Enhancement of Notch receptor maturation and signaling sensitivity by Cripto-1.
Nodal and Notch signaling pathways play essential roles in vertebrate development. Through a yeast two-hybrid screening, we identified Notch3 as a candidate binding partner of the Nodal coreceptor Cripto-1. Coimmunoprecipitation analysis confirmed the binding of Cripto-1 with all four mammalian Notch receptors. Deletion analyses revealed that the binding of Cripto-1 and Notch1 is mediated by the Cripto-1/FRL-1/Cryptic domain of Cripto-1 and the C-terminal region of epidermal growth factor-like repeats of Notch1. Binding of Cripto-1 to Notch1 occurred mainly in the endoplasmic reticulum-Golgi network. Cripto-1 expression resulted in the recruitment of Notch1 protein into lipid raft microdomains and enhancement of the furin-like protein convertase-mediated proteolytic maturation of Notch1 (S1 cleavage). Enhanced S1 cleavage resulted in the sensitization to ligand-induced activation of Notch signaling. In addition, knockdown of Cripto-1 expression in human and mouse embryonal carcinoma cells desensitized the ligand-induced Notch signaling activation. These results suggest a novel role of Cripto-1 in facilitating the posttranslational maturation of Notch receptors.
Watanabe
Kazuhide
K
Mammary Biology and Tumorigenesis Laboratory, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892, USA.
Nagaoka
Tadahiro
T
Lee
Joseph M
JM
Bianco
Caterina
C
Gonzales
Monica
M
Castro
Nadia P
NP
Rangel
Maria Cristina
MC
Sakamoto
Kei
K
Sun
Youping
Y
Callahan
Robert
R
Salomon
David S
DS
D020816:Amino Acid Motifs; D000818:Animals; D001665:Binding Sites; D016466:CHO Cells; D019556:COS Cells; D002478:Cells, Cultured; D002522:Chlorocebus aethiops; D006224:Cricetinae; D003412:Cricetulus; D004815:Epidermal Growth Factor / Q000737:chemistry / Q000378:metabolism*; D016326:Extracellular Matrix Proteins / Q000378:metabolism; D058851:GPI-Linked Proteins; D015723:Gene Library; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins; D008562:Membrane Glycoproteins / Q000737:chemistry / Q000378:metabolism*; D021962:Membrane Microdomains / Q000378:metabolism; D051379:Mice; D009363:Neoplasm Proteins / Q000737:chemistry / Q000378:metabolism*; D025941:Protein Interaction Mapping; D051881:Receptor, Notch1 / Q000737:chemistry / Q000378:metabolism*; D000071656:Receptor, Notch3; D051880:Receptors, Notch / Q000737:chemistry / Q000378:metabolism; D015398:Signal Transduction; D020798:Two-Hybrid System Techniques
C485883:Efemp2 protein, mouse; D016326:Extracellular Matrix Proteins; D058851:GPI-Linked Proteins; D036341:Intercellular Signaling Peptides and Proteins; D008562:Membrane Glycoproteins; D009363:Neoplasm Proteins; C499373:Notch3 protein, mouse; D051881:Receptor, Notch1; D000071656:Receptor, Notch3; D051880:Receptors, Notch; C070636:TDGF1 protein, human; D004815:Epidermal Growth Factor