The excluded DNA strand is SEW important for hexameric helicase unwinding
The excluded DNA strand is SEW important for hexameric helicase unwinding
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DOI：
10.1016/j.ymeth.2016.04.008
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发表时间：
2016-10-01
期刊：
影响因子：
4.8
通讯作者：
Trakselis, Michael A.
中科院分区：
文献类型：
--
作者：
Carney, Sean M.;Trakselis, Michael A.
解旋酶被提出主要通过在一条链上移位以空间上排除和分离两条链来解旋dsDNA。六聚体解旋酶特别地已经显示包围一条链而物理地排除另一条链。在这篇文章中，我们将详细的实验方法，用于验证特定的相互作用与排除链外表面的六聚体解旋酶。定性和定量的方法来确定排除链相互作用，确定外部相互作用的残基，并测量结合的动力学。与nontranslocating链外部相互作用的影响进行了讨论，包括正向解旋稳定，调节解旋速率，和DNA损伤传感。(C)2016 Elsevier Inc. All rights reserved.
Helicases are proposed to unwind dsDNA primarily by translocating on one strand to sterically exclude and separate the two strands. Hexameric helicases in particular have been shown to encircle one strand while physically excluding the other strand. In this article, we will detail experimental methods used to validate specific interactions with the excluded strand on the exterior surface of hexameric helicases. Both qualitative and quantitative methods are described to identify an excluded strand interaction, determine the exterior interacting residues, and measure the dynamics of binding. The implications of exterior interactions with the nontranslocating strand are discussed and include forward unwinding stabilization, regulation of the unwinding rate, and DNA damage sensing. (C) 2016 Elsevier Inc. All rights reserved.