title: Differential effect of actomyosin relaxation on the dynamic properties of focal adhesion proteins.
abstract: Treatment of cultured cells with inhibitors of actomyosin contractility induces rapid deterioration of stress fibers, and disassembly of the associated focal adhesions (FAs). In this study, we show that treatment with the Rho kinase inhibitor Y-27632, which blocks actomyosin contractility, induces disarray in the FA-associated actin bundles, followed by the differential dissociation of eight FA components from the adhesion sites. Live-cell microscopy indicated that the drug triggers rapid dissociation of VASP and zyxin from FAs (τ values of 7-8 min), followed by talin, paxillin and ILK (τ ~16 min), and then by FAK, vinculin and kindlin-2 (τ = 25-28 min). Examination of the molecular kinetics of the various FA constituents, using Fluorescence Recovery After Photobleaching (FRAP), in the absence of or following short-term treatment with the drug, revealed major changes in the kon and koff values of the different proteins tested, which are in close agreement with their differential dissociation rates from the adhesion sites. These findings indicate that mechanical, actomyosin-generated forces differentially regulate the molecular kinetics of individual FA-associated molecules, and thereby modulate FA composition and stability.
authors.affiliation: Department of Molecular Cell Biology, Weizmann Institute of Science, Rehovot, Israel.
authors.forename: Irena
authors.initials: I
authors.lastname: Lavelin
authors.forename: Haguy
authors.initials: H
authors.lastname: Wolfenson
authors.forename: Israel
authors.initials: I
authors.lastname: Patla
authors.forename: Yoav I
authors.initials: YI
authors.lastname: Henis
authors.forename: Ohad
authors.initials: O
authors.lastname: Medalia
authors.forename: Tova
authors.initials: T
authors.lastname: Volberg
authors.forename: Ariel
authors.initials: A
authors.lastname: Livne
authors.forename: Zvi
authors.initials: Z
authors.lastname: Kam
authors.forename: Benjamin
authors.initials: B
authors.lastname: Geiger
mesh_terms: D008841:Actin Cytoskeleton / Q000187:drug effects / Q000378:metabolism*; D000205:Actomyosin / Q000037:antagonists & inhibitors / Q000378:metabolism*; D000577:Amides / Q000494:pharmacology*; D000818:Animals; D002460:Cell Line; D022001:Focal Adhesions / Q000187:drug effects / Q000378:metabolism*; D006801:Humans; D047428:Protein Kinase Inhibitors / Q000494:pharmacology*; D011725:Pyridines / Q000494:pharmacology*; D051381:Rats; D054460:rho-Associated Kinases / Q000037:antagonists & inhibitors*
chemical_list: D000577:Amides; D047428:Protein Kinase Inhibitors; D011725:Pyridines; C108830:Y 27632; D000205:Actomyosin; D054460:rho-Associated Kinases
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