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首页> 外文期刊>Nucleic Acids Research >DNA linking number change induced by sequence-specific DNA-binding proteins
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DNA linking number change induced by sequence-specific DNA-binding proteins

机译：序列特异性DNA结合蛋白诱导的DNA连接数变化

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摘要

Sequence-specific DNA-binding proteins play a key role in many fundamental biological processes, such as transcription, DNA replication and recombination. Very often, these DNA-binding proteins introduce structural changes to the target DNA-binding sites including DNA bending, twisting or untwisting and wrapping, which in many cases induce a linking number change (Delta Lk) to the DNA-binding site. Due to the lack of a feasible approach, Delta Lk induced by sequence-specific DNA-binding proteins has not been fully explored. In this paper we successfully constructed a series of DNA plasmids that carry many tandem copies of a DNA-binding site for one sequence-specific DNA-binding protein, such as lambda O, LacI, GalR, CRP and AraC. In this case, the protein-induced Delta Lk was greatly amplified and can be measured experimentally. Indeed, not only were we able to simultaneously determine the protein-induced Delta Lk and the DNA-binding constant for lambda O and GalR, but also we demonstrated that the protein-induced Delta Lk is an intrinsic property for these sequence-specific DNA-binding proteins. Our results also showed that protein-mediated DNA looping by AraC and LacI can induce a Delta Lk to the plasmid DNA templates. Furthermore, we demonstrated that the protein-induced Delta Lk does not correlate with the protein-induced DNA bending by the DNA-binding proteins.
机译：序列特异性的DNA结合蛋白在许多基本生物学过程中起着关键作用，例如转录，DNA复制和重组。这些DNA结合蛋白通常会向目标DNA结合位点引入结构变化，包括DNA弯曲，扭曲或解旋和包裹，这在许多情况下会导致DNA结合位点的连接数变化（Delta Lk）。由于缺乏可行的方法，尚未完全探索由序列特异性DNA结合蛋白诱导的Delta Lk。在本文中，我们成功地构建了一系列DNA质粒，这些质粒携带了一个序列特异性DNA结合蛋白（例如lambda O，LacI，GalR，CRP和AraC）的DNA结合位点的许多串联拷贝。在这种情况下，蛋白质诱导的Delta Lk被大大扩增，可以通过实验进行测量。实际上，我们不仅能够同时确定蛋白质诱导的Delta Lk以及lambda O和GalR的DNA结合常数，而且我们还证明了蛋白质诱导的Delta Lk是这些序列特异性DNA-结合蛋白。我们的结果还表明，由AraC和LacI介导的蛋白质介导的DNA环化可以诱导质粒DNA模板产生Delta Lk。此外，我们证明了蛋白质诱导的Delta Lk与DNA结合蛋白引起的蛋白质诱导的DNA弯曲不相关。

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