c-Kit receptor signaling through its phosphatidylinositide-3'-kinase- binding site and protein kinase C: Role in mast cell enhancement of degranulation, adhesion, and membrane ruffling Journal Article
| Authors: | Vosseller, K.; Stella, G.; Yee, N. S.; Besmer, P. |
| Article Title: | c-Kit receptor signaling through its phosphatidylinositide-3'-kinase- binding site and protein kinase C: Role in mast cell enhancement of degranulation, adhesion, and membrane ruffling |
| Abstract: | In bone marrow-derived mast cells (BMMCs), the Kit receptor tyrosine kinase mediates diverse responses including proliferation, survival, chemotaxis, migration, differentiation, and adhesion to extracellular matrix. In connective tissue mast cells, a role for Kit in the secretion of inflammatory mediators has been demonstrated as well. We recently demonstrated a role for phosphatidylinositide-3' (PI 3)-kinase in Kit-ligand (KL)-induced adhesion of BMMCs to fibronectin. Herein, we investigated the mechanism by which Kit mediates enhancement of FcεRI-mediated degranulation, cytoskeletal rearrangements, and adhesion in BMMCs. W(sh)/W(sh) BMMCs, lacking endogenous Kit expression, were transduced to express normal and mutant Kit receptors containing Tyr → Phe substitutions at residues 719 and 821. Although the normal Kit receptor fully restored KL-induced responses in W(sh)/W(sh) BMMCs, Kit(Y719F), which fails to bind and activate PI 3-kinase, failed to potentiate degranulation and is impaired in mediating membrane ruffling and actin assembly. Inhibition of PI 3-kinase with wortmannin or LY294002 also inhibited secretory enhancement and cytoskeletal rearrangements mediated by Kit. In contrast, secretory enhancement and adhesion stimulated directly through protein kinase C (PKC) do not require PI 3-kinase. Calphostin C, an inhibitor of PKC, blocked Kit-mediated adhesion to fibronectin, secretory enhancement, membrane ruffling, and filamentous actin assembly. Although cytochalasin D inhibited Kit-mediated filamentous actin assembly and membrane ruffling, secretory enhancement and adhesion to fibronectin were not affected by this drug. Therefore, Kit-mediated cytoskeletal rearrangements that are dependent on actin polymerization can be uncoupled from the Kit-mediated secretory and adhesive responses. Our results implicate receptor-proximal PI 3-kinase activation and activation of a PKC isoform in Kit-mediated secretory enhancement, adhesion, and cytoskeletal reorganization. |
| Keywords: | signal transduction; controlled study; nonhuman; mutant protein; animal cell; mouse; animals; mice; actin; stem cell factor receptor; proto-oncogene proteins c-kit; amino acid substitution; protein assembly; protein protein interaction; protein binding; phosphatidylinositol 3 kinase; mice, inbred c57bl; animalia; protein-serine-threonine kinases; phosphotransferases (alcohol group acceptor); cell membrane; 1-phosphatidylinositol 3-kinase; binding site; protein kinase c; binding sites; up-regulation; actins; cell adhesion; cytoskeleton; calcium cell level; isoenzymes; serotonin; mast cells; calcium ion; membrane structure; ribosomal protein s6 kinases; fibronectins; cell degranulation; priority journal; article; mast cell degranulation; phytohemagglutinins |
| Journal Title: | Molecular Biology of the Cell |
| Volume: | 8 |
| Issue: | 5 |
| ISSN: | 1059-1524 |
| Publisher: | The American Society for Cell Biology |
| Date Published: | 1997-05-01 |
| Start Page: | 909 |
| End Page: | 922 |
| Language: | English |
| PUBMED: | 9168474 |
| PROVIDER: | scopus |
| PMCID: | PMC276137 |
| DOI/URL: | |
| Notes: | Article -- Export Date: 17 March 2017 -- Source: Scopus |
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