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两种不同的构象状态定义了人源 RAD51-ATP 与单链 DNA 的相互作用。

Two distinct conformational states define the interaction of human RAD51-ATP with single-stranded DNA.

机构信息

Department of Physics and Astronomy and LaserLaB, Vrije Universiteit Amsterdam, Amsterdam, The Netherlands.

Department of Biochemistry, University of Cambridge, Cambridge, UK.

出版信息

EMBO J. 2018 Apr 3;37(7). doi: 10.15252/embj.201798162. Epub 2018 Mar 5.

DOI:10.15252/embj.201798162
PMID:29507080
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC5881629/
Abstract

An essential mechanism for repairing DNA double-strand breaks is homologous recombination (HR). One of its core catalysts is human RAD51 (hRAD51), which assembles as a helical nucleoprotein filament on single-stranded DNA, promoting DNA-strand exchange. Here, we study the interaction of hRAD51 with single-stranded DNA using a single-molecule approach. We show that ATP-bound hRAD51 filaments can exist in two different states with different contour lengths and with a free-energy difference of ~4 kT per hRAD51 monomer. Upon ATP hydrolysis, the filaments convert into a disassembly-competent ADP-bound configuration. In agreement with the single-molecule analysis, we demonstrate the presence of two distinct protomer interfaces in the crystal structure of a hRAD51-ATP filament, providing a structural basis for the two conformational states of the filament. Together, our findings provide evidence that hRAD51-ATP filaments can exist in two interconvertible conformational states, which might be functionally relevant for DNA homology recognition and strand exchange.

摘要

修复 DNA 双链断裂的一个基本机制是同源重组 (HR)。其核心催化剂之一是人 RAD51（hRAD51），它在单链 DNA 上组装成螺旋核蛋白丝，促进 DNA 链交换。在这里，我们使用单分子方法研究了 hRAD51 与单链 DNA 的相互作用。我们表明，结合 ATP 的 hRAD51 丝可以存在于两种不同的状态，具有不同的轮廓长度，并具有每个 hRAD51 单体约 4 kT 的自由能差异。在 ATP 水解后，细丝转化为具有解组装能力的 ADP 结合构象。与单分子分析一致，我们在 hRAD51-ATP 细丝的晶体结构中证明了两种不同的单体界面的存在，为细丝的两种构象状态提供了结构基础。总之，我们的发现提供了证据表明 hRAD51-ATP 细丝可以存在于两种可相互转化的构象状态，这对于 DNA 同源性识别和链交换可能具有功能相关性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3607/5881629/69631f46cc66/EMBJ-37-e98162-g012.jpg
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