HCV解旋酶长路径运动行为的单分子研究

HCV解旋酶在HCV病毒复制中起着关键作用，是抗HCV药物研发的重要靶蛋白。深入研究HCV解旋酶的生物功能与作用机制，对以解旋酶为靶点的抗HCV药物研发具有重要意义。已有研究通过光阱、磁镊等单分子技术获得了解旋酶运动轨迹、能量配给、路径依赖等重要信息，但目前尚无解旋酶长路径的单分子观测模型的报道。申报人有较丰富的单分子观测经验，并在前期已完成HCV解旋酶的表达纯化，本项目拟借助申报人已掌握的DNA origami技术，在长矩形DNA origami上搭载HCV来源的长链RNA，作为HCV解旋酶的运动轨道，构建长路径单分子观测模型，并运用此模型对解旋酶长路径的运动特征进行实时观测，进而探讨HCV解旋酶作用的协同效应和对RNA结合蛋白的剥离过程。项目的完成将进一步阐明HCV解旋酶的生物功能与作用机制，为其他解旋酶的研究提供重要参考，并为以解旋酶为靶点的抗HCV药物研发提供重要的理论依据。

HCV helicase, palying a critical role in HCV viral replication, is an important target protein in the development of anti-HCV drugs. Therefore, it is vital to further investigate the biological function and working mechanism of HCV helicase for developing the anti-HCV drugs. The information of helicase’s motion trail, energy supply and path dependence by single-molecule techniques such as optical tweezers and magnetic tweezers, have been well evaluated. However, It's rarely reported about the single-molecule observation model for observing the long-path movement of HCV helicase. In the previous work, we have accumulated rich experience on the single molecular observation, meanwhile, some HCV helicase have been expressed and purified. Hence, in this project, basing on our DNA origami oberving skill, we plan to build a DNA origami observation model for observing the long-path behavior of HCV helicase. Meanwhile, by this observation model, we will try to get more information on the cooperativity and protein displacement of HCV helicase. The completion of the project could deepen the understanding of HCV helicase’s biological functions and working mechanism. Moreover, it will provide an important reference for the research on other helicases and theoretical foundation for the development of anti-HCV drugs.