title: Documentation and localization of force-mediated filamin A domain perturbations in moving cells.
abstract: Endogenously and externally generated mechanical forces influence diverse cellular activities, a phenomenon defined as mechanotransduction. Deformation of protein domains by application of stress, previously documented to alter macromolecular interactions in vitro, could mediate these effects. We engineered a photon-emitting system responsive to unfolding of two repeat domains of the actin filament (F-actin) crosslinker protein filamin A (FLNA) that binds multiple partners involved in cell signalling reactions and validated the system using F-actin networks subjected to myosin-based contraction. Expressed in cultured cells, the sensor-containing FLNA construct reproducibly reported FLNA domain unfolding strikingly localized to dynamic, actively protruding, leading cell edges. The unfolding signal depends upon coherence of F-actin-FLNA networks and is enhanced by stimulating cell contractility. The results establish protein domain distortion as a bona fide mechanism for mechanotransduction in vivo.
authors.affiliation: Translational Medicine Division, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts 02445, USA.
authors.forename: Fumihiko
authors.initials: F
authors.lastname: Nakamura
authors.affiliation: Translational Medicine Division, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts 02445, USA.
authors.forename: Mia
authors.initials: M
authors.lastname: Song
authors.affiliation: Translational Medicine Division, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts 02445, USA.
authors.forename: John H
authors.initials: JH
authors.lastname: Hartwig
authors.affiliation: Translational Medicine Division, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts 02445, USA.
authors.forename: Thomas P
authors.initials: TP
authors.lastname: Stossel
mesh_terms: D000199:Actins / Q000502:physiology; D000818:Animals; D001696:Biomechanical Phenomena / Q000502:physiology; D019556:COS Cells; D002465:Cell Movement / Q000502:physiology*; D002478:Cells, Cultured; D002522:Chlorocebus aethiops; D064448:Filamins / Q000737:chemistry* / Q000502:physiology*; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods; D066298:In Vitro Techniques; D007668:Kidney / Q000166:cytology / Q000502:physiology*; D040542:Mechanotransduction, Cellular / Q000502:physiology*; D055095:Optics and Photonics / Q000379:methods
chemical_list: D000199:Actins; D064448:Filamins