title: Fibronectin's central cell-binding domain supports focal adhesion formation and Rho signal transduction.
abstract: Fibroblast adhesion to fibronectin (FN) induces formation of focal adhesions (FAs), structures that have significant effect on cell migration and signaling. FA formation requires actomyosin-based contractility that is regulated by Rho-dependent myosin light chain (MLC) phosphorylation. Previous studies indicated that the FN central cell-binding (and integrin-binding) domain (CBD) is insufficient for FA formation and that the major heparin-binding domain (HepII) facilitates FA formation in a Rho-dependent manner. We describe here conditions under which FN CBD alone is sufficient for FA formation in both human dermal fibroblasts and the FN-null murine fibroblasts. CBD-mediated FA formation is dependent on its surface adsorption and the adhesion activity of the cells. Attachment of FN-null fibroblasts to CBD elicits the same biphasic regulation of Rho activity as seen on intact FN, whereas adhesion to HepII alone does not activate Rho. Activation of Rho requires high levels of integrin occupancy. However, FN or CBD may induce FAs without increased activation of Rho (i.e. the basal level of GTP-Rho induces sufficient phospho-MLC for FA assembly under this condition). In contrast, adhesion to HepII alone does not sustain MLC phosphorylation. Pulse stimulation of cells on CBD or HepII with lysophosphatidic acid elevates Rho GTP loading to the same level, but the lysophosphatidic acid-stimulated MLC phosphorylation is significantly lower in cells on HepII than on CBD. Coating HepII with suboptimal concentrations of CBD induces FAs without increased activation of Rho. Therefore, FN CBD can support FA formation and generate contraction by activating Rho or by facilitating Rho downstream signaling.
authors.affiliation: Department of Dermatology, State University of New York at Stony Brook, Stony Brook, New York 11794, USA.
authors.forename: Ruixue
authors.initials: R
authors.lastname: Wang
authors.forename: Richard A F
authors.initials: RA
authors.lastname: Clark
authors.forename: Deane F
authors.initials: DF
authors.lastname: Mosher
authors.forename: Xiang-Dong
authors.initials: XD
authors.lastname: Ren
mesh_terms: D000818:Animals; D001665:Binding Sites; D002448:Cell Adhesion / Q000502:physiology; D003001:Cloning, Molecular; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism; D005353:Fibronectins / Q000172:deficiency / Q000502:physiology*; D006801:Humans; D051379:Mice; D018613:Microscopy, Confocal; D018994:Myosin Light Chains / Q000378:metabolism; D010446:Peptide Fragments / Q000494:pharmacology; D011994:Recombinant Proteins / Q000378:metabolism; D012867:Skin / Q000166:cytology / Q000378:metabolism*
chemical_list: D005353:Fibronectins; D018994:Myosin Light Chains; D010446:Peptide Fragments; D011994:Recombinant Proteins