title: Position-dependent linkages of fibronectin- integrin-cytoskeleton.
abstract: Position-dependent cycling of integrin interactions with both the cytoskeleton and extracellular matrix (ECM) is essential for cell spreading, migration, and wound healing. Whether there are regional changes in integrin concentration, ligand affinity or cytoskeleton crosslinking of liganded integrins has been unclear. Here, we directly demonstrate a position-dependent binding and release cycle of fibronectin-integrin-cytoskeleton interactions with preferential binding at the front of motile 3T3 fibroblasts and release at the endoplasm-ectoplasm boundary. Polystyrene beads coated with low concentrations of an integrin-binding fragment of fibronectin (fibronectin type III domains 7-10) were 3-4 times more likely to bind to integrins when placed within 0.5 microns vs. 0.5-3 microns from the leading edge. Integrins were not concentrated at the leading edge, nor did anti-integrin antibody-coated beads bind preferentially at the leading edge. However, diffusing liganded integrins attached to the cytoskeleton preferentially at the leading edge. Cytochalasin inhibited edge binding, which suggested that cytoskeleton binding to the integrins could alter the avidity for ligand beads. Further, at the ectoplasm-endoplasm boundary, the velocity of bead movement decreased, diffusive motion increased, and approximately one-third of the beads were released into the medium. We suggest that cytoskeleton linkage of liganded integrins stabilizes integrin-ECM bonds at the front whereas release of cytoskeleton-integrin links weakens integrin-ECM bonds at the back of lamellipodia.
authors.affiliation: Department of Cell Biology, Duke University Medical Center, Durham, NC 27710, USA.
authors.forename: T
authors.initials: T
authors.lastname: Nishizaka
authors.forename: Q
authors.initials: Q
authors.lastname: Shi
authors.forename: M P
authors.initials: MP
authors.lastname: Sheetz
mesh_terms: D016475:3T3 Cells / Q000166:cytology / Q000378:metabolism; D000818:Animals; D002462:Cell Membrane / Q000378:metabolism; D002465:Cell Movement / Q000187:drug effects; D003571:Cytochalasin B / Q000494:pharmacology; D003599:Cytoskeleton / Q000378:metabolism*; D005353:Fibronectins / Q000378:metabolism*; D019012:Integrin beta1 / Q000378:metabolism; D016023:Integrins / Q000378:metabolism*; D051379:Mice; D008863:Microspheres; D011485:Protein Binding / Q000187:drug effects
chemical_list: D005353:Fibronectins; D019012:Integrin beta1; D016023:Integrins; D003571:Cytochalasin B