1. Biochemistry and Chemical Biology
  2. Cell Biology

Control of the structural landscape and neuronal proteotoxicity of mutant Huntingtin by domains flanking the polyQ tract

  1. Koning Shen
  2. Barbara Calamini
  3. Jonathan A Fauerbach
  4. Boxue Ma
  5. Sarah H Shahmoradian
  6. Ivana L Serrano Lachapel
  7. Wah Chiu
  8. Donald C Lo
  9. Judith Frydman  Is a corresponding author
  1. Stanford University, United States
  2. Sanofi R&D, France
  3. Miltenyi Biotec, Germany
  4. Baylor College of Medicine, United States
  5. Paul Scherrer Institute, Switzerland
  6. Duke University Medical Center, United States
https://doi.org/10.7554/eLife.18065
Share this article
Cite this article
  1. Koning Shen
  2. Barbara Calamini
  3. Jonathan A Fauerbach
  4. Boxue Ma
  5. Sarah H Shahmoradian
  6. Ivana L Serrano Lachapel
  7. Wah Chiu
  8. Donald C Lo
  9. Judith Frydman
(2016)
Control of the structural landscape and neuronal proteotoxicity of mutant Huntingtin by domains flanking the polyQ tract
eLife 5:e18065.
https://doi.org/10.7554/eLife.18065
  2. Download BibTeX
  3. Download .RIS

Abstract

Many neurodegenerative diseases are linked to amyloid aggregation. In Huntington's disease (HD), neurotoxicity correlates with increased aggregation propensity of a polyglutamine (polyQ) expansion in exon 1 of mutant huntingtin protein (mHtt). Here we establish how the domains flanking the polyQ tract shape the mHtt conformational landscape in vitro and in neurons. In vitro, the flanking domains have opposing effects on the conformation and stabilities of oligomers and amyloid fibrils. The N-terminal N17 promotes amyloid fibril formation, while the C-terminal Proline Rich Domain destabilizes fibrils and enhances oligomer formation. However, in neurons both domains act synergistically to engage protective chaperone and degradation pathways promoting mHtt proteostasis. Surprisingly, when proteotoxicity was assessed in rat corticostriatal brain slices, either flanking region alone sufficed to generate a neurotoxic conformation, while the polyQ tract alone exhibited minimal toxicity. Linking mHtt structural properties to its neuronal proteostasis should inform new strategies for neuroprotection in polyQ-expansion diseases.

Article and author information

Author details

  1. Koning Shen

    Department of Biology, Stanford University, Stanford, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-2607-449X

  2. Barbara Calamini

    Open Innovation Access Platform, Sanofi R&D, Strasbourg, France
    Competing interests
    The authors declare that no competing interests exist.

  3. Jonathan A Fauerbach

    Miltenyi Biotec, Cologne, Germany
    Competing interests
    The authors declare that no competing interests exist.

  4. Boxue Ma

    Verna and Marrs McLean Department of Biochemistry and Molecular Biology, Baylor College of Medicine, Houston, United States
    Competing interests
    The authors declare that no competing interests exist.

  5. Sarah H Shahmoradian

    Laboratory of Biomolecular Research, Paul Scherrer Institute, Villigen, Switzerland
    Competing interests
    The authors declare that no competing interests exist.

  6. Ivana L Serrano Lachapel

    Department of Biology, Stanford University, Stanford, United States
    Competing interests
    The authors declare that no competing interests exist.

  7. Wah Chiu

    Verna and Marrs McLean Department of Biochemistry and Molecular Biology, Baylor College of Medicine, Houston, United States
    Competing interests
    The authors declare that no competing interests exist.

  8. Donald C Lo

    Center for Drug Discovery, Department of Neurobiology, Duke University Medical Center, Durham, United States
    Competing interests
    The authors declare that no competing interests exist.

  9. Judith Frydman

    Department of Biology, Stanford University, Stanford, United States
    For correspondence
    jfrydman@stanford.edu
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-2302-6943

Funding

National Institute of General Medical Sciences (gm56433)

  • Koning Shen
  • Judith Frydman

National Institute of Neurological Disorders and Stroke (NS080514)

  • Barbara Calamini
  • Donald C Lo

NIH Office of the Director (pn2ey016525)

  • Sarah H Shahmoradian
  • Wah Chiu
  • Judith Frydman

National Institute of General Medical Sciences (gm103832)

  • Boxue Ma
  • Sarah H Shahmoradian
  • Wah Chiu

ellison medical foundation

  • Jonathan A Fauerbach
  • Judith Frydman

National Institute of Neurological Disorders and Stroke (NS092525)

  • Koning Shen
  • Judith Frydman

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

Reviewing Editor

  1. Jeffery W Kelly, The Scripps Research Institute, United States

Ethics

Animal experimentation: No human subjects. Animals were handled and killed in accordance with NIH guidelines and under approval and oversight of the Duke University institutional animal care and use committee (IACUC) to Don Lo. Protocol number A147-14-06.

