1. Cell Biology
  2. Genetics and Genomics

A Drosophila screen identifies NKCC1 as a modifier of NGLY1 deficiency

  1. Dana M Talsness  Is a corresponding author
  2. Katie G Owings
  3. Emily Coelho
  4. Gaelle Mercenne
  5. John M Pleinis
  6. Raghavendran Partha
  7. Kevin A Hope
  8. Aamir R Zuberi
  9. Nathan L Clark
  10. Cathleen M Lutz
  11. Aylin R Rodan
  12. Clement Y Chow  Is a corresponding author
  1. University of Utah, United States
  2. University of Pittsburgh, United States
  3. The Jackson Laboratory, United States
https://doi.org/10.7554/eLife.57831
Share this article
Cite this article
  1. Dana M Talsness
  2. Katie G Owings
  3. Emily Coelho
  4. Gaelle Mercenne
  5. John M Pleinis
  6. Raghavendran Partha
  7. Kevin A Hope
  8. Aamir R Zuberi
  9. Nathan L Clark
  10. Cathleen M Lutz
  11. Aylin R Rodan
  12. Clement Y Chow
(2020)
A Drosophila screen identifies NKCC1 as a modifier of NGLY1 deficiency
eLife 9:e57831.
https://doi.org/10.7554/eLife.57831
  2. Download BibTeX
  3. Download .RIS

Abstract

N-Glycanase 1 (NGLY1) is a cytoplasmic deglycosylating enzyme. Loss-of-function mutations in the NGLY1 gene cause NGLY1 deficiency, which is characterized by developmental delay, seizures, and a lack of sweat and tears. To model the phenotypic variability observed among patients, we crossed a Drosophila model of NGLY1 deficiency onto a panel of genetically diverse strains. The resulting progeny showed a phenotypic spectrum from 0-100% lethality. Association analysis on the lethality phenotype, as well as an evolutionary rate covariation analysis, generated lists of modifying genes, providing insight into NGLY1 function and disease. The top association hit was Ncc69 (human NKCC1/2), a conserved ion transporter. Analyses in NGLY1 -/- mouse cells demonstrated that NKCC1 has an altered average molecular weight and reduced function. The misregulation of this ion transporter may explain the observed defects in secretory epithelium function in NGLY1 deficiency patients.

Data availability

All data generated by this study are included in the manuscript and supporting files.

The following previously published data sets were used

Article and author information

Author details

  1. Dana M Talsness

    Human Genetics, University of Utah, Salt Lake City, United States
    For correspondence
    dana.talsness@genetics.utah.edu
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-7823-1616

  2. Katie G Owings

    Human Genetics, University of Utah, Salt Lake City, United States
    Competing interests
    The authors declare that no competing interests exist.

  3. Emily Coelho

    Human Genetics, University of Utah, Salt Lake City, United States
    Competing interests
    The authors declare that no competing interests exist.

  4. Gaelle Mercenne

    Internal Medicine, University of Utah, Salt Lake City, United States
    Competing interests
    The authors declare that no competing interests exist.

  5. John M Pleinis

    Internal Medicine, University of Utah, Salt Lake City, United States
    Competing interests
    The authors declare that no competing interests exist.

  6. Raghavendran Partha

    Computational and Systems Biology, University of Pittsburgh, Pittsburgh, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-7900-4375

  7. Kevin A Hope

    Human Genetics, University of Utah, Salt Lake City, United States
    Competing interests
    The authors declare that no competing interests exist.

  8. Aamir R Zuberi

    The Jackson Laboratory, Bar Harbor, United States
    Competing interests
    The authors declare that no competing interests exist.

  9. Nathan L Clark

    Department of Human Genetics, University of Utah, Salt Lake City, United States
    Competing interests
    The authors declare that no competing interests exist.

  10. Cathleen M Lutz

    The Jackson Laboratory, Bar Harbor, United States
    Competing interests
    The authors declare that no competing interests exist.

