title: Requirement of LIM domains for the transient accumulation of paxillin at damaged stress fibres.
abstract: Cells recognize and respond to changes in intra- and extracellular mechanical conditions to maintain their mechanical homeostasis. Linear contractile bundles of actin filaments and myosin II known as stress fibres (SFs) mediate mechanical signals. Mechanical cues such as excessive stress driven by myosin II and/or external force may damage SFs and induce the local transient accumulation of SF-repair complexes (zyxin and VASP) at the damaged sites. Using an atomic force microscope mounted on a fluorescence microscope, we applied mechanical damage to cells expressing fluorescently tagged cytoskeletal proteins and recorded the subsequent mobilization of SF-repair complexes. We found that a LIM protein, paxillin, transiently accumulated at the damaged sites earlier than zyxin, while paxillin knockdown did not affect the kinetics of zyxin translocation. The C-terminal half of paxillin, comprising four-tandem LIM domains, can still translocate to damaged sites on SFs, suggesting that the LIM domain is essential for the mechanosensory function of paxillin. Our findings demonstrate a crucial role of the LIM domain in mechanosensing LIM proteins.
authors.affiliation: Innovation Laboratory, Tokyo Institute of Technology , S2-8, 4259 Nagatsuta-cho, Midori-ku, Yokohama 226-8503 , Japan ; Present address: Imaging Research Division, Bio-AFM Frontier Research Center, College of Science and Engineering, Kanazawa University, Kakuma-machi, Kanazawa 920-1192, Japan.
authors.forename: Takahiro
authors.initials: T
authors.lastname: Watanabe-Nakayama
authors.forename: Masakazu
authors.initials: M
authors.lastname: Saito
authors.forename: Shin'ichi
authors.initials: S
authors.lastname: Machida
authors.forename: Kikuo
authors.initials: K
authors.lastname: Kishimoto
authors.forename: Rehana
authors.initials: R
authors.lastname: Afrin
authors.forename: Atsushi
authors.initials: A
authors.lastname: Ikai
keywords: LIM domains; Mechanosensors; Repair of stress fibres