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

Snapshots of transcription: Movement of a single molecule of T7 RNA polymerase (RNAP) along a double-stranded DNA (dsDNA) template attached to a DNA origami platform (green, see scheme) was recorded using high-speed AFM. A one kilobase template dsDNA containing the T7 promoter was used to record AFM images of transcription of the template dsDNA by RNAP and the resulting biotinylated RNA product was detected by streptavidin-labeling.

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