SNX9-induced membrane tubulation regulates CD28 cluster stability and signalling
Manuela Ecker,
Richard Schregle,
Natasha Kapoor-Kaushik,
Pascal Rossatti,
Verena M Betzler,
Daryan Kempe,
Maté Biro,
Nicholas Ariotti,
Gregory MI Redpath,
Jeremie Rossy
Affiliations
Manuela Ecker
EMBL Australia Node in Single Molecule Science, School of Medical Sciences and the ARC Centre of Excellence in Advanced Molecular Imaging, University of New South Wales, Sydney, Australia
Richard Schregle
Biotechnology Institute Thurgau (BITg) at the University of Konstanz, Kreuzlingen, Switzerland; Department of Biology, University of Konstanz, Konstanz, Germany
Natasha Kapoor-Kaushik
Electron Microscopy Unit, Mark Wainwright Analytical Centre, University of New South Wales, Sydney, Australia
Pascal Rossatti
Biotechnology Institute Thurgau (BITg) at the University of Konstanz, Kreuzlingen, Switzerland
Verena M Betzler
Biotechnology Institute Thurgau (BITg) at the University of Konstanz, Kreuzlingen, Switzerland
Daryan Kempe
EMBL Australia Node in Single Molecule Science, School of Medical Sciences and the ARC Centre of Excellence in Advanced Molecular Imaging, University of New South Wales, Sydney, Australia
EMBL Australia Node in Single Molecule Science, School of Medical Sciences and the ARC Centre of Excellence in Advanced Molecular Imaging, University of New South Wales, Sydney, Australia
Nicholas Ariotti
Electron Microscopy Unit, Mark Wainwright Analytical Centre, University of New South Wales, Sydney, Australia; Institute for Molecular Bioscience (IMB), University of Queensland, Brisbane, Australia
Gregory MI Redpath
EMBL Australia Node in Single Molecule Science, School of Medical Sciences and the ARC Centre of Excellence in Advanced Molecular Imaging, University of New South Wales, Sydney, Australia
EMBL Australia Node in Single Molecule Science, School of Medical Sciences and the ARC Centre of Excellence in Advanced Molecular Imaging, University of New South Wales, Sydney, Australia; Biotechnology Institute Thurgau (BITg) at the University of Konstanz, Kreuzlingen, Switzerland; Department of Biology, University of Konstanz, Konstanz, Germany
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.
