The ectodomains determine ligand function in vivo and selectivity of DLL1 and DLL4 toward NOTCH1 and NOTCH2 in vitro

  1. Lena Tveriakhina
  2. Karin Schuster-Gossler
  3. Sanchez M Jarrett
  4. Marie B Andrawes
  5. Meike Rohrbach
  6. Stephen C Blacklow  Is a corresponding author
  7. Achim Gossler  Is a corresponding author
  1. Medizinische Hochschule Hannover, Germany
  2. Harvard Medical School, United States

Abstract

DLL1 and DLL4 are Notch ligands with high structural similarity but context-dependent functional differences. Here, we analyze their functional divergence using cellular co-culture assays, biochemical studies, and in vivo experiments. DLL1 and DLL4 activate NOTCH1 and NOTCH2 differently in cell-based assays and this discriminating potential lies in the region between the N-terminus and EGF repeat three. Mice expressing chimeric ligands indicate that the ectodomains dictate ligand function during somitogenesis, and that during myogenesis even regions C-terminal to EGF3 are interchangeable. Substitution of NOTCH1-interface residues in the MNNL and DSL domains of DLL1 with the corresponding amino acids of DLL4, however, does not disrupt DLL1 function in vivo. Collectively, our data show that DLL4 preferentially activates NOTCH1 over NOTCH2, whereas DLL1 is equally effective in activating NOTCH1 and NOTCH2, establishing that the ectodomains dictate selective ligand function in vivo, and that features outside the known binding interface contribute to their differences.

Data availability

All data generated or analysed during this study are included in the manuscript and supporting files and source data files.

Article and author information

Author details

  1. Lena Tveriakhina

    Institut für Molekularbiologie, Medizinische Hochschule Hannover, Hannover, Germany
    Competing interests
    The authors declare that no competing interests exist.
  2. Karin Schuster-Gossler

    Institut für Molekularbiologie, Medizinische Hochschule Hannover, Hannover, Germany
    Competing interests
    The authors declare that no competing interests exist.
  3. Sanchez M Jarrett

    Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, United States
    Competing interests
    The authors declare that no competing interests exist.
  4. Marie B Andrawes

    Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, United States
    Competing interests
    The authors declare that no competing interests exist.
  5. Meike Rohrbach

    Institut für Molekularbiologie, Medizinische Hochschule Hannover, Hannover, Germany
    Competing interests
    The authors declare that no competing interests exist.
  6. Stephen C Blacklow

    Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, United States
    For correspondence
    stephen_blacklow@hms.harvard.edu
    Competing interests
    The authors declare that no competing interests exist.
  7. Achim Gossler

    Institut für Molekularbiologie, Medizinische Hochschule Hannover, Hannover, Germany
    For correspondence
    gossler.achim@mh-hannover.de
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-9103-9116

Funding

Deutsche Forschungsgemeinschaft (GO 449/13-1)

  • Achim Gossler

National Institutes of Health (R35-CA220340)

  • Stephen C Blacklow

van Maanen Graduate fellowship

  • Sanchez M Jarrett

Deutsche Forschungsgemeinschaft (REBIRTH)

  • Achim Gossler

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 experiments were performed according to the German rules and regulations (Tierschutzgesetz) and approved by the ethics committee of Lower Saxony for care and use of laboratory animals LAVES (Niedersächsisches Landesamt für Verbraucherschutz und Lebensmittelsicherheit; refs.: 33.12-42502-04-13/1314 and 33.14-42502-04-13/1293). Mice were housed in the central animal facility of Hannover Medical School (ZTL) and were maintained as approved by the responsible Veterinary Officer of the City of Hannover. Animal welfare was supervised and approved by the Institutional Animal Welfare Officer (Tierschutzbeauftragter).

Reviewing Editor

  1. Urban Lendahl, Karolinska Institute, Sweden

Publication history

  1. Received: July 13, 2018
  2. Accepted: October 1, 2018
  3. Accepted Manuscript published: October 5, 2018 (version 1)
  4. Version of Record published: October 25, 2018 (version 2)

Copyright

© 2018, Tveriakhina 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. Lena Tveriakhina
  2. Karin Schuster-Gossler
  3. Sanchez M Jarrett
  4. Marie B Andrawes
  5. Meike Rohrbach
  6. Stephen C Blacklow
  7. Achim Gossler
(2018)
The ectodomains determine ligand function in vivo and selectivity of DLL1 and DLL4 toward NOTCH1 and NOTCH2 in vitro
eLife 7:e40045.
https://doi.org/10.7554/eLife.40045
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