Trimers of the fibronectin cell adhesion domain localize to actin filament bundles and undergo rearward translocation.
Previous studies have shown that small beads coated with FN7-10, a four-domain cell adhesion fragment of fibronectin, bind to cell surfaces and translocate rearward. Here we investigate whether soluble constructs containing two to five FN7-10 units might be sufficient for activity. We have produced a monomer, three forms of dimers, a trimer and a pentamer of FN7-10, on the end of spacer arms. These oligomers could bind small clusters of up to five integrins. Fluorescence microscopy showed that the trimer and pentamer bound strongly to the cell surface, and within 5 minutes were prominently localized to actin fiber bundles. Monomers and dimers showed only diffuse localization. Beads coated with a low concentration (probably one complex per bead) of trimer or pentamer showed prolonged binding and rearward translocation, presumably with the translocating actin cytskeleton. Beads containing monomer or dimer showed only brief binding and diffusive movements. We conclude that clusters of three integrin-binding ligands are necessary and sufficient for coupling to and translocating with the actin cytoskeleton.
Coussen
Françoise
F
Department of Cell Biology, Box 3709, Duke University Medical Center, Durham, NC 27710, USA.
Choquet
Daniel
D
Sheetz
Michael P
MP
Erickson
Harold P
HP
D016475:3T3 Cells; D008841:Actin Cytoskeleton / Q000378:metabolism*; D000818:Animals; D002448:Cell Adhesion / Q000502:physiology*; D002462:Cell Membrane / Q000378:metabolism*; D005057:Eukaryotic Cells / Q000166:cytology / Q000378:metabolism*; D005109:Extracellular Matrix / Q000378:metabolism*; D005353:Fibronectins / Q000378:metabolism* / Q000648:ultrastructure; D039081:Integrin alpha5beta1 / Q000378:metabolism; D016023:Integrins / Q000378:metabolism; D051379:Mice; D008863:Microspheres; D010446:Peptide Fragments / Q000737:chemistry / Q000648:ultrastructure; D011108:Polymers / Q000737:chemistry; D011485:Protein Binding / Q000502:physiology; D017434:Protein Structure, Tertiary / Q000502:physiology; D021381:Protein Transport / Q000502:physiology
D005353:Fibronectins; D039081:Integrin alpha5beta1; D016023:Integrins; D010446:Peptide Fragments; D011108:Polymers