Laboratory of Plasma Membrane and Nuclear Signaling, Graduate School of Biostudies, Kyoto University, Sakyo-ku, Kyoto, Japan.
FEBS Lett. 2012 Sep 21;586(19):3187-92. doi: 10.1016/j.febslet.2012.06.033. Epub 2012 Jul 4.
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.
使用快速扫描原子力显微镜，我们以每秒 1-2 帧的扫描速度直接观察到大肠杆菌 RNA 聚合酶 (RNAP) 与 DNA 的相互作用。分析表明，RNAP 不仅可以通过滑动，还可以通过跳跃和/或片段转移来定位启动子区域。当向停滞复合物中加入 0.05mM 的 NTPs 时，RNAP 分子以平均 15 个核苷酸/秒的速度单向拉动模板 DNA。本方法有可能适用于检查各种蛋白质-核酸相互作用，特别是那些涉及基因调控过程的相互作用。
