Fibronectin's cell-adhesive domain and an amino-terminal matrix assembly domain participate in its assembly into fibroblast pericellular matrix.

McDonald, John A.; Quade, Bradley J.; Broekelmann, T J; Lachance, Rhonda M.; Forsman, K; Hasegawa, Eiichi; Akiyama, Steven K.

doi:10.1016/s0021-9258(18)61453-x

Cited by 236 publications

(49 citation statements)

References 62 publications

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“…This initial binding to the cell adhesion domain is rapid but of moderate affinity that has been reported for binding of soluble fibronectin to ceils in suspension (Akiyama and Yamada, 1984). Such a moderate affinity interaction might explain why no direct binding of iodinated fibronectin cell adhesion fragments could be measured (McKeown-Longo and Mosher, 1985) and why large amounts of unlabeled cell adhesion fragments are required to compete fibronectin binding and matrix assembly (McDonald et al, 1987; our unpublished resultsg. The anti-Bi would therefore block binding of soluble fibronectin to cell layers and its subsequent incorporation into the matrix by interfering with the initial transient binding event before the unfolding of the amino-terminal 70-kD domain.…”

Section: Discussionmentioning

confidence: 52%

“…First, 70-and 27-kD amino-terminal fragments, lacking the RGDS cell attachment site, bind to fibroblast monolayers and compete with exogenous fibronectin for binding (McKeown-Longo and Mosher, 1985;Quade and McDonald, 1988). Second, the 70-kD amino-terminal fragment (70 kD) and a monoclonal antibody to the fibronectin aminoterminal region block endogenously synthesized fibronectin matrix assembly (McDonald et al, 1987). Third, 3T3 cells, infected with a retroviral expression vector containing cDNA that encodes for fibronectin lacking the amino-terminal half, synthesize and secrete recombinant truncated fibronectins referred to as deminectins.…”

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confidence: 99%

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Inhibition of binding of fibronectin to matrix assembly sites by anti-integrin (alpha 5 beta 1) antibodies.

Fogerty

Akiyama

Yamada

et al. 1990

The Journal of Cell Biology

219

150

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Abstract. Fibroblasts have cell surface sites that mediate assembly of plasma and cellular fibronectin into the extracellular matrix. Cell adhesion to fibronectin can be mediated by the interaction of an integrin (oL5/3~) with the Arg-Gly-Asp-Ser (RGDS)-containing cell adhesion region of fibronectin. We have attempted to elucidate the role of the otsfll fibronectin receptor in assembly of fibronectin in matrices. Rat monoclonal antibody mAb 13, which recognizes the integrin fll subunit, completely blocked binding and matrix assembly of 125I-fibronectin as well as binding of the 1251-70-kD amino-terminal fragment of fibronectin (70 kD) to fibroblast cell layers. Fab fragments of the anti-/~ antibody were also inhibitory. Antibody mAb 16, which recognizes the integrin ors subunit, partially blocked binding of 125I-fibronectin and 12q-70-kD.When cell layers were coincubated with fluoresceinated fibronectin and either anti-13~ or anti-us, anti-131 was a more effective inhibitor than anti-o~s of binding of labeled fibronectin to the cell layer. Inhibition of ~2q-fibmnectin binding by anti-/31 IgG occurred within 20 min. Inhibition of 125I-fibronectin binding by anti-/3t Fab fragments or IgG could not be overcome with increasing concentrations of fibronectin, suggesting that anti-131 and exogenous fibronectin may not compete for the same binding site. No/31-containing integrin bound to immobilized 70 kD. These data indicate that the/31 subunit plays an important role in binding and assembly of exogenous fibronectin, perhaps by participation in the organization, regeneration, or cycling of the assembly site rather than by a direct interaction with fibronectin.

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Section: Discussionmentioning

confidence: 52%

mentioning

confidence: 99%

Inhibition of binding of fibronectin to matrix assembly sites by anti-integrin (alpha 5 beta 1) antibodies.

Fogerty

Akiyama

Yamada

et al. 1990

The Journal of Cell Biology

219

150

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“…Murine laminin and rabbit anti-mouse laminin IgG were purchased from Collaborative Research, Inc. (Waltham, MA) and Bethesda Research Laboratories (Gaithersburg, MD), respectively. The derivation, isolation, and characterization of mAb 333 (5) and mAb 304 (45) have been described previously, mAb 333 binds close to the Arg-Gly-Asp-Ser adhesive recognition signal in the central cell-binding domain of fibronectin, and is a potent inhibitor of both the direct binding of fibronectin to fibroblastic cells and of fibroblastic cell adhesion (5,29), while mAb 304 binds specifically to the NH~-terminal heparin/fibrin-binding domain of fibronectin (45). The synthesis, purification, and analysis of synthetic peptides spanning the IIICS region of human fibronectin (connecting segment [CS] peptides) were performed as described previously (29).…”

Section: Methodsmentioning

confidence: 99%

Neurite extension of chicken peripheral nervous system neurons on fibronectin: relative importance of specific adhesion sites in the central cell-binding domain and the alternatively spliced type III connecting segment.

