Dynamics of ligand-induced, Rac1-dependent anchoring of cadherins to the actin cytoskeleton.
Cadherin receptors are key morphoregulatory molecules during development. To dissect their mode of action, we developed an approach based on the use of myogenic C2 cells and beads coated with an Ncad-Fc ligand, allowing us to mimic cadherin-mediated adhesion. We used optical tweezers and video microscopy to investigate the dynamics of N-cadherin anchoring within the very first seconds of bead-cell contact. The analysis of the bead movement by single-particle tracking indicated that N-cadherin molecules were freely diffusive in the first few seconds after bead binding. The beads rapidly became diffusion-restricted and underwent an oriented rearward movement as a result of N-cadherin anchoring to the actin cytoskeleton. The kinetics of anchoring were dependent on ligand density, suggesting that it was an inducible process triggered by active cadherin recruitment. This anchoring was inhibited by the dominant negative form of Rac1, but not that of Cdc42. The Rac1 mutant had no effect on cell contact formation or cadherin-catenin complex recruitment, but did inhibit actin recruitment. Our results suggest that cadherin anchoring to the actin cytoskeleton is an adhesion-triggered, Rac1-regulated process enabling the transduction of mechanical forces across the cell membrane; they uncover novel aspects of the action of cadherins in cell sorting, cell migration, and growth cone navigation.
Lambert
Mireille
M
Signalisation et Différenciation Cellulaires dans les Systèmes Nerveux et Musculaire, INSERM U440, Université Paris 6, Institut du Fer à Moulin, 75005 Paris, France.
Choquet
Daniel
D
Mège
René-Marc
RM
D000199:Actins / Q000378:metabolism*; D000818:Animals; D015820:Cadherins / Q000235:genetics / Q000378:metabolism*; D002448:Cell Adhesion / Q000187:drug effects; D002460:Cell Line; D002645:Chickens; D003599:Cytoskeleton / Q000378:metabolism*; D007141:Immunoglobulin Fc Fragments / Q000235:genetics / Q000378:metabolism; D007700:Kinetics; D008024:Ligands; D008148:Lovastatin / Q000494:pharmacology; D051379:Mice; D008863:Microspheres; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D020764:cdc42 GTP-Binding Protein / Q000378:metabolism; D020830:rac1 GTP-Binding Protein / Q000502:physiology*
D000199:Actins; D015820:Cadherins; D007141:Immunoglobulin Fc Fragments; D008024:Ligands; D011993:Recombinant Fusion Proteins; D008148:Lovastatin; D020764:cdc42 GTP-Binding Protein; D020830:rac1 GTP-Binding Protein