Fast microscopical dissection of action scenes played by Escherichia coli RNA polymerase

被引:16
作者
Suzuki, Yuki [1 ]
Shin, Minsang [1 ]
Yoshida, Aiko [1 ]
Yoshimura, Shige H. [1 ]
Takeyasu, Kunio [1 ]
机构
[1] Kyoto Univ, Grad Sch Biostudies, Lab Plasma Membrane & Nucl Signaling, Sakyo Ku, Kyoto 6068501, Japan
基金
日本学术振兴会;
关键词
Atomic force microscopy; Fast-scanning atomic force microscopy; RNA polymerase; Protein-DNA interaction; ATOMIC-FORCE MICROSCOPY; CONFORMATIONAL-CHANGES; DNA INTERACTIONS; START SITE; REAL-TIME; TRANSCRIPTION; COMPLEXES; VISUALIZATION; DIFFUSION; REVEALS;
D O I
10.1016/j.febslet.2012.06.033
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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. (C) 2012 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.
引用
收藏
页码:3187 / 3192
页数:6
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