Fast microscopical dissection of action scenes played by Escherichia coli RNA polymerase |
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Authors: | Yuki Suzuki Minsang Shin Aiko Yoshida Shige H Yoshimura Kunio Takeyasu |
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Institution: | Laboratory of Plasma Membrane and Nuclear Signaling, Graduate School of Biostudies, Kyoto University, Yoshida-Konoe-cho, Sakyo-ku, Kyoto 606-8501, Japan |
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Abstract: | Using fast-scanning atomic force microscopy, we directly visualized the interaction of Escherichia coli RNA polymerase (RNAP) with DNA at the scan rate of 1-2 frames per second. The analyses showed that the RNAP can locate the promoter region not only by sliding but also by hopping and/or segmental transfer. Upon the addition of 0.05 mM NTPs to the stalled complex, the RNAP molecule pulled the template DNA uni-directionally at the rates of 15 nucleotides/s on average. The present method is potentially applicable to examine a variety of protein-nucleic acid interactions, especially those involved in the process of gene regulation. |
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Keywords: | Atomic force microscopy Fast-scanning atomic force microscopy RNA polymerase Protein-DNA interaction |
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