Stanley Louise K, Szczelkun Mark D
DNA-Protein Interactions Unit, Department of Biochemistry, University of Bristol, Bristol, BS8 1TD, UK.
Nucleic Acids Res. 2006;34(16):4387-94. doi: 10.1093/nar/gkl569. Epub 2006 Aug 26.
With the aim of investigating how motor proteins negotiate DNA nanostructures, we produced test circuits based on recombination intermediates in which 1D translocation across a Holliday junction (HJ) could be assessed by subsequent triplex displacement signals on each DNA arm. Using the EcoR124I restriction-modification enzyme, a 3'-5' double-strand DNA (dsDNA) translocase, we could show that the motor will tend to follow its translocated strand across a junction. Nonetheless, as the frequency of junction bypass events increases, the motor will occasionally jump tracks.
为了研究运动蛋白如何穿越DNA纳米结构,我们基于重组中间体构建了测试电路,其中通过每个DNA臂上随后的三链体置换信号可以评估在霍利迪连接体(HJ)上的一维易位。使用EcoR124I限制修饰酶(一种3'-5'双链DNA(dsDNA)转位酶),我们可以证明该运动蛋白倾向于沿着其易位链穿越连接体。尽管如此,随着连接体旁路事件频率的增加,该运动蛋白偶尔会跳轨。