Division of Pharmacology and Toxicology, College of Pharmacy, The University of Texas at Austin, Dell Pediatric Research Institute, Austin, TX, USA.
Methods Mol Biol. 2023;2651:227-240. doi: 10.1007/978-1-0716-3084-6_16.
Alternative DNA structures that differ from the canonical B-DNA double helix, including Z-DNA, have received much attention recently due to their impact on DNA metabolic processes, including replication, transcription, and genome maintenance. Non-B-DNA-forming sequences can also stimulate genetic instability associated with disease development and evolution. Z-DNA can stimulate different types of genetic instability events in different species, and several different assays have been established to detect Z-DNA-induced DNA strand breaks and mutagenesis in prokaryotic and eukaryotic systems. In this chapter, we will introduce some of these methods including Z-DNA-induced mutation screening and detection of Z-DNA-induced strand breaks in mammalian cells, yeast, and mammalian cell extracts. Results from these assays should provide better insight into the mechanisms of Z-DNA-related genetic instability in different eukaryotic model systems.
近年来,与经典的 B-DNA 双螺旋结构不同的其他 DNA 结构,包括 Z-DNA,因其对 DNA 代谢过程(包括复制、转录和基因组维持)的影响而受到广泛关注。非 B-DNA 形成序列也会刺激与疾病发生和进化相关的遗传不稳定性。Z-DNA 可以在不同物种中刺激不同类型的遗传不稳定性事件,并且已经建立了几种不同的测定方法来检测原核和真核系统中 Z-DNA 诱导的 DNA 链断裂和突变。在本章中,我们将介绍其中的一些方法,包括 Z-DNA 诱导的突变筛选和检测哺乳动物细胞、酵母和哺乳动物细胞提取物中 Z-DNA 诱导的链断裂。这些测定方法的结果将为不同真核模型系统中与 Z-DNA 相关的遗传不稳定性机制提供更好的见解。