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作者信息

Dubrovin Evgeniy V

机构信息

Lomonosov Moscow State University, Leninskie Gory 1 Bld. 2, 119991 Moscow, Russian Federation.

Moscow Institute of Physics and Technology, Institutskiy Per. 9, Dolgoprudny, 141700 Russian Federation.

出版信息

Biophys Rev. 2023 Aug 26;15(5):1015-1033. doi: 10.1007/s12551-023-01111-3. eCollection 2023 Oct.

Abstract

The interaction of nucleic acids with proteins plays an important role in many fundamental biological processes in living cells, including replication, transcription, and translation. Therefore, understanding nucleic acid-protein interaction is of high relevance in many areas of biology, medicine and technology. During almost four decades of its existence atomic force microscopy (AFM) accumulated a significant experience in investigation of biological molecules at a single-molecule level. AFM has become a powerful tool of molecular biology and biophysics providing unique information about properties, structure, and functioning of biomolecules. Despite a great variety of nucleic acid-protein systems under AFM investigations, there are a number of typical approaches for such studies. This review is devoted to the analysis of the typical AFM-based approaches of investigation of DNA (RNA)-protein complexes with a major focus on transcription studies. The basic strategies of AFM analysis of nucleic acid-protein complexes including investigation of the products of DNA-protein reactions and real-time dynamics of DNA-protein interaction are categorized and described by the example of the most relevant research studies. The described approaches and protocols have many universal features and, therefore, are applicable for future AFM studies of various nucleic acid-protein systems.

摘要

核酸与蛋白质的相互作用在活细胞的许多基本生物学过程中起着重要作用,包括复制、转录和翻译。因此,了解核酸-蛋白质相互作用在生物学、医学和技术的许多领域都具有高度相关性。在其存在的近四十年里,原子力显微镜(AFM)在单分子水平研究生物分子方面积累了丰富的经验。AFM已成为分子生物学和生物物理学的强大工具,可提供有关生物分子特性、结构和功能的独特信息。尽管AFM研究的核酸-蛋白质系统种类繁多,但此类研究仍有一些典型方法。本综述致力于分析基于AFM的研究DNA(RNA)-蛋白质复合物的典型方法,主要侧重于转录研究。通过最相关研究的实例,对核酸-蛋白质复合物AFM分析的基本策略进行了分类和描述,包括对DNA-蛋白质反应产物的研究以及DNA-蛋白质相互作用的实时动力学。所描述的方法和方案具有许多通用特征,因此适用于未来对各种核酸-蛋白质系统的AFM研究。

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