title: Cripto recruits Furin and PACE4 and controls Nodal trafficking during proteolytic maturation.
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.
authors.affiliation: Ecole Polytechnique Fédérale de Lausanne (EPFL), Swiss Institute for Experimental Cancer Research (ISREC), Epalinges, Switzerland.
authors.forename: Marie-Hélène
authors.initials: MH
authors.lastname: Blanchet
authors.forename: J Ann
authors.initials: JA
authors.lastname: Le Good
authors.forename: Daniel
authors.initials: D
authors.lastname: Mesnard
authors.forename: Viola
authors.initials: V
authors.lastname: Oorschot
authors.forename: Stéphane
authors.initials: S
authors.lastname: Baflast
authors.forename: Gabriella
authors.initials: G
authors.lastname: Minchiotti
authors.forename: Judith
authors.initials: J
authors.lastname: Klumperman
authors.forename: Daniel B
authors.initials: DB
authors.lastname: Constam
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D019556:COS Cells; D002522:Chlorocebus aethiops; D011992:Endosomes / Q000378:metabolism; D004815:Epidermal Growth Factor / Q000235:genetics / Q000378:metabolism*; D005089:Exocytosis / Q000502:physiology; D045683:Furin / Q000235:genetics / Q000378:metabolism*; D017261:Glycosylphosphatidylinositols / Q000378:metabolism; D006801:Humans; D008562:Membrane Glycoproteins / Q000235:genetics / Q000378:metabolism*; D021962:Membrane Microdomains / Q000378:metabolism; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism; D051379:Mice; D008969:Molecular Sequence Data; D009363:Neoplasm Proteins / Q000235:genetics / Q000378:metabolism*; D055457:Nodal Protein; D043484:Proprotein Convertases / Q000235:genetics / Q000378:metabolism*; D011498:Protein Precursors / Q000378:metabolism; D021381:Protein Transport / Q000502:physiology; D016415:Sequence Alignment; D012697:Serine Endopeptidases / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction / Q000502:physiology; D016212:Transforming Growth Factor beta / Q000235:genetics / Q000378:metabolism*; D021601:trans-Golgi Network / Q000378:metabolism
chemical_list: D017261:Glycosylphosphatidylinositols; D008562:Membrane Glycoproteins; D008565:Membrane Proteins; C526007:NODAL protein, human; D009363:Neoplasm Proteins; D055457:Nodal Protein; C526008:Nodal protein, mouse; D011498:Protein Precursors; C489450:Tdgf1 protein, mouse; D016212:Transforming Growth Factor beta; C106354:flotillins; D004815:Epidermal Growth Factor; C080654:PCSK6 protein, human; D043484:Proprotein Convertases; D012697:Serine Endopeptidases; D045683:Furin
references.citation: Development. 2001 Nov;128(22):4501-10
references.pmid: 11714675
references.citation: J Cell Biol. 1999 Jul 26;146(2):345-59
references.pmid: 10465644
references.citation: J Biol Chem. 2008 Jan 25;283(4):2373-84
references.pmid: 18039650
references.citation: Biochem J. 2004 Mar 1;378(Pt 2):509-18
references.pmid: 14599293
references.citation: Development. 1993 Aug;118(4):1157-68
references.pmid: 7916676
references.citation: Nature. 1998 Oct 15;395(6703):702-7
references.pmid: 9790191
references.citation: Dev Cell. 2006 Sep;11(3):313-23
references.pmid: 16950123
references.citation: Curr Biol. 2005 Jan 11;15(1):31-6
references.pmid: 15649361
references.citation: PLoS One. 2008 Jan 16;3(1):e1434
references.pmid: 18197245
references.citation: EMBO J. 2002 Aug 1;21(15):3989-4000
references.pmid: 12145200
references.citation: Curr Biol. 2004 Apr 6;14(7):618-24
