RNA 中 A-Z 连接点被 Zα 结构域结合的方式。
Adoption of A-Z Junctions in RNAs by Binding of Zα Domains.
机构信息
Department of Biochemistry and Molecular Genetics, University of Colorado Denver School of Medicine, Aurora, CO, USA.
Colorado School of Mines, Golden, CO, USA.
出版信息
Methods Mol Biol. 2023;2651:251-275. doi: 10.1007/978-1-0716-3084-6_18.
While DNA and RNA helices often adopt the canonical B- or A-conformation, the fluid conformational landscape of nucleic acids allows for many higher energy states to be sampled. One such state is the Z-conformation of nucleic acids, which is unique in that it is left-handed and has a "zigzag" backbone. The Z-conformation is recognized and stabilized by Z-DNA/RNA binding domains called Zα domains. We recently demonstrated that a wide range of RNAs can adopt partial Z-conformations termed "A-Z junctions" upon binding to Zα and that the formation of such conformations may be dependent upon both sequence and context. In this chapter, we present general protocols for characterizing the binding of Zα domains to A-Z junction-forming RNAs for the purpose of determining the affinity and stoichiometry of interactions as well as the extent and location of Z-RNA formation.
虽然 DNA 和 RNA 螺旋通常采用典型的 B-或 A-构象，但核酸的流体构象景观允许许多更高能量的状态被采样。其中一种状态是核酸的 Z-构象，它是独特的，因为它是左手的，并且具有“锯齿状”的骨架。Z-构象被称为 Zα 结构域的 Z-DNA/RNA 结合结构域识别和稳定。我们最近证明，在与 Zα 结合时，广泛的 RNA 可以采用部分 Z-构象，称为“A-Z 连接”，并且这种构象的形成可能取决于序列和上下文。在本章中，我们提供了表征 Zα 结构域与形成 A-Z 连接的 RNA 结合的一般方案，目的是确定相互作用的亲和力和化学计量、Z-RNA 形成的程度和位置。
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