PMID:14625290

Citation	Cruz, AC, Frank, BT, Edwards, ST, Dazin, PF, Peschon, JJ and Fang, KC (2004) Tumor necrosis factor-alpha-converting enzyme controls surface expression of c-Kit and survival of embryonic stem cell-derived mast cells. J. Biol. Chem. 279:5612-20
Abstract	Transmembrane metalloproteinases of the disintegrin and metalloproteinase (ADAM) family control cell signaling interactions via hydrolysis of protein extracellular domains. Prior work has shown that the receptor tyrosine kinase, c-Kit (CD117), is essential for mast cell survival and that serum levels of c-Kit increase in proliferative mast cell disorders, suggesting the existence of c-Kit shedding pathways in mast cells. In the present work, we report that tumor necrosis factor alpha-converting enzyme (TACE; ADAM-17) mediates shedding of c-Kit. Stimulation of transfected cells with phorbol 12-myristate 13-acetate (PMA) induced metalloproteinase-mediated release of c-Kit ectodomain, which increased further upon TACE overexpression. By contrast, TACE-deficient fibroblasts did not demonstrate inducible release, thus identifying TACE as the metalloproteinase primarily responsible for PMA-induced c-Kit shedding. Surface expression of c-Kit by the human mast cell-1 line decreased upon phorbol-induced shedding, which involved metalloproteinase activity susceptible to inhibition by tissue inhibitor of metalloproteinase (TIMP)-3. To further explore the role of TACE in shedding of c-Kit from mast cells, we compared the behavior of mast cells derived from murine embryonic stem cells. In these studies, PMA decreased surface c-Kit levels on mast cells expressing wild-type (+/+) TACE but not on those expressing an inactive mutant (DeltaZn/DeltaZn), confirming the role of TACE in PMA-induced c-Kit shedding. Compared with TACE(+/+) cells, TACE(DeltaZn/DeltaZn) mast cells also demonstrated decreased constitutive shedding and increased basal surface expression of c-Kit, with diminished apoptosis in response to c-Kit ligand deprivation. These data suggest that TACE controls mast cell survival by regulating shedding and surface expression of c-Kit.
Links	PubMed Online version:10.1074/jbc.M312323200
Keywords	ADAM Proteins; Animals; Apoptosis; Cell Differentiation; Cell Division; Cell Line; Cell Separation; Cell Survival; Cytoplasm/metabolism; Detergents/pharmacology; Dose-Response Relationship, Drug; Embryo, Mammalian/cytology; Fibroblasts/metabolism; Flow Cytometry; Humans; Immunoblotting; Ligands; Mast Cells/metabolism; Metalloendopeptidases/metabolism; Mice; Mice, Transgenic; Mutagenesis, Site-Directed; Polyethylene Glycols/pharmacology; Precipitin Tests; Protein Structure, Tertiary; Proto-Oncogene Proteins c-kit/biosynthesis; Proto-Oncogene Proteins c-kit/metabolism; Stem Cell Factor/metabolism; Stem Cells/metabolism; Transfection
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