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Direct observation of DNA rotation during transcription by Escherichia coli RNA polymerase

Author

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  • Yoshie Harada

    (Faculty of Science and Technology, Keio University
    CREST (Core Research for Evolutional Science and Technology) “Genetic Programming” Team 13
    The Tokyo Metropolitan Institute of Medical Science)

  • Osamu Ohara

    (Kazusa DNA Research Institute)

  • Akira Takatsuki

    (Faculty of Science and Technology, Keio University)

  • Hiroyasu Itoh

    (CREST (Core Research for Evolutional Science and Technology) “Genetic Programming” Team 13
    Tsukuba Research Laboratory, Hamamatsu Photonics KK)

  • Nobuo Shimamoto

    (Structural Biology Center, National Institute of Genetics)

  • Kazuhiko Kinosita

    (Faculty of Science and Technology, Keio University
    CREST (Core Research for Evolutional Science and Technology) “Genetic Programming” Team 13)

Abstract

Helical filaments driven by linear molecular motors are anticipated to rotate around their axis, but rotation consistent with the helical pitch has not been observed. 14S dynein1 and non-claret disjunctional protein (ncd)2 rotated a microtubule more efficiently than expected for its helical pitch, and myosin rotated an actin filament only poorly3. For DNA-based motors such as RNA polymerase, transcription-induced supercoiling of DNA4 supports the general picture of tracking along the DNA helix5. Here we report direct and real-time optical microscopy measurements of rotation rate that are consistent with high-fidelity tracking. Single RNA polymerase molecules attached to a glass surface rotated DNA for >100 revolutions around the right-handed screw axis of the double helix with a rotary torque of >5 pN nm. This real-time observation of rotation opens the possibility of resolving individual transcription steps.

Suggested Citation

  • Yoshie Harada & Osamu Ohara & Akira Takatsuki & Hiroyasu Itoh & Nobuo Shimamoto & Kazuhiko Kinosita, 2001. "Direct observation of DNA rotation during transcription by Escherichia coli RNA polymerase," Nature, Nature, vol. 409(6816), pages 113-115, January.
    • Handle: RePEc:nat:nature:v:409:y:2001:i:6816:d:10.1038_35051126
    DOI: 10.1038/35051126
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    Cited by:

    1. Ferdinand Greiss & Nicolas Lardon & Leonie Schütz & Yoav Barak & Shirley S. Daube & Elmar Weinhold & Vincent Noireaux & Roy Bar-Ziv, 2024. "A genetic circuit on a single DNA molecule as an autonomous dissipative nanodevice," Nature Communications, Nature, vol. 15(1), pages 1-12, December.
    2. Tamra Heberling & Lisa Davis & Jakub Gedeon & Charles Morgan & Tomáš Gedeon, 2016. "A Mechanistic Model for Cooperative Behavior of Co-transcribing RNA Polymerases," PLOS Computational Biology, Public Library of Science, vol. 12(8), pages 1-38, August.

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