通过单分子荧光共振能量转移显微镜观察G-四链体与蛋白质的结合
G-Quadruplex and Protein Binding by Single-Molecule FRET Microscopy.
作者信息
Lee Chun-Ying, McNerney Christina, Myong Sua
机构信息
Department of Biophysics, Johns Hopkins University, Baltimore, MD, USA.
Department of Biology, Johns Hopkins University, Baltimore, MD, USA.
出版信息
Methods Mol Biol. 2019;2035:309-322. doi: 10.1007/978-1-4939-9666-7_18.
Abstract
G-quadruplex (G4) is a non-canonical nucleic acid structure that arises from the stacking of planar G-tetrads, stabilized by monovalent cations. G4 forming sequences exist throughout the genome and G4 structures are shown to be involved in many processes including DNA replication and gene expression. The single-molecule total internal reflection fluorescence (TIRF) microscopy has been employed to study G4 structure formation and protein binding interactions. Here, we describe methods by which we tested the folding and unfolding of G-quadruplexes structure and studied the dynamics of its interaction with POT1 protein. The methods presented here can be applied to study other putative G4 sequences and potential binding partners.
摘要
G-四链体（G4）是一种非经典核酸结构，由平面G-四联体堆积而成，通过单价阳离子稳定。形成G4的序列存在于整个基因组中，并且G4结构被证明参与包括DNA复制和基因表达在内的许多过程。单分子全内反射荧光（TIRF）显微镜已被用于研究G4结构的形成和蛋白质结合相互作用。在这里，我们描述了测试G-四链体结构折叠和展开并研究其与POT1蛋白相互作用动力学的方法。这里介绍的方法可应用于研究其他假定的G4序列和潜在的结合伴侣。
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