ATP对RuvA-RuvB-霍利迪连接体DNA复合物形成的协同作用。

Synergistic effect of ATP for RuvA-RuvB-Holliday junction DNA complex formation.

作者信息

Iwasa Takuma, Han Yong-Woon, Hiramatsu Ryo, Yokota Hiroaki, Nakao Kimiko, Yokokawa Ryuji, Ono Teruo, Harada Yoshie

机构信息

Institute for Integrated Cell-Materials Science (WPI-iCeMS), Kyoto University, Sakyo, Kyoto 606-8501, Japan.

Graduate School of Biostudies, Kyoto University, Sakyo, Kyoto 606-8501, Japan.

出版信息

Sci Rep. 2015 Dec 14;5:18177. doi: 10.1038/srep18177.

The Escherichia coli RuvB hexameric ring motor proteins, together with RuvAs, promote branch migration of Holliday junction DNA. Zero mode waveguides (ZMWs) constitute of nanosized holes and enable the visualization of a single fluorescent molecule under micromolar order of the molecules, which is applicable to characterize the formation of RuvA-RuvB-Holliday junction DNA complex. In this study, we used ZMWs and counted the number of RuvBs binding to RuvA-Holliday junction DNA complex. Our data demonstrated that different nucleotide analogs increased the amount of Cy5-RuvBs binding to RuvA-Holliday junction DNA complex in the following order: no nucleotide, ADP, ATPγS, and mixture of ADP and ATPγS. These results suggest that not only ATP binding to RuvB but also ATP hydrolysis by RuvB facilitates a stable RuvA-RuvB-Holliday junction DNA complex formation.

大肠杆菌RuvB六聚体环状马达蛋白与RuvA一起促进霍利迪连接体DNA的分支迁移。零模式波导（ZMW）由纳米尺寸的孔构成，能够在微摩尔级别的分子浓度下可视化单个荧光分子，这适用于表征RuvA-RuvB-霍利迪连接体DNA复合物的形成。在本研究中，我们使用ZMW并对结合到RuvA-霍利迪连接体DNA复合物上的RuvB数量进行了计数。我们的数据表明，不同的核苷酸类似物按照以下顺序增加了Cy5-RuvB与RuvA-霍利迪连接体DNA复合物的结合量：无核苷酸、ADP、ATPγS以及ADP和ATPγS的混合物。这些结果表明，不仅ATP与RuvB的结合，而且RuvB对ATP的水解都有助于稳定的RuvA-RuvB-霍利迪连接体DNA复合物的形成。