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首页> 外文期刊>Biochemistry >Determining the DNA bending angle induced by non-specific high mobility group-1 (HMG-1) proteins: a novel method.
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Determining the DNA bending angle induced by non-specific high mobility group-1 (HMG-1) proteins: a novel method.

机译：确定非特异性高迁移率族-1（HMG-1）蛋白诱导的DNA弯曲角度：一种新方法。

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摘要

To study the DNA bending induced by non-sequence-specific HMG-1 domain proteins, we have engineered a fusion protein linking the yeast NHP6A with a sequence-specific DNA binding domain, the DNA binding domain of the Hin recombinase, Hin-DBD. A series of biochemical experiments were carried out to characterize the DNA binding property of this fusion protein. Our data showed that the fusion protein not only specifically recognizes a DNA fragment containing the Hin-DBD binding site, but also binds DNA with a higher affinity in comparison with either domain alone. Both domains of the fusion protein are bound to the DNA in juxtaposition. Permutation assays showed that the fusion protein induced a DNA bending at the site of NHP6A binding by an estimated value of 63 degrees. We believe that this experimental design provides an effective vehicle to determine the DNA bending induced by nonspecific HMG-1 proteins.
机译：为了研究由非序列特异性HMG-1域蛋白诱导的DNA弯曲，我们设计了一种融合蛋白，将酵母NHP6A与序列特异性DNA结合域（Hin重组酶Hin-DBD的DNA结合域）连接在一起。进行了一系列的生化实验以表征该融合蛋白的DNA结合特性。我们的数据表明，该融合蛋白不仅能特异性识别含有Hin-DBD结合位点的DNA片段，而且与单独的任一结构域相比都具有更高的亲和力。融合蛋白的两个结构域并列结合到DNA。排列分析表明，融合蛋白在NHP6A结合位点诱导DNA弯曲，估计值为63度。我们认为，该实验设计提供了一种有效的手段来确定由非特异性HMG-1蛋白诱导的DNA弯曲。

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