title: Vinculin tension distributions of individual stress fibers within cell-matrix adhesions.
abstract: Actomyosin stress fibers (SFs) enable cells to exert traction on planar extracellular matrices (ECMs) by tensing focal adhesions (FAs) at the cell-ECM interface. Although it is widely appreciated that the spatial and temporal distribution of these tensile forces play key roles in polarity, motility, fate choice, and other defining cell behaviors, virtually nothing is known about how an individual SF quantitatively contributes to tensile loads borne by specific molecules within associated FAs. We address this key open question by using femtosecond laser ablation to sever single SFs in cells while tracking tension across vinculin using a molecular optical sensor. We show that disruption of a single SF reduces tension across vinculin in FAs located throughout the cell, with enriched vinculin tension reduction in FAs oriented parallel to the targeted SF. Remarkably, however, some subpopulations of FAs exhibit enhanced vinculin tension upon SF irradiation and undergo dramatic, unexpected transitions between tension-enhanced and tension-reduced states. These changes depend strongly on the location of the severed SF, consistent with our earlier finding that different SF pools are regulated by distinct myosin activators. We critically discuss the extent to which these measurements can be interpreted in terms of whole-FA tension and traction and propose a model that relates SF tension to adhesive loads and cell shape stability. These studies represent the most direct and high-resolution intracellular measurements of SF contributions to tension on specific FA proteins to date and offer a new paradigm for investigating regulation of adhesive complexes by cytoskeletal force.
authors.affiliation: Department of Bioengineering, University of California Berkeley, Berkeley, CA 94720, USA.
authors.forename: Ching-Wei
authors.initials: CW
authors.lastname: Chang
authors.forename: Sanjay
authors.initials: S
authors.lastname: Kumar
mesh_terms: D001696:Biomechanical Phenomena; D002448:Cell Adhesion; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000528:radiation effects; D016764:Cell Polarity / Q000528:radiation effects; D048430:Cell Shape / Q000528:radiation effects; D003599:Cytoskeleton / Q000378:metabolism*; D031541:Fluorescence Resonance Energy Transfer; D022001:Focal Adhesions / Q000378:metabolism* / Q000528:radiation effects / Q000648:ultrastructure; D006801:Humans; D053685:Laser Therapy; D008954:Models, Biological; D021381:Protein Transport / Q000528:radiation effects; D022502:Stress Fibers / Q000737:chemistry / Q000378:metabolism* / Q000528:radiation effects; D016596:Vinculin / Q000737:chemistry / Q000378:metabolism*
chemical_list: D016596:Vinculin
keywords: Cell mechanics; Fluorescence resonance energy transfer; Focal adhesions; Laser ablation; Mechanotransduction; Stress fibers