Direct Visualization of the Movement of a Single T7 RNA Polymerase and Transcription on a DNA Nanostructure

Authors

  • Dr. Masayuki Endo,

    Corresponding author
    1. Institute for Integrated Cell-Material Sciences (WPI-iCeMS), Kyoto University, Yoshida-ushinomiyacho, Sakyo-ku, Kyoto 606-8501 (Japan)
    2. CREST, Japan Science and Technology Corporation (JST), Sanbancho, Chiyoda-ku, Tokyo 102-0075 (Japan)
    • Institute for Integrated Cell-Material Sciences (WPI-iCeMS), Kyoto University, Yoshida-ushinomiyacho, Sakyo-ku, Kyoto 606-8501 (Japan)
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  • Koichi Tatsumi,

    1. Department of Chemistry, Graduate School of Science, Kyoto University, Kitashirakawa-oiwakecho, Sakyo-ku, Kyoto 606-8502 (Japan)
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  • Kosuke Terushima,

    1. Department of Chemistry, Graduate School of Science, Kyoto University, Kitashirakawa-oiwakecho, Sakyo-ku, Kyoto 606-8502 (Japan)
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  • Yousuke Katsuda,

    1. Department of Chemistry, Graduate School of Science, Kyoto University, Kitashirakawa-oiwakecho, Sakyo-ku, Kyoto 606-8502 (Japan)
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  • Kumi Hidaka,

    1. Department of Chemistry, Graduate School of Science, Kyoto University, Kitashirakawa-oiwakecho, Sakyo-ku, Kyoto 606-8502 (Japan)
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  • Prof. Yoshie Harada,

    1. Institute for Integrated Cell-Material Sciences (WPI-iCeMS), Kyoto University, Yoshida-ushinomiyacho, Sakyo-ku, Kyoto 606-8501 (Japan)
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  • Prof. Hiroshi Sugiyama

    Corresponding author
    1. Institute for Integrated Cell-Material Sciences (WPI-iCeMS), Kyoto University, Yoshida-ushinomiyacho, Sakyo-ku, Kyoto 606-8501 (Japan)
    2. Department of Chemistry, Graduate School of Science, Kyoto University, Kitashirakawa-oiwakecho, Sakyo-ku, Kyoto 606-8502 (Japan)
    3. CREST, Japan Science and Technology Corporation (JST), Sanbancho, Chiyoda-ku, Tokyo 102-0075 (Japan)
    • Institute for Integrated Cell-Material Sciences (WPI-iCeMS), Kyoto University, Yoshida-ushinomiyacho, Sakyo-ku, Kyoto 606-8501 (Japan)
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  • This work was supported by Core Research for Evolutional Science and Technology (CREST) of JST and Grant-in-Aid for Scientific Research from MEXT (Japan). Financial support from Sumitomo foundation and Iketani Science and Technology Foundation to M.E. is also acknowledged.

Abstract

original image

Schnappschüsse der Transkription: Die Bewegung eines T7-RNA-Polymerase(RNAP)-Moleküls entlang eines DNA-Doppelstrang(dsDNA)-Templats auf einer DNA-Origami-Plattform (grün, siehe Schema) wurde mit Hochgeschwindigkeits-AFM verfolgt. An einem dsDNA-Templat mit einer Länge von einer Kilobase, das den T7-Promotor enthielt, wurden AFM-Bilder von der Transkription durch die RNAP aufgenommen, und die produzierte biotinylierte RNA wurde durch Streptavidin-Markierung nachgewiesen.

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