Cripto recruits Furin and PACE4 and controls Nodal trafficking during proteolytic maturation.
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.
Blanchet
Marie-Hélène
MH
Ecole Polytechnique Fédérale de Lausanne (EPFL), Swiss Institute for Experimental Cancer Research (ISREC), Epalinges, Switzerland.
Le Good
J Ann
JA
Mesnard
Daniel
D
Oorschot
Viola
V
Baflast
Stéphane
S
Minchiotti
Gabriella
G
Klumperman
Judith
J
Constam
Daniel B
DB
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
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
Development. 2001 Nov;128(22):4501-10
11714675
J Cell Biol. 1999 Jul 26;146(2):345-59
10465644
J Biol Chem. 2008 Jan 25;283(4):2373-84
18039650
Biochem J. 2004 Mar 1;378(Pt 2):509-18
14599293
Development. 1993 Aug;118(4):1157-68
7916676
Nature. 1998 Oct 15;395(6703):702-7
9790191
Dev Cell. 2006 Sep;11(3):313-23
16950123
Curr Biol. 2005 Jan 11;15(1):31-6
15649361
PLoS One. 2008 Jan 16;3(1):e1434
18197245
EMBO J. 2002 Aug 1;21(15):3989-4000
12145200
Curr Biol. 2004 Apr 6;14(7):618-24
15062104
Cell. 2004 Sep 17;118(6):767-80
15369675
Nat Cell Biol. 2006 Jan;8(1):46-54
16341206
Plant Physiol. 2002 Nov;130(3):1102-8
12427977
Nat Cell Biol. 2002 Dec;4(12):981-5
12447384
Mol Biol Cell. 2006 Aug;17(8):3689-704
16760436
J Biol Chem. 2003 Feb 28;278(9):7564-72
12482757
Nat Cell Biol. 2003 May;5(5):410-21
12717440
J Biol Chem. 2003 Nov 28;278(48):48339-47
12972410
Dev Cell. 2006 Dec;11(6):763-74
17141153
Mol Biol Cell. 2003 Feb;14(2):417-31
12589044
Nat Rev Mol Cell Biol. 2005 Feb;6(2):181-7
15688003
EMBO J. 1994 May 15;13(10):2280-8
8194519
Biochem J. 2004 Oct 1;383(Pt 1):187-99
15239668
J Cell Biol. 1999 Jan 11;144(1):139-49
9885250
Mol Cell Biol. 2002 Jun;22(11):3905-26
11997523
Cell. 1999 Apr 2;97(1):121-32
10199408
Proc Natl Acad Sci U S A. 2004 Nov 2;101(44):15656-60
15505202
Development. 2007 Mar;134(6):1023-34
17287255
Mol Biol Cell. 2004 Aug;15(8):3542-52
15146059
J Biol Chem. 2002 Apr 12;277(15):12879-90
11799113
Mol Biol Cell. 2005 Nov;16(11):5215-26
16135528
Genes Dev. 2003 Jan 1;17(1):31-6
12514096
EMBO J. 1994 May 15;13(10):2305-12
8194522
Dev Cell. 2002 Apr;2(4):411-23
11970892
J Biol Chem. 1999 Mar 26;274(13):8624-9
10085099
Mol Biol Cell. 2001 Oct;12(10):3031-45
11598189
Development. 2006 Jul;133(13):2497-505
16728477
J Biol Chem. 2007 Dec 7;282(49):35772-86
17925387
J Biol Chem. 2002 Dec 13;277(50):48834-41
12370178
Development. 2006 Jan;133(2):319-29
16368929
J Biol Chem. 1995 Nov 3;270(44):26565-9
7592877
Nat Rev Mol Cell Biol. 2002 Oct;3(10):753-66
12360192
Mol Carcinog. 1996 Jan;15(1):44-56
8561865
Development. 2005 Dec;132(24):5539-51
16291788
Annu Rev Cell Dev Biol. 2003;19:589-621
14570583
Mol Cell Biol. 2002 Jul;22(13):4439-49
12052855
Nature. 2001 Jun 21;411(6840):965-9
11418863
Dev Biol. 2008 Mar 15;315(2):280-9
18241853
Genes Dev. 2001 Aug 1;15(15):2010-22
11485994
J Biol Chem. 2002 Dec 27;277(52):50333-40
12386161