medical University Innsbruck, Innsbruck, Austria
    Competing interests
    The authors declare that no competing interests exist.
  14. Tomas Obsil

    Department of Structural Biology of Signaling Proteins, The Czech Academy of Sciences, Prague, Czech Republic
    For correspondence
    obsil@natur.cuni.cz
    Competing interests
    The authors declare that no competing interests exist.
  15. Michael J Ausserlechner

    Department of Pediatrics I, Medical University Innsbruck, Innsbruck, Austria
    For correspondence
    michael.j.ausserlechner@i-med.ac.at
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-1015-2302

Funding

Austrian Science Fund (I3089-B28)

  • Judith Hagenbuchner
  • Veronika Obsilova
  • Tomas Obsil
  • Michael J Ausserlechner

Grantová Agentura České Republiky (17-33854L)

  • Veronika Obsilova
  • Tomas Obsil

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

Reviewing Editor

  1. Xavier Darzacq, University of California, Berkeley, United States

Publication history

  1. Received: May 29, 2019
  2. Accepted: December 1, 2019
  3. Accepted Manuscript published: December 2, 2019 (version 1)
  4. Accepted Manuscript updated: December 4, 2019 (version 2)
  5. Version of Record published: December 18, 2019 (version 3)
  6. Version of Record updated: January 15, 2020 (version 4)

Copyright

© 2019, Hagenbuchner 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.

Metrics

  • 1,716
    Page views
  • 301
    Downloads
  • 3
    Citations

Article citation count generated by polling the highest count across the following sources: Crossref, PubMed Central, Scopus.

Download links

A two-part list of links to download the article, or parts of the article, in various formats.

Downloads (link to download the article as PDF)

Download citations (links to download the citations from this article in formats compatible with various reference manager tools)

Open citations (links to open the citations from this article in various online reference manager services)

Further reading

    1. Biochemistry and Chemical Biology
    2. Neuroscience
    Ricardo M Santos, Anton Sirota
    Research Article Updated

    Cholinergic fast time-scale modulation of cortical physiology is critical for cognition, but direct local measurement of neuromodulators in vivo is challenging. Choline oxidase (ChOx)-based electrochemical biosensors have been used to capture fast cholinergic signals in behaving animals. However, these transients might be biased by local field potential and O2-evoked enzymatic responses. Using a novel Tetrode-based Amperometric ChOx (TACO) sensor, we performed highly sensitive and selective simultaneous measurement of ChOx activity (COA) and O2. In vitro and in vivo experiments, supported by mathematical modeling, revealed that non-steady-state enzyme responses to O2 give rise to phasic COA dynamics. This mechanism accounts for most of COA transients in the hippocampus, including those following locomotion bouts and sharp-wave/ripples. Our results suggest that it is unfeasible to probe phasic cholinergic signals under most behavioral paradigms with current ChOx biosensors. This confound is generalizable to any oxidase-based biosensor, entailing rigorous controls and new biosensor designs.

    1. Biochemistry and Chemical Biology
    2. Structural Biology and Molecular Biophysics
    Manoj K Rathinaswamy et al.
    Research Article

    Class I Phosphoinositide 3-kinases (PI3Ks) are master regulators of cellular functions, with the class IB PI3K catalytic subunit (p110g) playing key roles in immune signalling. p110g is a key factor in inflammatory diseases, and has been identified as a therapeutic target for cancers due to its immunomodulatory role. Using a combined biochemical/biophysical approach, we have revealed insight into regulation of kinase activity, specifically defining how immunodeficiency and oncogenic mutations of R1021 in the C-terminus can inactivate or activate enzyme activity. Screening of inhibitors using HDX-MS revealed that activation loop-binding inhibitors induce allosteric conformational changes that mimic those in the R1021C mutant. Structural analysis of advanced PI3K inhibitors in clinical development revealed novel binding pockets that can be exploited for further therapeutic development. Overall this work provides unique insights into regulatory mechanisms that control PI3Kg kinase activity, and shows a framework for the design of PI3K isoform and mutant selective inhibitors.