1. Biochemistry and Chemical Biology
  2. Cell Biology
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Endothelial PKA activity regulates angiogenesis by limiting autophagy through phosphorylation of ATG16L1

  1. Xiaocheng Zhao
  2. Pavel Nedvetsky
  3. Fabio Stanchi
  4. Anne-Clemence Vion
  5. Oliver Popp
  6. Kerstin Zühlke
  7. Gunnar Dittmar
  8. Enno Klussmann
  9. Holger Gerhardt  Is a corresponding author
  1. VIB, Belgium
  2. INSERM UMR-970, France
  3. Max Delbrück Center for Molecular Medicine, Germany
  4. Luxembourg Institute of Health, Luxembourg
Research Article
  • Cited 12
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Cite this article as: eLife 2019;8:e46380 doi: 10.7554/eLife.46380

Abstract

The cAMP-dependent protein kinase A (PKA) regulates various cellular functions in health and disease. In endothelial cells PKA activity promotes vessel maturation and limits tip cell formation. Here, we used a chemical genetic screen to identify endothelial-specific direct substrates of PKA in human umbilical vein endothelial cells (HUVEC) that may mediate these effects. Amongst several candidates, we identified ATG16L1, a regulator of autophagy, as novel target of PKA. Biochemical validation, mass spectrometry and peptide spot arrays revealed that PKA phosphorylates ATG16L1α at Ser268 and ATG16L1β at Ser269, driving phosphorylation-dependent degradation of ATG16L1 protein. Reducing PKA activity increased ATG16L1 protein levels and endothelial autophagy. Mouse in vivo genetics and pharmacological experiments demonstrated that autophagy inhibition partially rescues vascular hypersprouting caused by PKA deficiency. Together these results indicate that endothelial PKA activity mediates a critical switch from active sprouting to quiescence in part through phosphorylation of ATG16L1, which in turn reduces endothelial autophagy.

Data availability

The mass spectrometry proteomics data have been deposited to the ProteomeXchange Consortium via the PRIDE partner repository with the dataset identifier PXD012975. All data generated or analysed during this study are included in the manuscript and supporting files. Source data files have been provided for figures 3, 4 and 5.

The following data sets were generated

Article and author information

Author details

  1. Xiaocheng Zhao

    Vascular Patterning Laboratory, Center for Cancer Biology, VIB, Leuven, Belgium
    Competing interests
    No competing interests declared.

  2. Pavel Nedvetsky

    Vascular Patterning Laboratory, Center for Cancer Biology, VIB, Leuven, Belgium
    Competing interests
    No competing interests declared.

  3. Fabio Stanchi

    Vascular Patterning Laboratory, Center for Cancer Biology, VIB, Leuven, Belgium
    Competing interests
    No competing interests declared.

  4. Anne-Clemence Vion

    Paris Cardiovascular Research Center, INSERM UMR-970, Paris, France
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-2788-2512

  5. Oliver Popp

    Proteomics, Max Delbrück Center for Molecular Medicine, Berlin, Germany
    Competing interests
    No competing interests declared.

  6. Kerstin Zühlke

    Anchored Signaling Lab, Max Delbrück Center for Molecular Medicine, Berlin, Germany
    Competing interests
    No competing interests declared.

  7. Gunnar Dittmar

    Department of Oncology, Luxembourg Institute of Health, Luxembourg, Luxembourg
    Competing interests
    No competing interests declared.

  8. Enno Klussmann

    Anchored Signaling Lab, Max Delbrück Center for Molecular Medicine, Berlin, Germany
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-4004-5003

  9. Holger Gerhardt

    Integrative Vascular Biology Lab, Max Delbrück Center for Molecular Medicine, Berlin, Germany
    For correspondence
    holger.gerhardt@mdc-berlin.de
    Competing interests
    Holger Gerhardt, Reviewing editor, eLife.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-3030-0384

Funding

Fonds vor Wetenschappelijk Onderzoek (G.0742.15N)

  • Holger Gerhardt

Else-Kroener Stiftung (2014_A26)

  • Pavel Nedvetsky
  • Holger Gerhardt

European Research Council (311719 REshape)

  • Holger Gerhardt

Deutsche Forschungsgemeinschaft (DFG KL1415/7-1)

  • Enno Klussmann

Deutsche Forschungsgemeinschaft (394046635 - SFB 1365)

  • Enno Klussmann

Bundes Ministerium für Bildung und Forschung (16GW0179K)

  • Enno Klussmann

VIB (Tech Watch Q3 2015)

  • Pavel Nedvetsky
  • Holger Gerhardt

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 experimental procedures were approved by the Institutional Animal Care and Research Advisory Committee of the University of Leuven (application P249/2014) and performed according to the European guidelines.

Reviewing Editor

  1. Gou Young Koh, Institute of Basic Science and Korea Advanced Institute of Science and Technology (KAIST), Korea (South), Republic of

Publication history

  1. Received: February 25, 2019
  2. Accepted: October 1, 2019
  3. Accepted Manuscript published: October 3, 2019 (version 1)
  4. Version of Record published: October 17, 2019 (version 2)
  5. Version of Record updated: April 27, 2020 (version 3)

Copyright

© 2019, Zhao 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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