Signal transduction through the fibronectin receptor induces collagenase and stromelysin gene expression.

Abstract

Abstract. We have investigated the effects of ligation of the fibronectin receptor (FnR) on gene expression in rabbit synovial fibroblasts. Monoclonal antibodies to the FnR that block initial adhesion of fibroblasts to fibronectin induced the expression of genes encoding the secreted extracellular matrix-degrading metalloproteinases collagenase and stromelysin. That induction was a direct consequence of interaction with the FnR was shown by the accumulation of mRNA for stromelysin and collagenase. Monoclonal antibodies to several other membrane glycoprotein receptors had no effect on metalloproteinase gene expression. Less than 2 h of treatment of the fibroblasts with anti-FnR in solution was sufficient to trigger the change in gene expression, and induction was blocked by dexamethasone. Unlike other inducers of metalloproteinase expression, including phorbol diesters and growth factors, addition of the anti-FnR in solution to cells adherent to serum-derived adhesion proteins or collagen produced no detectable change in cell shape or actin microfilament organization. Inductive effects were potentiated by cross-linking of the ligand. Fab fragments of anti-FnR were ineffective unless cross-linked or immobilized on the substrate. Adhesion of fibroblasts to native fibronectin did not induce metalloproteinases. However, adhesion to covalently immobilized peptides containing the arg-gly-asp sequence that were derived from fibronectin, varying in size from hexapeptides up to 120 kD, induced collagenase and stromelysin gene expression. This suggests that degradation products of fibronectin are the natural inductive ligands for the FnR. These data demonstrate that signals leading to changes in gene expression are transduced by the FnR, a member of the integrin family of extracellular matrix receptors. The signaling of changes in gene expression by the FnR is distinct from signaling involving cell shape and actin cytoarchitecture. At least two distinct signals are generated: the binding of fibronectin-derived fragments and adhesionblocking antibodies to the FnR triggers events different from those triggered by binding of the native fibronectin ligand. Because the genes regulated by this integrin are for enzymes that degrade the extracellular matrix, these results suggest that information transduced by the binding of various ligands to integrins may orchestrate the expression of genes regulating cell behavior in the extracellular environment.

Expression of stromelysin-3 in the human placenta and placental bed

“…This switch in adhesion-molecule expression has been shown to contribute to the acquisition of an invasive phenotype by cultured CTB (Damsky et al, 1992;Damsky, Sutherland and Fisher, 1993;Denker, 1993). Moreover, integrin-ligand interactions have been reported previously to modulate the expression of MMP in several cell lines (Werb et al, 1989;Seftor et al, 1993;Seltzer et al, 1994). Therefore, we hypothesize that during the differentiation of CTB into invasive IT, ST3 expression could be induced by specific integrin-mediated interactions with ECM components.…”

Stimulation of Matrix Metalloproteinase-9 Expression in Human Fibrosarcoma Cells by Synthetic Matrix Metalloproteinase Inhibitors

“…In this respect, several molecules, including cytokines such as IL-1β, TNF-α, epidermal growth factor, granulocyte macrophage-colony stimulating factor (GM-CSF), transforming growth factor-β, TNF-β, interferon-γ, IL-4, and IL-6 [52][53][54], as well as ECM components such as fibronectin, collagens, laminin, and tenascin [13,[55][56][57] have been reported as potent modulators of MMP-9 expression in different cell types. This raised the possibility that the effect of GI129471 on MMP-9 secretion might be mediated through modulations of the activities of these ECM components, cytokines, and/or their respective receptors.…”

β1-Integrin Cytoplasmic Subdomains Involved in Dominant Negative Function

“…The following antibodies were used: the rat anti-human ␤1 mAb 13 (Akiyama et al, 1989) was a gift from K. Yamada (National Institutes of Health, Bethesda, MD); the activating mouse anti-human ␤1 mAb TS2/16 (Hemler et al, 1984) was obtained from American Type Culture Collection (Rockville, MD); the mouse anti-human ␤1 mAb 12G10 was characterized previously (Mould et al, 1995); the rat anti-mouse ␤1 mAb 9EG7, with human cross-reactivity (Lenter et al, 1993), was a gift from D. Vestweber (ZMBE Technologiehof, Muenster, Germany); the blocking mouse anti-human ␤1 mAb AIIB2 (Werb et al, 1989) was a gift from C. Damsky (Department of Stomatology, University of California, San Francisco, CA); the inhibitory PB1 mAb against hamster ␣5␤1 heterodimer was obtained from the Developmental Studies Hybridoma Bank (University of Iowa, Iowa City, IA); the blocking anti-mouse ␣V H9.2B8 mAb (Moulder et al, 1991) was purchased from PharMingen (San Diego, CA); the rat anti-mouse ␣6 mAb GoH3 (Sonnenberg et al, 1988) was a gift from A. Sonnenberg (The Neederland Cancer Institute, Amsterdam, the Netherlands); the anti-talin mAb 8d4 was obtained S.F. Retta et al…”

“…The following antibodies were used: the rat anti-human ␤1 mAb 13 (Akiyama et al, 1989) was a gift from K. Yamada (National Institutes of Health, Bethesda, MD); the activating mouse anti-human ␤1 mAb TS2/16 (Hemler et al, 1984) was obtained from American Type Culture Collection (Rockville, MD); the mouse anti-human ␤1 mAb 12G10 was characterized previously (Mould et al, 1995); the rat anti-mouse ␤1 mAb 9EG7, with human cross-reactivity (Lenter et al, 1993), was a gift from D. Vestweber (ZMBE Technologiehof, Muenster, Germany); the blocking mouse anti-human ␤1 mAb AIIB2 (Werb et al, 1989) was a gift from C. Damsky (Department of Stomatology, University of California, San Francisco, CA); the inhibitory PB1 mAb against hamster ␣5␤1 heterodimer was obtained from the Developmental Studies Hybridoma Bank (University of Iowa, Iowa City, IA); the blocking anti-mouse ␣V H9.2B8 mAb (Moulder et al, 1991) was purchased from PharMingen (San Diego, CA); the rat anti-mouse ␣6 mAb GoH3 (Sonnenberg et al, 1988) was a gift from A. Sonnenberg (The Neederland Cancer Institute, Amsterdam, the Netherlands); the anti-talin mAb 8d4 was obtained S.F. Retta et al from Sigma (St. Louis, MO); the anti ␣-actinin mAb 1682 was from Chemicon (Temecula, CA); rabbit polyclonal antisera to human fibronectin and to ␣V, ␣3, and ␣5 integrin cytoplasmic domains were produced in our laboratory (Tarone et al, 1984, Defilippi et al, 1992; the FAK 4 polyclonal antibody and the mAb FAK9.2 for FAK immunoprecipitation and Western blotting, respectively, were prepared in our laboratory as described previously (Defilippi et al, 1995); a mouse anti-paxillin mAb and the anti-phosphotyrosine mAb PY20 were purchased from Transduction Laboratories (Nottingham, UK); fluorescein-labeled phalloidin, fluorescein-labeled goat anti-mouse IgG, rhodamine-labeled goat anti-mouse IgG, and rhodamine-labeled goat anti-rabbit IgG were all from Sigma.…”