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

Nature volume 409, pages 113–115 (2001)Cite this article

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.

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Figure 1: Observation of DNA rotation by RNA polymerase.
Figure 2: Time courses of bead rotation at various [NTP].
Figure 3: Comparison of rotation and transcription rates.

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Acknowledgements

We thank M. Susa for help in transcription analysis; A. Ishihama, S. Ishiwata, G. W. Feigenson and members of Team 13 for comments; and H. Umezawa for laboratory management. This work was supported in part by Grants-in-Aid from Ministry of Education, Science, Sports and Culture of Japan, Hayashi Memorial Foundation for Female Natural Scientists, and an Academic Frontier Promotional Project.

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Author notes

  1. Yoshie Harada

    Present address: Department of Molecular Physiology, The Tokyo Metropolitan Institute of Medical Science, 18-22, Honkomagome 3-chome, Bunkyo-ku, Tokyo, 113-8613, Japan

Authors and Affiliations

  1. Department of Physics, Faculty of Science and Technology, Keio University, Hiyoshi 3-14-1, Kohoku-ku, 223-8522, Yokohama, Japan

    Yoshie Harada, Akira Takatsuki & Kazuhiko Kinosita Jr

  2. CREST (Core Research for Evolutional Science and Technology) “Genetic Programming” Team 13, Nogawa 907, Miyamae-ku, 216-0001 , Kawasaki, Japan

    Yoshie Harada, Hiroyasu Itoh & Kazuhiko Kinosita Jr

  3. Kazusa DNA Research Institute, Yana 1532-3, Kisarazu, 292-0812, Japan

    Osamu Ohara

  4. Tsukuba Research Laboratory, Hamamatsu Photonics KK, Tokodai, 300-2635, Tsukuba, Japan

    Hiroyasu Itoh

  5. Structural Biology Center, National Institute of Genetics, Mishima, 411-8540, Japan

    Nobuo Shimamoto

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  1. Yoshie Harada
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Corresponding author

Correspondence to Yoshie Harada.

Supplementary information

41586_2001_BF35051126_MOESM1_ESM.doc

Analyses of transcription rates in solution (estimation by gel electrophoresis) and on individual RNAP molecules attached to a glass surface (estimation from the reduction in the range of brownian motion of the end bead).

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Harada, Y., Ohara, O., Takatsuki, A. et al. Direct observation of DNA rotation during transcription by Escherichia coli RNA polymerase. Nature 409, 113–115 (2001). https://doi.org/10.1038/35051126

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