Doublecortin and JIP3 are neural-specific counteracting regulators of dynein-mediated retrograde trafficking

  1. Lu Rao
  2. Peijun Li
  3. Xinglei Liu
  4. Qi Wang
  5. Alexander I Son
  6. Arne Gennerich  Is a corresponding author
  7. Judy Shih-Hwa Liu  Is a corresponding author
  8. Xiaoqin Fu  Is a corresponding author
  1. Albert Einstein College of Medicine, United States
  2. Second Affiliated Hospital & Yuying Children's Hospital of Wenzhou Medical University, China
  3. Brown University, United States
  4. Children's National Hospital, United States

Abstract

Mutations in the microtubule (MT)-binding protein doublecortin (DCX) or in the MT-based molecular motor dynein result in lissencephaly. However, a functional link between DCX and dynein has not been defined. Here, we demonstrate that DCX negatively regulates dynein-mediated retrograde transport in neurons from Dcx-/y or Dcx-/y;Dclk1-/- mice by reducing dynein's association with MTs and by disrupting the composition of the dynein motor complex. Previous work showed an increased binding of the adaptor protein C-Jun-amino-terminal kinase-interacting protein 3 (JIP3) to dynein in the absence of DCX. Using purified components, we demonstrate that JIP3 forms an active motor complex with dynein and its cofactor dynactin with two dyneins per complex. DCX competes with the binding of the second dynein, resulting in a velocity reduction of the complex. We conclude that DCX negatively regulates dynein-mediated retrograde transport through two critical interactions by regulating dynein binding to MTs and by regulating the composition of the dynein motor complex.

Data availability

All data generated or analysed during this study are included in the manuscript and supporting file; Source Data files have been provided for figure 2, 3, 4, figure 2-figure supplement 1 and table 1.

The following data sets were generated

Article and author information

Author details

  1. Lu Rao

    Department of Biochemistry, Albert Einstein College of Medicine, Bronx, United States
    Competing interests
    The authors declare that no competing interests exist.
  2. Peijun Li

    Second Affiliated Hospital & Yuying Children's Hospital of Wenzhou Medical University, Wenzhou, China
    Competing interests
    The authors declare that no competing interests exist.
  3. Xinglei Liu

    Department of Neurology, Brown University, Providence, United States
    Competing interests
    The authors declare that no competing interests exist.
  4. Qi Wang

    Second Affiliated Hospital & Yuying Children's Hospital of Wenzhou Medical University, Wenzhou, China
    Competing interests
    The authors declare that no competing interests exist.
  5. Alexander I Son

    Children's National Research Institute, Children's National Hospital, Washington, United States
    Competing interests
    The authors declare that no competing interests exist.
  6. Arne Gennerich

    Department of Biochemistry, Albert Einstein College of Medicine, Bronx, United States
    For correspondence
    arne.gennerich@einsteinmed.edu
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-8346-5473
  7. Judy Shih-Hwa Liu

    Department of Neurology, Brown University, Providence, United States
    For correspondence
    judy_liu@brown.edu
    Competing interests
    The authors declare that no competing interests exist.
  8. Xiaoqin Fu

    Second Affiliated Hospital & Yuying Children's Hospital of Wenzhou Medical University, Wenzhou, China
    For correspondence
    fuxq@wzhealth.com
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-6354-8960

Funding

National Natural Science Foundation of China (81971425)

  • Xiaoqin Fu

National Natural Science Foundation of China (81871035)

  • Peijun Li

Natural Science Foundation of Zhejiang Province (LZ09H090001)

  • Peijun Li

Natural Science Foundation of Zhejiang Province (LY20H040002)

  • Xiaoqin Fu

National Institutes of Health (R01GM098469)

  • Arne Gennerich

National Institutes of Health (R01NS114636)

  • Arne Gennerich

National Institutes of Health (RO1NS104428-01)

  • Judy Shih-Hwa Liu

Brain and Behavior Research Foundation

  • Judy Shih-Hwa Liu

Whitehall Foundation

  • Judy Shih-Hwa Liu

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

Ethics

Animal experimentation: All animal procedures were approved by the Committee on the Ethics of Animal Experiments of Wenzhou Medical University (Permit number: wydw2019-0723).

Copyright

© 2022, Rao 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

  • 1,380
    views
  • 228
    downloads
  • 13
    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. Lu Rao
  2. Peijun Li
  3. Xinglei Liu
  4. Qi Wang
  5. Alexander I Son
  6. Arne Gennerich
  7. Judy Shih-Hwa Liu
  8. Xiaoqin Fu
(2022)
Doublecortin and JIP3 are neural-specific counteracting regulators of dynein-mediated retrograde trafficking
eLife 11:e82218.
https://doi.org/10.7554/eLife.82218

Share this article

https://doi.org/10.7554/eLife.82218

Further reading

    1. Cell Biology
    2. Evolutionary Biology
    Paul Richard J Yulo, Nicolas Desprat ... Heather L Hendrickson
    Research Article

    Maintenance of rod-shape in bacterial cells depends on the actin-like protein MreB. Deletion of mreB from Pseudomonas fluorescens SBW25 results in viable spherical cells of variable volume and reduced fitness. Using a combination of time-resolved microscopy and biochemical assay of peptidoglycan synthesis, we show that reduced fitness is a consequence of perturbed cell size homeostasis that arises primarily from differential growth of daughter cells. A 1000-generation selection experiment resulted in rapid restoration of fitness with derived cells retaining spherical shape. Mutations in the peptidoglycan synthesis protein Pbp1A were identified as the main route for evolutionary rescue with genetic reconstructions demonstrating causality. Compensatory pbp1A mutations that targeted transpeptidase activity enhanced homogeneity of cell wall synthesis on lateral surfaces and restored cell size homeostasis. Mechanistic explanations require enhanced understanding of why deletion of mreB causes heterogeneity in cell wall synthesis. We conclude by presenting two testable hypotheses, one of which posits that heterogeneity stems from non-functional cell wall synthesis machinery, while the second posits that the machinery is functional, albeit stalled. Overall, our data provide support for the second hypothesis and draw attention to the importance of balance between transpeptidase and glycosyltransferase functions of peptidoglycan building enzymes for cell shape determination.

    1. Cell Biology
    2. Developmental Biology
    Pavan K Nayak, Arul Subramanian, Thomas F Schilling
    Research Article

    Mechanical forces play a critical role in tendon development and function, influencing cell behavior through mechanotransduction signaling pathways and subsequent extracellular matrix (ECM) remodeling. Here we investigate the molecular mechanisms by which tenocytes in developing zebrafish embryos respond to muscle contraction forces during the onset of swimming and cranial muscle activity. Using genome-wide bulk RNA sequencing of FAC-sorted tenocytes we identify novel tenocyte markers and genes involved in tendon mechanotransduction. Embryonic tendons show dramatic changes in expression of matrix remodeling associated 5b (mxra5b), matrilin1 (matn1), and the transcription factor kruppel-like factor 2a (klf2a), as muscles start to contract. Using embryos paralyzed either by loss of muscle contractility or neuromuscular stimulation we confirm that muscle contractile forces influence the spatial and temporal expression patterns of all three genes. Quantification of these gene expression changes across tenocytes at multiple tendon entheses and myotendinous junctions reveals that their responses depend on force intensity, duration and tissue stiffness. These force-dependent feedback mechanisms in tendons, particularly in the ECM, have important implications for improved treatments of tendon injuries and atrophy.