Mechanism of Talin-dependent control of avb3 integrin-mediated adhesion and signaling

Les protéines de la famille des intégrines sont impliquées dans le lien physique qui s'établit entre les protéines de la matrice extracellulaire et le cytosquelette d'actine lorsque les cellules adhèrent à la matrice environnante. Les intégrines sont des récepteurs transmembranaires qui existent dans une forme active ou inactive à la surface des cellules. La voie intracellulaire d'activation des intégrines et le regroupement de ces récepteurs transmembranaires dans les zones d'adhésion focales sont deux mécanismes induits par la taline, protéine adaptatrice intracellulaire. L'expression transitoire d'une forme sauvage (WT), ou mutée dans la partie cytoplasmique de la chaîne b3 taggée avec une protéine fluorescente, dans des cellules n'exprimant pas d'intégrine b3 de façon endogène, nous a permis d'analyser le rôle de l'interaction intégrine b3-taline lors de l'adhésion des cellules transfectées sur un substrat spécifique des intégrines de type b3 telle que la vitronectine

Integrins: versatile receptors controlling melanocyte adhesion, migration and proliferation

From the onset of melanocyte specification from the neural crest, throughout their migration during embryogenesis and until they reside in their niche in the basal keratinocyte layer, melanocytes interact in dynamic ways with the extracellular environment of the growing embryo. To recognize and to adhere to their environment, melanocytes depend on heterodimeric cell surface receptors of the family of integrins. In addition to the control of adhesive interactions between melanocytes and the extracellular matrix scaffold secreted by fibroblasts and keratinocytes, the integrin receptors allow cells also to sense the mechanical condition of the extracellular environment, responding by intracellular signaling, triggering cell survival, proliferation or migration events. In this review, we summarize the recently emerged concepts that explain integrin-dependent adhesion and how this adhesion system interfaces with integrin-dependent signaling events. The gained information will help to understand melanocyte behavior in pathological situations such as melanoma growth and metastasis formation

Niche anchorage and signaling through membrane-bound Kit-ligand/c-kit receptor are kinase independent and imatinib insensitive

Kit ligand (KitL) and its tyrosine kinase receptor c-kit are critical for germ cells, melanocytes, mastocytes, and hematopoietic stem cells. Alternative splicing of KitL generates membrane-bound KitL (mb-KitL) or soluble KitL, providing survival or cell migration, respectively. Here we analyzed whether c-kit can function both as an adhesion and signaling receptor to mb-KitL presented by the environmental niche. At contacts between fibroblasts and MC/9 mast cells, mb-KitL, and c-kit formed ligand/receptor clusters that formed stable complexes, which resisted dissociation by c-kit blocking mAbs and provided cell anchorage under physiological shear stresses. Clusters recruited tyrosine-phosphorylated proteins and induced spatially restricted F-actin polymerization. Mutational analysis of c-kit demonstrated kinase-independent mb-KitL/c-kit clustering, anchorage, F-actin polymerization, and Tyr567-dependent cluster phosphorylation. Kinase inhibition of c-kit by imatinib reduced cluster coalescence, but allowed cluster phosphorylation and F-actin polymerization, which required PI3K recruitment and a newly identified juxtamembrane residue. Synergies between integrin and c-kit-mediated spreading and adhesion of MC/9 cells were studied in vitro on immobilized-KitL/fibronectin surfaces. While c-kit blocking antibodies prevented spreading, imatinib blocked spreading induced by soluble- but not immobilized KitL. Thus, "mechanical" activation of c-kit provides signaling, niche-anchorage, and synergies with integrin-mediated adhesion, which is independent of kinase function and resistant to c-kit kinase inhibitors.

Talin-bound NPLY motif recruits integrin-signaling adapters to regulate cell spreading and mechanosensing

Integrin-dependent cell adhesion and spreading are critical for morphogenesis, tissue regeneration, and immune defense but also tumor growth. However, the mechanisms that induce integrin-mediated cell spreading and provide mechanosensing on different extracellular matrix conditions are not fully understood. By expressing β3-GFP-integrins with enhanced talin-binding affinity, we experimentally uncoupled integrin activation, clustering, and substrate binding from its function in cell spreading. Mutational analysis revealed Tyr747, located in the first cytoplasmic NPLY(747) motif, to induce spreading and paxillin adapter recruitment to substrate- and talin-bound integrins. In addition, integrin-mediated spreading, but not focal adhesion localization, was affected by mutating adjacent sequence motifs known to be involved in kindlin binding. On soft, spreading-repellent fibronectin substrates, high-affinity talin-binding integrins formed adhesions, but normal spreading was only possible with integrins competent to recruit the signaling adapter protein paxillin. This proposes that integrin-dependent cell-matrix adhesion and cell spreading are independently controlled, offering new therapeutic strategies to modify cell behavior in normal and pathological conditions