Article
- The EMBO Journal (2008) 27, 2580 - 2591
- doi:10.1038/emboj.2008.174
Published online: 4 September 2008
Subject Categories:
Cripto recruits Furin and PACE4 and controls Nodal trafficking during proteolytic maturation
Marie-Hélène Blanchet1,a, J Ann Le Good1,a, Daniel Mesnard1, Viola Oorschot2, Stéphane Baflast1, Gabriella Minchiotti3, Judith Klumperman2 and Daniel B Constam1
- Ecole Polytechnique Fédérale de Lausanne (EPFL), and Swiss Institute for Experimental Cancer Research (ISREC), Epalinges, Switzerland
- Department of Cell Biology, Cell Microscopy Center, Utrecht, The Netherlands
- Institute of Genetics and Biophysics 'A Buzzati-Traverso', Naples, Italy
Correspondence to:
Daniel B Constam, Ecole Polytechnique Fédérale de Lausanne (EPFL), Swiss Institute for Experimental Cancer Research (ISREC), 155, Chemin des Boveresses, S/Lausanne, 1066 Epalinges, Switzerland. Tel.: +41 21 692 5820; Fax: +41 21 652 6933; E-mail: daniel.constam@epfl.ch
aThese authors contributed equally to this work
Received 7 March 2008; Accepted 8 August 2008
Abstract
The glycosylphosphatidylinositol (GPI)-anchored proteoglycan Cripto binds Nodal and its type I receptor Alk4 to activate Smad2,3 transcription factors, but a role during Nodal precursor processing has not been described. We show that Cripto also binds the proprotein convertases Furin and PACE4 and localizes Nodal processing at the cell surface. When coexpressed as in early embryonic cells, Cripto and uncleaved Nodal already associated during secretion, and a Cripto-interacting region in the Nodal propeptide potentiated the effect of proteolytic maturation on Nodal signalling. Disruption of the trans-Golgi network (TGN) by brefeldin A blocked secretion, but export of Cripto and Nodal to the cell surface was not inhibited, indicating that Nodal is exposed to extracellular convertases before entering the TGN/endosomal system. Density fractionation and antibody uptake experiments showed that Cripto guides the Nodal precursor in detergent-resistant membranes to endocytic microdomains marked by GFP–Flotillin. We conclude that Nodal processing and endocytosis are coupled in signal-receiving cells.
Keywords:
- EGF-CFC,
- TGF
, - trafficking,
- unconventional exocytosis
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