Requirement of glycosylphosphatidylinositol anchor of Cripto-1 for trans activity as a Nodal co-receptor.
Cripto-1 (CR-1) has an indispensable role as a Nodal co-receptor for patterning of body axis in embryonic development. CR-1 is reported to have a paracrine activity as a Nodal co-receptor, although CR-1 is primarily produced as a glycosylphosphatidylinositol (GPI)-anchored membrane protein. Regulation of cis and trans function of CR-1 should be important to establish the precise body patterning. However, the mechanism by which GPI-anchored CR-1 can act in trans is not well known. Here we confirmed the paracrine activity of CR-1 by fluorescent cell-labeling and immunofluorescent staining. We generated COOH-terminal-truncated soluble forms of CR-1 based on the attachment site for the GPI moiety (omega-site), which we identified in the present study. GPI-anchored CR-1 has a significantly higher activity than COOH-terminal-truncated soluble forms to induce Nodal signal in trans as well as in cis. Moreover, transmembrane forms of CR-1 partially retained their ability to induce Nodal signaling only when type I receptor Activin-like kinase 4 was overexpressed. NTERA2/D1 cells, which express endogenous CR-1, lost the cell-surface expression of CR-1 after phosphatidylinositol-phospholipase C treatment and became refractory to stimulation of Nodal. These observations suggest that GPI attachment of CR-1 is required for the paracrine activity as a Nodal co-receptor.
Watanabe
Kazuhide
K
Tumor Growth Factor Section, Mammary Biology and Tumorigenesis Laboratory, Center for Cancer Research, NCI, National Institutes of Health, Bethesda, Maryland 20892, USA.
Hamada
Shin
S
Bianco
Caterina
C
Mancino
Mario
M
Nagaoka
Tadahiro
T
Gonzales
Monica
M
Bailly
Veronique
V
Strizzi
Luigi
L
Salomon
David S
DS
D030201:Activin Receptors, Type I / Q000378:metabolism; D019521:Body Patterning / Q000502:physiology; D002460:Cell Line; D047108:Embryonic Development / Q000502:physiology; D004815:Epidermal Growth Factor / Q000378:metabolism*; D058851:GPI-Linked Proteins; D017261:Glycosylphosphatidylinositols / Q000378:metabolism*; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins; D008562:Membrane Glycoproteins / Q000378:metabolism*; D009363:Neoplasm Proteins / Q000378:metabolism*; D055457:Nodal Protein; D019899:Paracrine Communication / Q000187:drug effects / Q000502:physiology*; D054801:Phosphoinositide Phospholipase C / Q000494:pharmacology; D020033:Protein Isoforms / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects / Q000502:physiology*; D016212:Transforming Growth Factor beta / Q000378:metabolism*
D058851:GPI-Linked Proteins; D017261:Glycosylphosphatidylinositols; D036341:Intercellular Signaling Peptides and Proteins; D008562:Membrane Glycoproteins; C526007:NODAL protein, human; D009363:Neoplasm Proteins; D055457:Nodal Protein; D020033:Protein Isoforms; C070636:TDGF1 protein, human; D016212:Transforming Growth Factor beta; D004815:Epidermal Growth Factor; D030201:Activin Receptors, Type I; D054801:Phosphoinositide Phospholipase C