POMK regulates dystroglycan function via LARGE-mediated elongation of matriglycan

  1. Ameya S Walimbe
  2. Hidehiko Okuma
  3. Soumya Joseph
  4. Tiandi Yang
  5. Takahiro Yonekawa
  6. Jeffrey M Hord
  7. David Venzke
  8. Mary E Anderson
  9. Silvia Torelli
  10. Adnan Manzur
  11. Megan Devereaux
  12. Marco Cuellar
  13. Sally Prouty
  14. Saul Ocampo Landa
  15. Liping Yu
  16. Junyu Xiao
  17. Jack E Dixon
  18. Francesco Muntoni
  19. Kevin P Campbell  Is a corresponding author
  1. Howard Hughes Medical Institute, University of Iowa Roy J. and Lucille A. Carver College of Medicine, United States
  2. Dubowitz Neuromuscular Centre, UCL Great Ormond Street Institute of Child Health & Great Ormond Street Hospital, United Kingdom
  3. University of Iowa, United States
  4. Peking University, China
  5. University of California, San Diego, United States
  6. UCL Great Ormond Street Institute of Child Health, United Kingdom

Abstract

Matriglycan [-GlcA-β1,3-Xyl-α1,3-]n serves as a scaffold in many tissues for extracellular matrix proteins containing laminin-G domains including laminin, agrin, and perlecan. Like-acetylglucosaminyltransferase-1 (LARGE1) synthesizes and extends matriglycan on α-dystroglycan (α-DG) during skeletal muscle differentiation and regeneration; however, the mechanisms which regulate matriglycan elongation are unknown. Here, we show that Protein O-Mannose Kinase (POMK), which phosphorylates mannose of core M3 (GalNac-β1,3-GlcNac-β1,4-Man) preceding matriglycan synthesis, is required for LARGE1-mediated generation of full-length matriglycan on α-DG (~150 kDa). In the absence of Pomk in mouse skeletal muscle, LARGE1 synthesizes a very short matriglycan resulting in a ~90 kDa α-DG which binds laminin but cannot prevent eccentric contraction-induced force loss or muscle pathology. Solution NMR spectroscopy studies demonstrate that LARGE1 directly interacts with core M3 and binds preferentially to the phosphorylated form. Collectively, our study demonstrates that phosphorylation of core M3 by POMK enables LARGE1 to elongate matriglycan on α-DG, thereby preventing muscular dystrophy.

Data availability

All data generated or analysed during this study are included in the manuscript.

Article and author information

Author details

  1. Ameya S Walimbe

    Department of Molecular Physiology and Biophysics, Howard Hughes Medical Institute, University of Iowa Roy J. and Lucille A. Carver College of Medicine, Iowa City, United States
    Competing interests
    The authors declare that no competing interests exist.
  2. Hidehiko Okuma

    Department of Molecular Physiology and Biophysics, Howard Hughes Medical Institute, University of Iowa Roy J. and Lucille A. Carver College of Medicine, Iowa City, United States
    Competing interests
    The authors declare that no competing interests exist.
  3. Soumya Joseph

    Department of Molecular Physiology and Biophysics, Howard Hughes Medical Institute, University of Iowa Roy J. and Lucille A. Carver College of Medicine, Iowa City, United States
    Competing interests
    The authors declare that no competing interests exist.
  4. Tiandi Yang

    Department of Molecular Physiology and Biophysics, Howard Hughes Medical Institute, University of Iowa Roy J. and Lucille A. Carver College of Medicine, Iowa City, United States
    Competing interests
    The authors declare that no competing interests exist.
  5. Takahiro Yonekawa

    Department of Molecular Physiology and Biophysics, Howard Hughes Medical Institute, University of Iowa Roy J. and Lucille A. Carver College of Medicine, Iowa City, United States
    Competing interests
    The authors declare that no competing interests exist.
  6. Jeffrey M Hord

    Department of Molecular Physiology and Biophysics, Howard Hughes Medical Institute, University of Iowa Roy J. and Lucille A. Carver College of Medicine, Iowa City, United States
    Competing interests
    The authors declare that no competing interests exist.
  7. David Venzke

    Department of Molecular Physiology and Biophysics, Howard Hughes Medical Institute, University of Iowa Roy J. and Lucille A. Carver College of Medicine, Iowa City, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-8180-9562
  8. Mary E Anderson

    Department of Molecular Physiology and Biophysics, Howard Hughes Medical Institute, University of Iowa Roy J. and Lucille A. Carver College of Medicine, Iowa City, United States
    Competing interests
    The authors declare that no competing interests exist.
  9. Silvia Torelli

