title: A programmable DNA origami nanospring that reveals force-induced adjacent binding of myosin VI heads.
abstract: Mechanosensitive biological nanomachines such as motor proteins and ion channels regulate diverse cellular behaviour. Combined optical trapping with single-molecule fluorescence imaging provides a powerful methodology to clearly characterize the mechanoresponse, structural dynamics and stability of such nanomachines. However, this system requires complicated experimental geometry, preparation and optics, and is limited by low data-acquisition efficiency. Here we develop a programmable DNA origami nanospring that overcomes these issues. We apply our nanospring to human myosin VI, a mechanosensory motor protein, and demonstrate nanometre-precision single-molecule fluorescence imaging of the individual motor domains (heads) under force. We observe force-induced transitions of myosin VI heads from non-adjacent to adjacent binding, which correspond to adapted roles for low-load and high-load transport, respectively. Our technique extends single-molecule studies under force and clarifies the effect of force on biological processes.
authors.affiliation: Quantitative Biology Center, RIKEN, OLABB, 6-2-3, Furuedai, Suita, Osaka 5650874, Japan.|Graduate School of Frontier Biosciences, Osaka University, Suita, Osaka 5650871, Japan.
authors.forename: M
authors.identifier: 0000-0002-8893-1103
authors.initials: M
authors.lastname: Iwaki
authors.affiliation: Department of Cancer Biology, Dana-Farber Cancer Institute, Boston, Massachusetts 02115, USA.|Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, Massachusetts 02115, USA.|Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, Massachusetts 02115, USA.
authors.forename: S F
authors.initials: SF
authors.lastname: Wickham
authors.affiliation: School of Frontier Sciences, The University of Tokyo, Kashiwa, Chiba 2778561, Japan.
authors.forename: K
authors.initials: K
authors.lastname: Ikezaki
authors.affiliation: Quantitative Biology Center, RIKEN, OLABB, 6-2-3, Furuedai, Suita, Osaka 5650874, Japan.|Graduate School of Frontier Biosciences, Osaka University, Suita, Osaka 5650871, Japan.|Center for Information and Neural Networks, NICT, Suita, Osaka 5650874, Japan.
authors.forename: T
authors.initials: T
authors.lastname: Yanagida
authors.affiliation: Department of Cancer Biology, Dana-Farber Cancer Institute, Boston, Massachusetts 02115, USA.|Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, Massachusetts 02115, USA.|Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, Massachusetts 02115, USA.
authors.forename: W M
authors.initials: WM
authors.lastname: Shih
mesh_terms: D001692:Biological Transport; D006801:Humans; D040542:Mechanotransduction, Cellular; D018995:Myosin Heavy Chains / Q000737:chemistry* / Q000648:ultrastructure; D036103:Nanotechnology; D061848:Optical Imaging
chemical_list: C089785:myosin VI; D018995:Myosin Heavy Chains