Humphries

Akiyama

Komoriya

et al. 1988

The Journal of Cell Biology

128

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Abstract. Fibronectin contains at least two domainsthat support cell adhesion. One is the central cellbinding domain that is recognized by a variety of cell types, including fibroblasts. The second, originally identified by its ability to support melanoma cell adhesion, is located in the alternatively spliced type III connecting segment (IIICS). Using specific adhesive ligands and inhibitory probes, we have examined the role of each of these domains in fibronectin-mediated neurite extension of neurons from chick embryo dorsal root and sympathetic ganglia. In studies using explanted ganglia, both f13, a 75-kD tryptic fragment of human plasma fibronectin containing the central cellbinding domain, and CSI-IgG, a synthetic peptide-IgG conjugate containing the principal cell adhesion site from the IIICS, supported neurite outgrowth after adsorption onto the substrate. The maximal activities of f13 and CSI-IgG were 45-55% and 25-30% that of intact fibronectin, respectively. Co-coating of the substrate with f13 and CSI-IgG produced an additive stimulation of neurite outgrowth, the extent of which approached that obtained with fibronectin. Similar results were obtained with purified neuronal cell preparations isolated by tryptic dissociation of dorsal root ganglia. In complementary studies, blockage of the adhesive function of either the central cell-binding domain (with mAb 333, an antiadhesive monoclonal antibody) or the IIICS (with CS1 peptide), resulted in ~60 or 30% reduction in fibronectin-mediated neurite outgrowth, respectively. When tested in combination, the inhibitory activities of mAb 333 and CS1 were additive. From these results, we conclude that neurons from the peripheral nervous system can extend neurites on both the central cell-binding domain and the IIICS region of fibronectin, and that these cells are therefore the first normal, embryonic cell type shown to adhere to the IIICS. These results suggest that spatiotemporal fluctuations in the alternative mRNA splicing of the IIICS region of fibronectin may be important in regulation of cell adhesive events during development of the peripheral nervous system. URING development of the embryonic nervous system and regeneration of damaged adult nerve fibers, innervation patterns are established via specific pathways (19,65). The interaction of neuronal growth cones with their environment is considered to be of central importance for directing axons to their target sites. Thus, one of the principal goals in neuroscience is elucidation of the mechanisms that regulate axonal guidance.The substrates encountered by elongating axons are a complex mixture of neuronal and nonneuronal cell surfaces, axonal processes, and fibronectin-and laminin-containing extracellular matrices, each of which has been implicated in the directed elongation of neuronal cells in vivo (7,12,32,55,56,63,66,75). During development of the peripheral nervous system (PNS) ~, neuronal precursor cells originating from the neural crest travel long distances through intersomitic spaces that are r...

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“…Interestingly, anti-α2 treatment reduced osteogenesis to a lesser extent than HFN7.1. Not only does HFN7.1 abrogate cell adhesion to fibronectin, but perturbation of α5β1 and αvβ3 integrin recognition also alters fibronectin conformation and fibril formation 36,37 , further enhancing its effects. This is also consistent with the observation that sol RGD peptides compete against the cell-adhesive domain of fibronectin for interactions with integrins, and thus similarly reduces hMSC osteogenic differentiation (Fig.…”

Section: Nascent Protein Adhesion Influences Cell Mechanosensingmentioning

confidence: 99%

Local nascent protein deposition and remodelling guide mesenchymal stromal cell mechanosensing and fate in three-dimensional hydrogels

2019

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Hydrogels serve as valuable tools for studying cell-extracellular matrix (ECM) interactions in three-dimensional (3D) environments that recapitulate aspects of native ECM. However, the impact of early protein deposition on cell behavior within hydrogels has largely been overlooked. Using a bio-orthogonal labeling technique, we visualized nascent proteins within a day of culture across a range of hydrogels. In two engineered hydrogels of interest in 3D mechanobiology studies – proteolytically degradable covalently crosslinked hyaluronic acid (HA) and dynamic viscoelastic HA hydrogels – mesenchymal stromal cell (MSC) spreading, YAP/TAZ nuclear translocation, and osteogenic differentiation were observed with culture. However, inhibition of cellular adhesion to nascent proteins or reduction in nascent protein remodeling reduced MSC spreading and nuclear translocation of YAP/TAZ, resulting in a shift towards adipogenic differentiation. Our findings emphasize the role of nascent proteins in the cellular perception of engineered materials and have implications for in vitro cell signaling studies and application to tissue repair.

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Fibronectin's cell-adhesive domain and an amino-terminal matrix assembly domain participate in its assembly into fibroblast pericellular matrix.

Cited by 236 publications

References 62 publications

Inhibition of binding of fibronectin to matrix assembly sites by anti-integrin (alpha 5 beta 1) antibodies.

Inhibition of binding of fibronectin to matrix assembly sites by anti-integrin (alpha 5 beta 1) antibodies.

Neurite extension of chicken peripheral nervous system neurons on fibronectin: relative importance of specific adhesion sites in the central cell-binding domain and the alternatively spliced type III connecting segment.

Local nascent protein deposition and remodelling guide mesenchymal stromal cell mechanosensing and fate in three-dimensional hydrogels

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