Biological activity of substrate-bound basic fibroblast growth factor (FGF2): recruitment of FGF receptor-1 in endothelial cell adhesion contacts.
Substrate-bound FGF2 promotes endothelial cell adhesion by interacting with alpha(v)beta(3) integrin. Here, endothelial GM7373 cells spread and organize focal adhesion plaques on immobilized FGF2, fibronectin (FN), and vitronectin (VN). alpha(v)beta(3) integrin, paxillin, focal adhesion kinase, vinculin and pp60(src) localize in cell-substratum contact sites on FGF2, FN or VN. However, only immobilized FGF2 induces a long-lasting activation of extracellular signal-regulated kinases(1/2) (ERK(1/2)) and cell proliferation that was inhibited by the ERK(1/2) inhibitor PD 098059 and the tyrosine kinase (TK) inhibitor tyrphostin 23, pointing to the engagement of FGF receptor (FGFR) at the basal side of the cell. To assess this hypothesis, GM7373 cells were transfected with a dominant negative TK(-)-DeltaFGFR1 mutant (GM7373-DeltaFGFR1 cells) or with the full-length receptor (GM7373-FGFR1 cells). Both transfectants adhere and spread on FGF2 but GM7373-DeltaFGFR1 cells do not proliferate. Also, parental and GM7373-FGFR1 cells, but not GM7373-DeltaFGFR1 cells, undergo morphological changes and increased motility on FGF2-coated plastic. Finally, FGFR1, but not TK(-)-DeltaFGFR1, localizes in cell adhesion contacts on immobilized FGF2. In conclusion, substrate-bound FGF2 induces endothelial cell proliferation, motility, and the recruitment of FGFR1 in cell-substratum contacts. This may contribute to the cross talk among intracellular signaling pathways activated by FGFR1 and alpha(v)beta(3) integrin in endothelial cells.
Tanghetti
Elena
E
Unit of General Pathology and Immunology, Department of Biomedical Sciences and Biotechnology, School of Medicine, University of Brescia, Via Valsabbina 19, 25123 Brescia, Italy.
Ria
Roberto
R
Dell'Era
Patrizia
P
Urbinati
Chiara
C
Rusnati
Marco
M
Ennas
Maria Grazia
MG
Presta
Marco
M
D000818:Animals; D001665:Binding Sites; D015153:Blotting, Western; D002417:Cattle; D002448:Cell Adhesion; D002455:Cell Division; D004305:Dose-Response Relationship, Drug; D004730:Endothelium, Vascular / Q000166:cytology*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D016222:Fibroblast Growth Factor 2 / Q000378:metabolism*; D005419:Flavonoids / Q000494:pharmacology; D005799:Genes, Dominant; D006801:Humans; D007150:Immunohistochemistry; D008855:Microscopy, Electron, Scanning; D010766:Phosphorylation; D010957:Plasmids / Q000378:metabolism; D011485:Protein Binding; D020794:Receptor Protein-Tyrosine Kinases / Q000737:chemistry* / Q000378:metabolism*; D051496:Receptor, Fibroblast Growth Factor, Type 1; D017468:Receptors, Fibroblast Growth Factor / Q000737:chemistry* / Q000378:metabolism*; D019030:Receptors, Vitronectin / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism; D015398:Signal Transduction; D013997:Time Factors; D014162:Transfection
D004791:Enzyme Inhibitors; D005419:Flavonoids; D017468:Receptors, Fibroblast Growth Factor; D019030:Receptors, Vitronectin; D011994:Recombinant Proteins; D016222:Fibroblast Growth Factor 2; C496348:FGFR1 protein, human; D020794:Receptor Protein-Tyrosine Kinases; D051496:Receptor, Fibroblast Growth Factor, Type 1; C093973:2-(2-amino-3-methoxyphenyl)-4H-1-benzopyran-4-one