Version history

  1. Received: May 21, 2016
  2. Accepted: October 17, 2016
  3. Accepted Manuscript published: October 18, 2016 (version 1)
  4. Version of Record published: December 2, 2016 (version 2)

Copyright

© 2016, Shen 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

  • 3,421
    views
  • 943
    downloads
  • 64
    citations

Views, downloads and citations are aggregated across all versions of this paper published by eLife.

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)

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

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

  1. Koning Shen
  2. Barbara Calamini
  3. Jonathan A Fauerbach
  4. Boxue Ma
  5. Sarah H Shahmoradian
  6. Ivana L Serrano Lachapel
  7. Wah Chiu
  8. Donald C Lo
  9. Judith Frydman
(2016)
Control of the structural landscape and neuronal proteotoxicity of mutant Huntingtin by domains flanking the polyQ tract
eLife 5:e18065.
https://doi.org/10.7554/eLife.18065

Share this article

https://doi.org/10.7554/eLife.18065

Categories and tags

Research organisms

Further reading

    1. Biochemistry and Chemical Biology
    2. Chromosomes and Gene Expression

    Human DDX6 regulates translation and decay of inefficiently translated mRNAs

    Ramona Weber, Chung-Te Chang
    Research Article

    Recent findings indicate that the translation elongation rate influences mRNA stability. One of the factors that has been implicated in this link between mRNA decay and translation speed is the yeast DEAD-box helicase Dhh1p. Here, we demonstrated that the human ortholog of Dhh1p, DDX6, triggers the deadenylation-dependent decay of inefficiently translated mRNAs in human cells. DDX6 interacts with the ribosome through the Phe-Asp-Phe (FDF) motif in its RecA2 domain. Furthermore, RecA2-mediated interactions and ATPase activity are both required for DDX6 to destabilize inefficiently translated mRNAs. Using ribosome profiling and RNA sequencing, we identified two classes of endogenous mRNAs that are regulated in a DDX6-dependent manner. The identified targets are either translationally regulated or regulated at the steady-state-level and either exhibit signatures of poor overall translation or of locally reduced ribosome translocation rates. Transferring the identified sequence stretches into a reporter mRNA caused translation- and DDX6-dependent degradation of the reporter mRNA. In summary, these results identify DDX6 as a crucial regulator of mRNA translation and decay triggered by slow ribosome movement and provide insights into the mechanism by which DDX6 destabilizes inefficiently translated mRNAs.

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

    Special Issue: Allosteric regulation of kinase activity

    Amy H Andreotti, Volker Dötsch
    Editorial

    The articles in this special issue highlight how modern cellular, biochemical, biophysical and computational techniques are allowing deeper and more detailed studies of allosteric kinase regulation.

    1. Biochemistry and Chemical Biology
    2. Microbiology and Infectious Disease

    Chemoproteomics validates selective targeting of Plasmodium M1 alanyl aminopeptidase as an antimalarial strategy

    Carlo Giannangelo, Matthew P Challis ... Darren J Creek
    Research Article

    New antimalarial drug candidates that act via novel mechanisms are urgently needed to combat malaria drug resistance. Here, we describe the multi-omic chemical validation of Plasmodium M1 alanyl metalloaminopeptidase as an attractive drug target using the selective inhibitor, MIPS2673. MIPS2673 demonstrated potent inhibition of recombinant Plasmodium falciparum (PfA-M1) and Plasmodium vivax (PvA-M1) M1 metalloaminopeptidases, with selectivity over other Plasmodium and human aminopeptidases, and displayed excellent in vitro antimalarial activity with no significant host cytotoxicity. Orthogonal label-free chemoproteomic methods based on thermal stability and limited proteolysis of whole parasite lysates revealed that MIPS2673 solely targets PfA-M1 in parasites, with limited proteolysis also enabling estimation of the binding site on PfA-M1 to within ~5 Å of that determined by X-ray crystallography. Finally, functional investigation by untargeted metabolomics demonstrated that MIPS2673 inhibits the key role of PfA-M1 in haemoglobin digestion. Combined, our unbiased multi-omic target deconvolution methods confirmed the on-target activity of MIPS2673, and validated selective inhibition of M1 alanyl metalloaminopeptidase as a promising antimalarial strategy.