Collins Bridget E, Ye Ling F, Duzdevich Daniel, Greene Eric C
Department of Biological Sciences, Columbia University, New York, USA.
Department of Biochemistry and Molecular Biophysics, Columbia University, New York, USA; Howard Hughes Medical Institute, Columbia University, New York, USA.
Methods Cell Biol. 2014;123:217-34. doi: 10.1016/B978-0-12-420138-5.00012-4.
Interactions between proteins and nucleic acids are at the molecular foundations of most key biological processes, including DNA replication, genome maintenance, the regulation of gene expression, and chromosome segregation. A complete understanding of these types of biological processes requires tackling questions with a range of different techniques, such as genetics, cell biology, molecular biology, biochemistry, and structural biology. Here, we describe a novel experimental approach called "DNA curtains" that can be used to complement and extend these more traditional techniques by providing real-time information about protein-nucleic acid interactions at the level of single molecules. We describe general features of the DNA curtain technology and its application to the study of protein-nucleic acid interactions in vitro. We also discuss some future developments that will help address crucial challenges to the field of single-molecule biology.
蛋白质与核酸之间的相互作用是大多数关键生物过程的分子基础,包括DNA复制、基因组维持、基因表达调控和染色体分离。要全面理解这些类型的生物过程,需要运用一系列不同的技术来解决问题,如遗传学、细胞生物学、分子生物学、生物化学和结构生物学。在这里,我们描述了一种名为“DNA幕帘”的新型实验方法,它可以通过在单分子水平上提供有关蛋白质 - 核酸相互作用的实时信息,来补充和扩展这些更为传统的技术。我们描述了DNA幕帘技术的一般特征及其在体外蛋白质 - 核酸相互作用研究中的应用。我们还讨论了一些未来的发展方向,这些发展将有助于应对单分子生物学领域的关键挑战。