You Huijuan, Le Shimin, Chen Hu, Qin Linyan, Yan Jie
School of Pharmacy, Tongji Medical College, Huazhong University of Science and Technology;
Mechanobiology Institute, National University of Singapore; Department of Physics, National University of Singapore.
J Vis Exp. 2017 Sep 19(127):56328. doi: 10.3791/56328.
Non-canonical nucleic acid secondary structure G-quadruplexes (G4) are involved in diverse cellular processes, such as DNA replication, transcription, RNA processing, and telomere elongation. During these processes, various proteins bind and resolve G4 structures to perform their function. As the function of G4 often depends on the stability of its folded structure, it is important to investigate how G4 binding proteins regulate the stability of G4. This work presents a method to manipulate single G4 molecules using magnetic tweezers, which enables studies of the regulation of G4 binding proteins on a single G4 molecule in real time. In general, this method is suitable for a wide scope of applications in studies for proteins/ligands interactions and regulations on various DNA or RNA secondary structures.
非经典核酸二级结构G-四链体（G4）参与多种细胞过程，如DNA复制、转录、RNA加工和端粒延长。在这些过程中，各种蛋白质结合并解开G4结构以执行其功能。由于G4的功能通常取决于其折叠结构的稳定性，因此研究G4结合蛋白如何调节G4的稳定性很重要。这项工作提出了一种使用磁镊操纵单个G4分子的方法，该方法能够实时研究G4结合蛋白对单个G4分子的调控。一般来说，该方法适用于蛋白质/配体相互作用以及对各种DNA或RNA二级结构调控研究中的广泛应用。
