RhoA activity is required for fibronectin assembly and counteracts beta1B integrin inhibitory effect in FRT epithelial cells.
FRT thyroid epithelial cells synthesize fibronectin and organize a network of fibronectin fibrils at the basal surface of the cells. Fibronectin fibril formation is enhanced by the overexpression of the ubiquitous beta1A integrin and is inhibited by the expression of the dominant-negative beta1B subunit. We tested the hypotheses that RhoA activity might mediate the integrin-dependent fibronectin fibrillogenesis and might counteract beta1B integrin inhibitory effect. FRT-beta1A cells were transfected with a vector carrying a dominant negative form of RhoA (RhoAN19) or treated with the C3 transferase exoenzyme. Both treatments inhibited fibronectin assembly and caused loss of actin microfilaments and adhesion plaques. On the other hand, FRT-beta1B cells were transfected with the constitutively activated form of RhoA (RhoAV14) or treated with the E. coli cytotoxic necrotizing factor 1, which directly activates RhoA. Either treatment restored microfilament and adhesion plaque assembly and promoted fibronectin fibril organization. A great increase in fibronectin fibril assembly was also obtained by treatment of FRT-beta1B cells with TGF-beta. Our data indicate that RhoA is required to promote fibronectin matrix assembly in FRT cells and that the activation of the signal transduction pathway downstream of RhoA can overcome the inhibitory effect of beta1B integrin.
Centro di Endocrinologia ed Oncologia Sperimentale del CNR - Dipartimento di Biologia e Patologia Cellulare e Molecolare, Università degli Studi di Napoli 'Federico II', Napoli, Italy.
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Calì
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Mazzarella
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Chiacchio
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Negri
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Retta
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Zannini
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Gentile
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Tarone
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Nitsch
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Garbi
D008841:Actin Cytoskeleton / Q000502:physiology* / Q000648:ultrastructure; D000199:Actins / Q000502:physiology; D000818:Animals; D002460:Cell Line; D004847:Epithelial Cells; D005353:Fibronectins / Q000096:biosynthesis* / Q000235:genetics*; D019204:GTP-Binding Proteins / Q000378:metabolism*; D019012:Integrin beta1 / Q000737:chemistry / Q000235:genetics / Q000502:physiology*; D046911:Macromolecular Substances; D011994:Recombinant Proteins / Q000378:metabolism; D013961:Thyroid Gland; D014162:Transfection; D020742:rhoA GTP-Binding Protein
D000199:Actins; D005353:Fibronectins; D019012:Integrin beta1; D046911:Macromolecular Substances; D011994:Recombinant Proteins; D019204:GTP-Binding Proteins; D020742:rhoA GTP-Binding Protein