1. Cell Biology
  2. Genetics and Genomics
Download icon

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 School of Medicine, United States
  2. University of Utah, United States
  3. University of Pittsburgh, United States
  4. The Jackson Laboratory, United States
Research Article
  • Cited 8
  • Views 1,579
  • Annotations
Cite this article as: eLife 2020;9:e57831 doi: 10.7554/eLife.57831

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

    Department of Human Genetics, University of Utah School of Medicine, 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

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

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

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

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

    Department of 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

    Department of Human Genetics, University of Utah School of Medicine, 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

    Department of 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 School of Medicine, 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.

Reviewing Editor

  1. Hugo J Bellen, Baylor College of Medicine, United States

Publication history

  1. Received: April 14, 2020
  2. Accepted: December 12, 2020
  3. Accepted Manuscript published: December 14, 2020 (version 1)
  4. Accepted Manuscript updated: December 17, 2020 (version 2)
  5. Version of Record published: December 23, 2020 (version 3)

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

  • 1,579
    Page views
  • 156
    Downloads
  • 8
    Citations

Article citation count generated by polling the highest count across the following sources: Crossref, PubMed Central, Scopus.

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)

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

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

Further reading

    1. Cell Biology
    Natalya Pashkova et al.
    Research Article

    Attachment of ubiquitin (Ub) to cell surface proteins serves as a signal for internalization via clathrin-mediated endocytosis (CME). How ubiquitinated membrane proteins engage the internalization apparatus remains unclear. The internalization apparatus contains proteins such as Epsin and Eps15, which bind Ub, potentially acting as adaptors for Ub-based internalization signals. Here we show that additional components of the endocytic machinery including CALM, HIP1R, and Sla2 bind Ub via their N-terminal ANTH domain, a domain belonging to the superfamily of ENTH and VHS domains. Structural studies revealed that Ub binds with µM affinity to a unique C-terminal region within the ANTH domain not found in ENTH domains. Functional studies showed that combined loss of Ub-binding by ANTH-domain proteins and other Ub-binding domains within the yeast internalization apparatus caused defects in the Ub-dependent internalization of the GPCR Ste2 that was engineered to rely exclusively on Ub as an internalization signal. In contrast, these mutations had no effect on the internalization of Ste2 engineered to use an alternate Ub-independent internalization signal. These studies define new components of the internalization machinery that work collectively with Epsin and Eps15 to specify recognition of Ub as an internalization signal.

    1. Cell Biology
    Richa Sardana et al.
    Short Report

    Protein glycosylation in the Golgi is a sequential process that requires proper distribution of transmembrane glycosyltransferase enzymes in the appropriate Golgi compartments. Some of the cytosolic machinery required for the steady-state localization of some Golgi enzymes are known but existing models do not explain how many of these enzymes are localized. Here, we uncover the role of an integral membrane protein in yeast, Erd1, as a key facilitator of Golgi glycosyltransferase recycling by directly interacting with both the Golgi enzymes and the cytosolic receptor, Vps74. Loss of Erd1 function results in mislocalization of Golgi enzymes to the vacuole/lysosome. We present evidence that Erd1 forms an integral part of the recycling machinery and ensures productive recycling of several early Golgi enzymes. Our work provides new insights on how the localization of Golgi glycosyltransferases is spatially and temporally regulated, and is finely tuned to the cues of Golgi maturation.