  11. Aylin R Rodan

    Internal Medicine, University of Utah, Salt Lake City, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-9202-2378

  12. Clement Y Chow

    Department of Human Genetics, University of Utah, Salt Lake City, United States
    For correspondence
    cchow@genetics.utah.edu
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-3104-7923

Funding

National Institute of General Medical Sciences (R35GM124780)

  • Clement Y Chow

National Institute of Diabetes and Digestive and Kidney Diseases (R01 DK110358)

  • Aylin R Rodan

National Human Genome Research Institute (R01 HG009299)

  • Nathan L Clark

Glenn Foundation for Medical Research (Glenn Award)

  • Clement Y Chow

National Human Genome Research Institute (T32 HG008962)

  • Dana M Talsness
  • Kevin A Hope

Might family (Bertrand T Might Fellowship)

  • Dana M Talsness

National Institute of General Medical Sciences (T32 GM007464)

  • Katie G Owings

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

Copyright

© 2020, Talsness 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

  • 2,620
    views
  • 256
    downloads
  • 32
    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. Dana M Talsness
  2. Katie G Owings
  3. Emily Coelho
  4. Gaelle Mercenne
  5. John M Pleinis
  6. Raghavendran Partha
  7. Kevin A Hope
  8. Aamir R Zuberi
  9. Nathan L Clark
  10. Cathleen M Lutz
  11. Aylin R Rodan
  12. Clement Y Chow
(2020)
A Drosophila screen identifies NKCC1 as a modifier of NGLY1 deficiency
eLife 9:e57831.
https://doi.org/10.7554/eLife.57831

Share this article

https://doi.org/10.7554/eLife.57831

Categories and tags

Research organisms

Further reading

    1. Cell Biology

    Ezrin defines TSC complex activation at endosomal compartments through EGFR–AKT signaling

    Giuliana Giamundo, Daniela Intartaglia ... Ivan Conte
    Research Article

    Endosomes have emerged as major signaling hubs where different internalized ligand–receptor complexes are integrated and the outcome of signaling pathways are organized to regulate the strength and specificity of signal transduction events. Ezrin, a major membrane–actin linker that assembles and coordinates macromolecular signaling complexes at membranes, has emerged recently as an important regulator of lysosomal function. Here, we report that endosomal-localized EGFR/Ezrin complex interacts with and triggers the inhibition of the Tuberous Sclerosis Complex (TSC complex) in response to EGF stimuli. This is regulated through activation of the AKT signaling pathway. Loss of Ezrin was not sufficient to repress TSC complex by EGF and culminated in translocation of TSC complex to lysosomes triggering suppression of mTORC1 signaling. Overexpression of constitutively active EZRINT567D is sufficient to relocalize TSC complex to the endosomes and reactivate mTORC1. Our findings identify EZRIN as a critical regulator of autophagy via TSC complex in response to EGF stimuli and establish the central role of early endosomal signaling in the regulation of mTORC1. Consistently, Medaka fish deficient for Ezrin exhibit defective endo-lysosomal pathway, attributable to the compromised EGFR/AKT signaling, ultimately leading to retinal degeneration. Our data identify a pivotal mechanism of endo-lysosomal signaling involving Ezrin and its associated EGFR/TSC complex, which are essential for retinal function.

    1. Cell Biology

    Beta Cells: Opportunity makes a hub or a leader

    Marjan Slak Rupnik
    Insight

    Functional subpopulations of β-cells emerge to control pulsative insulin secretion in the pancreatic islets of mice through calcium oscillations.

    1. Cell Biology

    Glucokinase activity controls peripherally located subpopulations of β-cells that lead islet Ca2+ oscillations

    Erli Jin, Jennifer K Briggs ... Matthew J Merrins
    Research Article

    Oscillations in insulin secretion, driven by islet Ca2+ waves, are crucial for glycemic control. Prior studies, performed with single-plane imaging, suggest that subpopulations of electrically coupled β-cells have privileged roles in leading and coordinating the propagation of Ca2+ waves. Here, we used three-dimensional (3D) light-sheet imaging to analyze the location and Ca2+ activity of single β-cells within the entire islet at >2 Hz. In contrast with single-plane studies, 3D network analysis indicates that the most highly synchronized β-cells are located at the islet center, and remain regionally but not cellularly stable between oscillations. This subpopulation, which includes ‘hub cells’, is insensitive to changes in fuel metabolism induced by glucokinase and pyruvate kinase activation. β-Cells that initiate the Ca2+ wave (leaders) are located at the islet periphery, and strikingly, change their identity over time via rotations in the wave axis. Glucokinase activation, which increased oscillation period, reinforced leader cells and stabilized the wave axis. Pyruvate kinase activation, despite increasing oscillation frequency, had no effect on leader cells, indicating the wave origin is patterned by fuel input. These findings emphasize the stochastic nature of the β-cell subpopulations that control Ca2+ oscillations and identify a role for glucokinase in spatially patterning ‘leader’ β-cells.