1. Cell Biology
Download icon

Epidermal growth factor receptor neddylation is regulated by a desmosomal-COP9 (constitutive photomorphogenesis 9) signalosome complex

  1. Nicole Ann Najor
  2. Gillian Nicole Fitz
  3. Jennifer Leigh Koetsier
  4. Lisa Mary Godsel
  5. Lauren Veronica Albrecht
  6. Robert M Harmon
  7. Kathleen Janee Green  Is a corresponding author
  1. University of Detroit Mercy, United States
  2. Northwestern University, United States
Research Article
  • Cited 7
  • Views 1,893
  • Annotations
Cite this article as: eLife 2017;6:e22599 doi: 10.7554/eLife.22599

Abstract

Cell junctions are scaffolds that integrate mechanical and chemical signaling. We previously showed that a desmosomal cadherin promotes keratinocyte differentiation in an adhesion-independent manner by dampening Epidermal Growth Factor Receptor (EGFR) activity. Here we identify a potential mechanism by which desmosomes assist the de-neddylating COP9 signalosome (CSN) in attenuating EGFR through an association between the Cops3 subunit of the CSN and desmosomal components, Desmoglein1 (Dsg1) and Desmoplakin (Dp), to promote epidermal differentiation. Silencing CSN or desmosome components shifts the balance of EGFR modifications from ubiquitination to neddylation, inhibiting EGFR dynamics in response to an acute ligand stimulus. A reciprocal relationship between loss of Dsg1 and neddylated EGFR was observed in a carcinoma model, consistent with a role in sustaining EGFR activity during tumor progression. Identification of this previously unrecognized function of the CSN in regulating EGFR neddylation has broad-reaching implications for understanding how homeostasis is achieved in regenerating epithelia.

Article and author information

Author details

  1. Nicole Ann Najor

    Department of Biology, School of Engineering and Science, University of Detroit Mercy, Detroit, United States
    Competing interests
    The authors declare that no competing interests exist.
  2. Gillian Nicole Fitz

    Department of Pathology, Feinberg School of Medicine, Northwestern University, Chicago, United States
    Competing interests
    The authors declare that no competing interests exist.
  3. Jennifer Leigh Koetsier

    Department of Pathology, Feinberg School of Medicine, Northwestern University, Chicago, United States
    Competing interests
    The authors declare that no competing interests exist.
  4. Lisa Mary Godsel

    Department of Pathology, Feinberg School of Medicine, Northwestern University, Chicago, United States
    Competing interests
    The authors declare that no competing interests exist.
  5. Lauren Veronica Albrecht

    Department of Pathology, Feinberg School of Medicine, Northwestern University, Chicago, United States
    Competing interests
    The authors declare that no competing interests exist.
  6. Robert M Harmon

    Department of Pathology, Feinberg School of Medicine, Northwestern University, Chicago, United States
    Competing interests
    The authors declare that no competing interests exist.
  7. Kathleen Janee Green

    Department of Pathology, Feinberg School of Medicine, Northwestern University, Detroit, United States
    For correspondence
    kgreen@northwestern.edu
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-7332-5867

Funding

National Institute of Arthritis and Musculoskeletal and Skin Diseases (Postdoctoral Fellowship,F32AR066465)

  • Nicole Ann Najor

National Institute of General Medical Sciences (Graduate Student Training Grant,T32GM008061)

  • Robert M Harmon

American Heart Association (Predoctoral Fellowship)

  • Lauren Veronica Albrecht
  • Robert M Harmon

National Institute of Arthritis and Musculoskeletal and Skin Diseases (Research Program Grant,R01 AR041836)

  • Kathleen Janee Green

National Institute of Arthritis and Musculoskeletal and Skin Diseases (Research Program Grant,R37 AR043380)

  • Kathleen Janee Green

National Cancer Institute (Research Program Grant,R01 CA122151)

  • Kathleen Janee Green

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

Reviewing Editor

  1. Reinhard Fässler, Max Planck Institute of Biochemistry, Germany

Publication history

  1. Received: October 22, 2016
  2. Accepted: September 8, 2017
  3. Accepted Manuscript published: September 11, 2017 (version 1)
  4. Version of Record published: October 31, 2017 (version 2)

Copyright

© 2017, Najor 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

  • 1,893
    Page views
  • 247
    Downloads
  • 7
    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
    2. Microbiology and Infectious Disease
    Yong Fu et al.
    Research Article

    Toxoplasma gondii has evolved different developmental stages for disseminating during acute infection (i.e. tachyzoites) and for establishing chronic infection (i.e. bradyzoites). Calcium ion (Ca2+) signaling tightly regulates the lytic cycle of tachyzoites by controlling microneme secretion and motility to drive egress and cell invasion. However, the roles of Ca2+ signaling pathways in bradyzoites remain largely unexplored. Here we show that Ca2+ responses are highly restricted in bradyzoites and that they fail to egress in response to agonists. Development of dual-reporter parasites revealed dampened Ca2+ responses and minimal microneme secretion by bradyzoites induced in vitro or harvested from infected mice and tested ex vivo. Ratiometric Ca2+ imaging demonstrated lower Ca2+ basal levels, reduced magnitude, and slower Ca2+ kinetics in bradyzoites compared with tachyzoites stimulated with agonists. Diminished responses in bradyzoites were associated with down-regulation of Ca2+-ATPases involved in intracellular Ca2+ storage in the endoplasmic reticulum (ER) and acidocalcisomes. Once liberated from cysts by trypsin digestion, bradyzoites incubated in glucose plus Ca2+ rapidly restored their intracellular Ca2+ and ATP stores leading to enhanced gliding. Collectively, our findings indicate that intracellular bradyzoites exhibit dampened Ca2+ signaling and lower energy levels that restrict egress, and yet upon release they rapidly respond to changes in the environment to regain motility.

    1. Cell Biology
    Michelina Kierzek et al.
    Tools and Resources

    Fluorescent probes that change their spectral properties upon binding to small biomolecules, ions, or changes in the membrane potential (Vm) are invaluable tools to study cellular signaling pathways. Here, we introduce a novel technique for simultaneous recording of multiple probes at millisecond time resolution: frequency- and spectrally-tuned multiplexing (FASTM). Different from present multiplexing approaches, FASTM uses phase-sensitive signal detection, which renders various combinations of common probes for Vm and ions accessible for multiplexing. Using kinetic stopped-flow fluorimetry, we show that FASTM allows simultaneous recording of rapid changes in Ca2+, pH, Na+, and Vm with high sensitivity and minimal crosstalk. FASTM is also suited for multiplexing using single-cell microscopy and genetically-encoded FRET biosensors. Moreover, FASTM is compatible with opto-chemical tools to study signaling using light. Finally, we show that the exceptional time resolution of FASTM also allows resolving rapid chemical reactions. Altogether, FASTM opens new opportunities for interrogating cellular signaling.