Investigating direct interaction between Escherichia coli topoisomerase I and RecA
Investigating direct interaction between Escherichia coli topoisomerase I and RecA
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DOI：
10.1016/j.gene.2016.03.013
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发表时间：
2016-07-01
期刊：
影响因子：
3.5
通讯作者：
Tse-Dinh, Yuk-Ching
中科院分区：
文献类型：
--
作者：
Banda, Srikanth;Tiwari, Purushottam Babu;Tse-Dinh, Yuk-Ching
蛋白质-蛋白质相互作用在细胞过程中具有特殊的重要性，包括复制、转录、重组和修复。大肠杆菌拓扑异构酶I （EcTOP1）主要参与负DNA超卷曲的松弛。大肠杆菌RecA是同源重组和SOS dna损伤反应的关键蛋白，已被证明可以刺激EcTOP1的松弛活性。它们之间直接的蛋白质-蛋白质相互作用的证据以前还没有得到证实。我们在这里报告了大肠杆菌RecA和拓扑异构酶I之间的直接物理相互作用。我们通过下拉实验和表面等离子体共振测量证明了RecA-拓扑异构酶I之间的相互作用。分子对接支持这样的观察，即相互作用涉及到形成活性位点的拓扑异构酶I n端结构域。我们的下拉实验结果表明，ATP虽然不是必需的，但却增强了RecA-EcTOP1的相互作用。我们提出大肠杆菌RecA与拓扑异构酶I物理相互作用以调节染色体DNA超卷曲。(C) 2016 Elsevier B.V.版权所有
Protein-protein interactions are of special importance in cellular processes, including replication, transcription, recombination, and repair. Escherichia coli topoisomerase I (EcTOP1) is primarily involved in the relaxation of negative DNA supercoiling. E. coli RecA, the key protein for homologous recombination and SOS DNA-damage response, has been shown to stimulate the relaxation activity of EcTOP1. The evidence for their direct protein-protein interaction has not been previously established. We report here the direct physical interaction between E. coli RecA and topoisomerase I. We demonstrated the RecA-topoisomerase I interaction via pull-down assays, and surface plasmon resonance measurements. Molecular docking supports the observation that the interaction involves the topoisomerase I N-terminal domains that form the active site. Our results from pull-down assays showed that ATP, although not required, enhances the RecA-EcTOP1 interaction. We propose that E. coli RecA physically interacts with topoisomerase I to modulate the chromosomal DNA supercoiling. (C) 2016 Elsevier B.V. All rights reserved.