Doublecortin engages the microtubule lattice through a cooperative binding mode involving its C-terminal domain

  1. Atefeh Rafiei
  2. Sofia Cruz Tetlalmatzi
  3. Claire H Edrington
  4. Linda Lee
  5. D Alex Crowder
  6. Daniel J Saltzberg
  7. Andrej Sali
  8. Gary Brouhard
  9. David C Schriemer  Is a corresponding author
  1. University of Calgary, Canada
  2. McGill University, Canada
  3. University of California, San Francisco, United States

Abstract

Doublecortin (DCX) is a microtubule (MT) associated protein that regulates MT structure and function during neuronal development and mutations in DCX lead to a spectrum of neurological disorders. The structural properties of MT-bound DCX that explain these disorders are incompletely determined. Here, we describe the molecular architecture of the DCX-MT complex through an integrative modeling approach that combines data from X-ray crystallography, cryo-EM and a high-fidelity chemical crosslinking method. We demonstrate that DCX interacts with MTs through its N-terminal domain and induces a lattice-dependent self-association involving the C-terminal structured domain and its disordered tail, in a conformation that favors an open, domain-swapped state. The networked state can accommodate multiple different attachment points on the MT lattice, all of which orient the C-terminal tails away from the lattice. As numerous disease mutations cluster in the C-terminus, and regulatory phosphorylations cluster in its-tail, our study shows that lattice-driven self-assembly is an important property of DCX.

Data availability

The DCX-MT integrative models, including final structures, modeling details, and input experimental data, were deposited into the PDB-dev repository for integrative models (www.pdb-dev.com) as follows: Dimeric DCX-MT (diagonal1): PDBDEV_00000071 Dimeric DCX-MT (lateral): PDBDEV_00000072 Dimeric DCX-MT (longitudinal): PDBDEV_00000073 Dimeric DCX-MT (diagonal2): PDBDEV_00000074 All LC-MS/MS data generated to support the findings of this study have been deposited with the ProteomeXchange Consortium with the dataset identifier PXD033167.

The following data sets were generated
The following previously published data sets were used

Article and author information

Author details

  1. Atefeh Rafiei

    Department of Chemistry, University of Calgary, Calgary, Canada
    Competing interests
    The authors declare that no competing interests exist.
  2. Sofia Cruz Tetlalmatzi

    Department of Biology, McGill University, Montreal, Canada
    Competing interests
    The authors declare that no competing interests exist.
  3. Claire H Edrington

    Department of Biology, McGill University, Montreal, Canada
    Competing interests
    The authors declare that no competing interests exist.
  4. Linda Lee

    Department of Biochemistry and Molecular Biology, University of Calgary, Calgary, Canada
    Competing interests
    The authors declare that no competing interests exist.
  5. D Alex Crowder

    Department of Biochemistry and Molecular Biology, University of Calgary, Calgary, Canada
    Competing interests
    The authors declare that no competing interests exist.
  6. Daniel J Saltzberg

    Department of Bioengineering and Therapeutic Sciences, University of California, San Francisco, San Francisco, United States
    Competing interests
    The authors declare that no competing interests exist.
  7. Andrej Sali

    Department of Bioengineering and Therapeutic Sciences, University of California, San Francisco, San Francisco, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-0435-6197
  8. Gary Brouhard

    Department of Biology, McGill University, Montreal, Canada
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-9101-1247
  9. David C Schriemer

    Department of Biochemistry and Molecular Biology, University of Calgary, Calgary, Canada
    For correspondence
    dschriem@ucalgary.ca
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-5202-1618

Funding

Canarie (RS-326)

  • David C Schriemer

Natural Sciences and Engineering Research Council of Canada (RGPIN-2017-04879)

  • David C Schriemer

Natural Sciences and Engineering Research Council of Canada (RGPIN-2020-04876)

  • Gary Brouhard

Canadian Institutes of Health Research (PJT-148702)

  • Gary Brouhard

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

Reviewing Editor

  1. Kassandra M Ori-McKenney, University of California, United States

Version history

  1. Received: January 28, 2021
  2. Preprint posted: February 18, 2021 (view preprint)
  3. Accepted: April 7, 2022
  4. Accepted Manuscript published: April 29, 2022 (version 1)
  5. Version of Record published: May 20, 2022 (version 2)

Copyright

© 2022, Rafiei 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. Atefeh Rafiei
  2. Sofia Cruz Tetlalmatzi
  3. Claire H Edrington
  4. Linda Lee
  5. D Alex Crowder
  6. Daniel J Saltzberg
  7. Andrej Sali
  8. Gary Brouhard
  9. David C Schriemer
(2022)
Doublecortin engages the microtubule lattice through a cooperative binding mode involving its C-terminal domain
eLife 11:e66975.
https://doi.org/10.7554/eLife.66975

Share this article

https://doi.org/10.7554/eLife.66975

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