Direct observation of DNA rotation during transcription by Escherichia coli RNA polymerase.
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 dynein and non-claret disjunctional protein (ncd) rotated a microtubule more efficiently than expected for its helical pitch, and myosin rotated an actin filament only poorly. For DNA-based motors such as RNA polymerase, transcription-induced supercoiling of DNA supports the general picture of tracking along the DNA helix. 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.
Harada
Y
Y
Department of Physics, Faculty of Science and Technology, Keio University, Hiyoshi 3-14-1, Kohoku-ku, Yokohama 223-8522, Japan. yharada@rinshoken.or.jp
Ohara
O
O
Takatsuki
A
A
Itoh
H
H
Shimamoto
N
N
Kinosita
K
K
D004247:DNA / Q000737:chemistry / Q000378:metabolism*; D012321:DNA-Directed RNA Polymerases / Q000737:chemistry / Q000378:metabolism*; D004926:Escherichia coli / Q000201:enzymology; D005456:Fluorescent Dyes; D008280:Magnetics; D008853:Microscopy; D008863:Microspheres; D020409:Molecular Motor Proteins; D009690:Nucleic Acid Conformation; D012313:RNA / Q000378:metabolism; D012399:Rotation; D014158:Transcription, Genetic
D005456:Fluorescent Dyes; D020409:Molecular Motor Proteins; D012313:RNA; D004247:DNA; D012321:DNA-Directed RNA Polymerases