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Syndecan–syntenin–ALIX regulates the biogenesis of exosomes

Nature Cell Biology volume 14, pages 677–685 (2012)Cite this article

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Abstract

The biogenesis of exosomes, small secreted vesicles involved in signalling processes, remains incompletely understood. Here, we report evidence that the syndecan heparan sulphate proteoglycans and their cytoplasmic adaptor syntenin control the formation of exosomes. Syntenin interacts directly with ALIX through LYPX(n)L motifs, similarly to retroviral proteins, and supports the intraluminal budding of endosomal membranes. Syntenin exosomes depend on the availability of heparan sulphate, syndecans, ALIX and ESCRTs, and impact on the trafficking and confinement of FGF signals. This study identifies a key role for syndecan–syntenin–ALIX in membrane transport and signalling processes.

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Figure 1: Syntenin interacts with ALIX, adapting ALIX to syndecan, and co-fractionates with these in exosomes.
Figure 2: Syntenin gain enhances exosome yields.
Figure 3: Syndecan–syntenin–ALIX depletion impairs exosome yields.
Figure 4: Syntenin exosomes are of endosomal origin.
Figure 5: The biogenesis of syntenin exosomes requires ESCRTs.
Figure 6: Syntenin exosomes require syndecan oligomerization and cleavage.

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Acknowledgements

We are grateful to H. Ceulemans, G. Reekmans and J. Grootjans for technical support, and to P. Baatsen for access to the electron microscopy facility at the Department for Human Genetics, VIB-KULeuven. We thank C. Dotti and P. Courtoy for critical feedback on the manuscript. This work was supported by the Fund for Scientific Research-Flanders (FWO), the Concerted Actions Program of the K.U. Leuven, the VIB, the Belgian Federation against Cancer (STK), the Interuniversity Attraction Poles of the Prime Ministers Services (IUAP) and the EMBO young investigator programme (P.Z.). A.M. was supported by a Marie Curie postdoctoral fellowship of the EU (FP7); Y.I. by an EMBO-long term postdoctoral fellowship; and A.G. by a PhD-student fellowship of the Agency for Innovation by Science and Technology (IWT), Flanders.

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Author notes

  1. Zhe Zhang

    Present address: Present address: State Key Laboratory of Molecular Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China,

  2. Maria Francesca Baietti, Zhe Zhang and Eva Mortier: These authors contributed equally to this work

Authors and Affiliations

  1. Department of Human Genetics, Laboratory for Glycobiology and Developmental Genetics, KULeuven, Campus Gasthuisberg, Herestraat 49, 3000 Leuven, Belgium

    Maria Francesca Baietti, Zhe Zhang, Aurélie Melchior, Gisèle Degeest, Christien Coomans & Guido David

  2. Department of Human Genetics, Laboratory for Signal Integration in Cell Fate Decision, KULeuven, Campus Gasthuisberg, Herestraat 49, 3000 Leuven, Belgium

    Maria Francesca Baietti, Eva Mortier, Annelies Geeraerts, Ylva Ivarsson, Fabienne Depoortere, Elke Vermeiren & Pascale Zimmermann

  3. Center for the Biology of Disease, VIB, 3000 Leuven, Belgium

    Maria Francesca Baietti, Zhe Zhang, Aurélie Melchior, Gisèle Degeest, Christien Coomans & Guido David

  4. Department of Chemistry, Laboratory for Biomolecular Dynamics, KULeuven, 3001 Heverlee, Belgium

    Annelies Geeraerts

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Contributions

The work is an equal contribution of two laboratories. M.F.B., Z.Z. and A.M. performed the cellular experiments; E.M. and Y.I. did the Biacore, A.G. the fluorescence spectroscopy, G. Degeest the electron microscopy, and F.D. the co-immunoprecipitation and the Y2H analyses; E.V. made the various expression constructs and C.C. assisted with cell culture and biochemical analyses; P.Z. and G. David designed experiments, analysed data and wrote the manuscript. All authors discussed results and commented on the manuscript.

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Correspondence to Pascale Zimmermann or Guido David.

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Baietti, M., Zhang, Z., Mortier, E. et al. Syndecan–syntenin–ALIX regulates the biogenesis of exosomes. Nat Cell Biol 14, 677–685 (2012). https://doi.org/10.1038/ncb2502

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