references.pmid: 15062104
references.citation: Cell. 2004 Sep 17;118(6):767-80
references.pmid: 15369675
references.citation: Nat Cell Biol. 2006 Jan;8(1):46-54
references.pmid: 16341206
references.citation: Plant Physiol. 2002 Nov;130(3):1102-8
references.pmid: 12427977
references.citation: Nat Cell Biol. 2002 Dec;4(12):981-5
references.pmid: 12447384
references.citation: Mol Biol Cell. 2006 Aug;17(8):3689-704
references.pmid: 16760436
references.citation: J Biol Chem. 2003 Feb 28;278(9):7564-72
references.pmid: 12482757
references.citation: Nat Cell Biol. 2003 May;5(5):410-21
references.pmid: 12717440
references.citation: J Biol Chem. 2003 Nov 28;278(48):48339-47
references.pmid: 12972410
references.citation: Dev Cell. 2006 Dec;11(6):763-74
references.pmid: 17141153
references.citation: Mol Biol Cell. 2003 Feb;14(2):417-31
references.pmid: 12589044
references.citation: Nat Rev Mol Cell Biol. 2005 Feb;6(2):181-7
references.pmid: 15688003
references.citation: EMBO J. 1994 May 15;13(10):2280-8
references.pmid: 8194519
references.citation: Biochem J. 2004 Oct 1;383(Pt 1):187-99
references.pmid: 15239668
references.citation: J Cell Biol. 1999 Jan 11;144(1):139-49
references.pmid: 9885250
references.citation: Mol Cell Biol. 2002 Jun;22(11):3905-26
references.pmid: 11997523
references.citation: Cell. 1999 Apr 2;97(1):121-32
references.pmid: 10199408
references.citation: Proc Natl Acad Sci U S A. 2004 Nov 2;101(44):15656-60
references.pmid: 15505202
references.citation: Development. 2007 Mar;134(6):1023-34
references.pmid: 17287255
references.citation: Mol Biol Cell. 2004 Aug;15(8):3542-52
references.pmid: 15146059
references.citation: J Biol Chem. 2002 Apr 12;277(15):12879-90
references.pmid: 11799113
references.citation: Mol Biol Cell. 2005 Nov;16(11):5215-26
references.pmid: 16135528
references.citation: Genes Dev. 2003 Jan 1;17(1):31-6
references.pmid: 12514096
references.citation: EMBO J. 1994 May 15;13(10):2305-12
references.pmid: 8194522
references.citation: Dev Cell. 2002 Apr;2(4):411-23
references.pmid: 11970892
references.citation: J Biol Chem. 1999 Mar 26;274(13):8624-9
references.pmid: 10085099
references.citation: Mol Biol Cell. 2001 Oct;12(10):3031-45
references.pmid: 11598189
references.citation: Development. 2006 Jul;133(13):2497-505
references.pmid: 16728477
references.citation: J Biol Chem. 2007 Dec 7;282(49):35772-86
references.pmid: 17925387
references.citation: J Biol Chem. 2002 Dec 13;277(50):48834-41
references.pmid: 12370178
references.citation: Development. 2006 Jan;133(2):319-29
references.pmid: 16368929
references.citation: J Biol Chem. 1995 Nov 3;270(44):26565-9
references.pmid: 7592877
references.citation: Nat Rev Mol Cell Biol. 2002 Oct;3(10):753-66
references.pmid: 12360192
references.citation: Mol Carcinog. 1996 Jan;15(1):44-56
references.pmid: 8561865
references.citation: Development. 2005 Dec;132(24):5539-51
references.pmid: 16291788
references.citation: Annu Rev Cell Dev Biol. 2003;19:589-621
references.pmid: 14570583
references.citation: Mol Cell Biol. 2002 Jul;22(13):4439-49
references.pmid: 12052855
references.citation: Nature. 2001 Jun 21;411(6840):965-9
references.pmid: 11418863
references.citation: Dev Biol. 2008 Mar 15;315(2):280-9
references.pmid: 18241853
references.citation: Genes Dev. 2001 Aug 1;15(15):2010-22
references.pmid: 11485994
references.citation: J Biol Chem. 2002 Dec 27;277(52):50333-40
references.pmid: 12386161