Winardhi Ricksen S, Yan Jie
Department of Physics, National University of Singapore, Singapore, 117542, Singapore.
Mechanobiology Institute, National University of Singapore, Singapore, 117411, Singapore.
Methods Mol Biol. 2017;1624:173-191. doi: 10.1007/978-1-4939-7098-8_14.
Nucleoid-associated proteins (NAPs) are important factors in shaping bacterial nucleoid and regulating global gene expression. A great deal of insights into NAPs can be obtained through studies using single DNA molecule, which has been made possible owing to recent rapid development of single-DNA manipulation techniques. These studies provide information on modes of binding to DNA, which shed light on the mechanism underlying the regulatory function of NAPs. In addition, how NAPs organize DNA and thus their contribution to chromosomal DNA packaging can be determined. In this chapter, we introduce transverse magnetic tweezers that allows for convenient manipulation of long DNA molecules, and its applications in studies of NAPs as exemplified by the E. coli H-NS protein. We describe how transverse magnetic tweezers is a powerful tool that can be used to characterize the DNA binding and organization modes of NAPs and how such information leads to better understanding of its roles in DNA packaging of bacterial nucleoid and transcription regulation.
类核相关蛋白(NAPs)是塑造细菌类核及调控全局基因表达的重要因素。通过使用单分子DNA的研究能够获得许多关于NAPs的深刻见解,这得益于近年来单DNA操作技术的迅速发展才得以实现。这些研究提供了与DNA结合模式相关的信息,从而揭示了NAPs调控功能背后的机制。此外,还能够确定NAPs如何组织DNA以及它们对染色体DNA包装的贡献。在本章中,我们介绍了能够方便地操纵长DNA分子的横向磁镊,以及它在以大肠杆菌H-NS蛋白为例的NAPs研究中的应用。我们描述了横向磁镊是如何成为一种强大的工具,可用于表征NAPs的DNA结合和组织模式,以及这些信息如何有助于更好地理解其在细菌类核DNA包装和转录调控中的作用。