    Neurology, Dubowitz Neuromuscular Centre, UCL Great Ormond Street Institute of Child Health & Great Ormond Street Hospital, London, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
  10. Adnan Manzur

    Neurology, Dubowitz Neuromuscular Centre, UCL Great Ormond Street Institute of Child Health & Great Ormond Street Hospital, London, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
  11. Megan Devereaux

    Department of Molecular Physiology and Biophysics, Howard Hughes Medical Institute, University of Iowa Roy J. and Lucille A. Carver College of Medicine, Iowa City, United States
    Competing interests
    The authors declare that no competing interests exist.
  12. Marco Cuellar

    Department of Molecular Physiology and Biophysics, Howard Hughes Medical Institute, University of Iowa Roy J. and Lucille A. Carver College of Medicine, Iowa City, United States
    Competing interests
    The authors declare that no competing interests exist.
  13. Sally Prouty

    Department of Molecular Physiology and Biophysics, Howard Hughes Medical Institute, University of Iowa Roy J. and Lucille A. Carver College of Medicine, Iowa City, United States
    Competing interests
    The authors declare that no competing interests exist.
  14. Saul Ocampo Landa

    Department of Molecular Physiology and Biophysics, Howard Hughes Medical Institute, University of Iowa Roy J. and Lucille A. Carver College of Medicine, Iowa City, United States
    Competing interests
    The authors declare that no competing interests exist.
  15. Liping Yu

    University of Iowa Roy J. and Lucille A. Carver College of Medicine, University of Iowa, Iowa City, United States
    Competing interests
    The authors declare that no competing interests exist.
  16. Junyu Xiao

    School of Life Sciences, Peking University, Beijing, China
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-1822-1701
  17. Jack E Dixon

    Department of Pharmacology, University of California, San Diego, La Jolla, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-8266-5449
  18. Francesco Muntoni

    UCL Great Ormond Street Institute of Child Health, London, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
  19. Kevin P Campbell

    Department of Molecular Physiology and Biophysics, Howard Hughes Medical Institute, University of Iowa Roy J. and Lucille A. Carver College of Medicine, Iowa City, United States
    For correspondence
    kevin-campbell@uiowa.edu
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-2066-5889

Funding

Paul D. Wellstone Muscular Dystrophy Specialized Research Center grant (1U54NS053672)

  • Kevin P Campbell

Great Ormond Street Hospital for Children NHS Foundation Trust and University College

  • Silvia Torelli
  • Francesco Muntoni

Medical Scientist Training Program Grant by the National Institute of General Medical Sciences (5 T32 GM007337)

  • Ameya S Walimbe

Howard Hughes Medical Institute

  • Kevin P Campbell

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

Ethics

Animal experimentation: 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 animal experiments were approved by the Institutional Animal Care and Use Committee (IACUC) protocols of the University of Iowa (#0081122).

Human subjects: All procedures performed in this study involving human participants were in accordance with the ethical standards of NHS Health Research Authority (REC ref: 06/Q0406/33). We acknowledge and thank the BRC/MRC Centre for Neuromuscular Diseases Biobank for providing patients' serum and biopsy samples. We confirm that informed consent was provided to the patient and family regarding the nature of the genetic studies to be performed upon collection of samples and is available for our patient.

Reviewing Editor

  1. Joseph G Gleeson, Howard Hughes Medical Institute, The Rockefeller University, United States

Publication history

  1. Received: July 23, 2020
  2. Accepted: September 24, 2020
  3. Accepted Manuscript published: September 25, 2020 (version 1)
  4. Accepted Manuscript updated: September 29, 2020 (version 2)
  5. Version of Record published: October 14, 2020 (version 3)
  6. Version of Record updated: October 22, 2020 (version 4)

Copyright

© 2020, Walimbe 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. Ameya S Walimbe
  2. Hidehiko Okuma
  3. Soumya Joseph
  4. Tiandi Yang
  5. Takahiro Yonekawa
  6. Jeffrey M Hord
  7. David Venzke
  8. Mary E Anderson
  9. Silvia Torelli
  10. Adnan Manzur
  11. Megan Devereaux
  12. Marco Cuellar
  13. Sally Prouty
  14. Saul Ocampo Landa
  15. Liping Yu
  16. Junyu Xiao
  17. Jack E Dixon
  18. Francesco Muntoni
  19. Kevin P Campbell
(2020)
POMK regulates dystroglycan function via LARGE-mediated elongation of matriglycan
eLife 9:e61388.
https://doi.org/10.7554/eLife.61388
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