SNX9-induced membrane tubulation regulates CD28 cluster stability and signalling

  1. Ecker Ecker
  2. Richard Schregle
  3. Natasha Kapoor-Kaushik
  4. Pascal Rossatti
  5. Verena M Betzler
  6. Daryan Kempe
  7. Maté Biro
  8. Nicholas Ariotti
  9. Gregory MI Redpath  Is a corresponding author
  10. Jérémie Rossy  Is a corresponding author
  1. University of New South Wales, Australia
  2. University of Konstanz, Switzerland
  3. The University of New South Wales, Australia

Abstract

T cell activation requires engagement of a cognate antigen by the T cell receptor (TCR) and the co-stimulatory signal of CD28. Both TCR and CD28 aggregate into clusters at the plasma membrane of activated T cells. While the role of TCR clustering in T cell activation has been extensively investigated, little is known about how CD28 clustering contributes to CD28 signalling. Here we report that upon CD28 triggering, the BAR-domain protein sorting nexin 9 (SNX9) is recruited to CD28 clusters at the immunological synapse. Using three-dimensional correlative light and electron microscopy, we show that SNX9 generates membrane tubulation out of CD28 clusters. Our data further reveal that CD28 clusters are in fact dynamic structures and that SNX9 regulates their stability as well as CD28 phosphorylation and the resulting production of the cytokine IL-2. In summary, our work suggests a model in which SNX9-mediated tubulation generates a membrane environment that promotes CD28 triggering and downstream signalling events.

Data availability

All datasets for this study are deposited on Zenodo and are publicly available under a Creative Commons Attribution 4.0 International license

The following data sets were generated

Article and author information

Author details

  1. Ecker Ecker

    EMBL Australia Node in Single Molecule Science, University of New South Wales, Sydney, Australia
    Competing interests
    The authors declare that no competing interests exist.
  2. Richard Schregle

    Biotechnology Institute Thurgau, University of Konstanz, Kreuzlingen, Switzerland
    Competing interests
    The authors declare that no competing interests exist.
  3. Natasha Kapoor-Kaushik

    Electron Microscope Unit, University of New South Wales, Sydney, Australia
    Competing interests
    The authors declare that no competing interests exist.
  4. Pascal Rossatti

    Biotechnology Institute Thurgau, University of Konstanz, Kreuzlingen, Switzerland
    Competing interests
    The authors declare that no competing interests exist.
  5. Verena M Betzler

    Biotechnology Institute Thurgau, University of Konstanz, Kreuzlingen, Switzerland
    Competing interests
    The authors declare that no competing interests exist.
  6. Daryan Kempe

    EMBL Australia Node in Single Molecule Science, University of New South Wales, Sydney, Australia
    Competing interests
    The authors declare that no competing interests exist.
  7. Maté Biro

    EMBL Australia Node in Single Molecule Science, University of New South Wales, Sydney, Australia
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-5852-3726
  8. Nicholas Ariotti

    Electron Microscope Unit, University of New South Wales, Sydney, Australia
    Competing interests
    The authors declare that no competing interests exist.
  9. Gregory MI Redpath

    EMBL Australia Node in Single Molecule Science, The University of New South Wales, Sydney, Australia
    For correspondence
    gregmi.redpath@gmail.com
    Competing interests
    The authors declare that no competing interests exist.
  10. Jérémie Rossy

    Biotechnology Institute Thurgau, University of Konstanz, Kreuzlingen, Switzerland
    For correspondence
    jeremie.rossy@bitg.ch
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-5128-5283

Funding

Schweizerischer Nationalfonds zur Förderung der Wissenschaftlichen Forschung (31003A_172969)

  • Jérémie Rossy

Deutsche Forschungsgemeinschaft (RO 6238/1-1)

  • Jérémie Rossy

National Health and Medical Research Council (APP1102730)

  • Jérémie Rossy

Novartis Stiftung für Medizinisch-Biologische Forschung

  • Jérémie Rossy

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

Reviewing Editor

  1. Bernard Malissen, Centre d'Immunologie de Marseille-Luminy, Aix Marseille Université, France

Version history

  1. Received: February 15, 2021
  2. Accepted: January 12, 2022
  3. Accepted Manuscript published: January 20, 2022 (version 1)
  4. Version of Record published: January 21, 2022 (version 2)

Copyright

© 2022, Ecker 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

  • 1,683
    views
  • 203
    downloads
  • 5
    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. Ecker Ecker
  2. Richard Schregle
  3. Natasha Kapoor-Kaushik
  4. Pascal Rossatti
  5. Verena M Betzler
  6. Daryan Kempe
  7. Maté Biro
  8. Nicholas Ariotti
  9. Gregory MI Redpath
  10. Jérémie Rossy
(2022)
SNX9-induced membrane tubulation regulates CD28 cluster stability and signalling
eLife 11:e67550.
https://doi.org/10.7554/eLife.67550

Share this article

https://doi.org/10.7554/eLife.67550

Further reading

    1. Cell Biology
    2. Chromosomes and Gene Expression
    Lucie Crhak Khaitova, Pavlina Mikulkova ... Karel Riha
    Research Article

    Heat stress is a major threat to global crop production, and understanding its impact on plant fertility is crucial for developing climate-resilient crops. Despite the known negative effects of heat stress on plant reproduction, the underlying molecular mechanisms remain poorly understood. Here, we investigated the impact of elevated temperature on centromere structure and chromosome segregation during meiosis in Arabidopsis thaliana. Consistent with previous studies, heat stress leads to a decline in fertility and micronuclei formation in pollen mother cells. Our results reveal that elevated temperature causes a decrease in the amount of centromeric histone and the kinetochore protein BMF1 at meiotic centromeres with increasing temperature. Furthermore, we show that heat stress increases the duration of meiotic divisions and prolongs the activity of the spindle assembly checkpoint during meiosis I, indicating an impaired efficiency of the kinetochore attachments to spindle microtubules. Our analysis of mutants with reduced levels of centromeric histone suggests that weakened centromeres sensitize plants to elevated temperature, resulting in meiotic defects and reduced fertility even at moderate temperatures. These results indicate that the structure and functionality of meiotic centromeres in Arabidopsis are highly sensitive to heat stress, and suggest that centromeres and kinetochores may represent a critical bottleneck in plant adaptation to increasing temperatures.

    1. Cell Biology
    Wan-ping Yang, Mei-qi Li ... Qian-qian Luo
    Research Article

    High-altitude polycythemia (HAPC) affects individuals living at high altitudes, characterized by increased red blood cells (RBCs) production in response to hypoxic conditions. The exact mechanisms behind HAPC are not fully understood. We utilized a mouse model exposed to hypobaric hypoxia (HH), replicating the environmental conditions experienced at 6000 m above sea level, coupled with in vitro analysis of primary splenic macrophages under 1% O2 to investigate these mechanisms. Our findings indicate that HH significantly boosts erythropoiesis, leading to erythrocytosis and splenic changes, including initial contraction to splenomegaly over 14 days. A notable decrease in red pulp macrophages (RPMs) in the spleen, essential for RBCs processing, was observed, correlating with increased iron release and signs of ferroptosis. Prolonged exposure to hypoxia further exacerbated these effects, mirrored in human peripheral blood mononuclear cells. Single-cell sequencing showed a marked reduction in macrophage populations, affecting the spleen’s ability to clear RBCs and contributing to splenomegaly. Our findings suggest splenic ferroptosis contributes to decreased RPMs, affecting erythrophagocytosis and potentially fostering continuous RBCs production in HAPC. These insights could guide the development of targeted therapies for HAPC, emphasizing the importance of splenic macrophages in disease pathology.