Inducing and modulating anisotropic DNA bends by pseudocomplementary peptide nucleic acids.
Inducing and modulating anisotropic DNA bends by pseudocomplementary peptide nucleic acids.
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通过伪互补肽核酸诱导和调节各向异性 DNA 弯曲。
DOI：
10.1073/pnas.0308756101
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发表时间：
2004
期刊：
影响因子：
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通讯作者：
Frank-Kamenetskii,MaximD
中科院分区：
文献类型：
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作者：
Kuhn,Heiko;Cherny,DmitryI;Demidov,VadimV;Frank-Kamenetskii,MaximD
DNA弯曲对细胞内各种DNA功能具有重要意义。在这里，我们证明了假互补肽核酸（pcPNAs）代表了一类通用的，序列特异性dna弯曲剂。凝胶电泳分相分析显示了pcPNAs诱导的各向异性DNA弯曲的发生。DNA弯曲的大小是由圆形排列测定法和电子显微镜确定的，两种方法之间的计算平均值有很好的一致性。一对10-聚合的pcpna与目标DNA序列结合会导致中等程度的DNA弯曲，平均值为40-45°，而一个自聚合的8-聚合PNA与目标DNA结合会产生更大的诱导DNA弯曲平均值。当pcPNA靶位点被常规双链DNA的半整数螺旋转距离分开时，发现这两个弯曲是相的，导致DNA弯曲增强，其平均值在80-90°范围内。DNA双螺旋结构中出现这样一个尖锐的弯曲被证实，并通过两个弯曲DNA片段的二聚化有效地形成170 bp长的DNA小环来利用。与先前设计的非天然DNA弯曲剂相比，pcpna具有两个主要优势：它们在靶向双链DNA时具有非常轻微的序列限制，并且由于它们的结合遵循互补原则，它们可以很容易地设计为选定的目标序列。我们得出的结论是，pcPNAs是在几乎任何选择的位点诱导DNA弯曲的有前途的工具。
DNA bending is significant for various DNA functions in the cell. Here, we demonstrate that pseudocomplementary peptide nucleic acids (pcPNAs) represent a class of versatile, sequence-specific DNA-bending agents. The occurrence of anisotropic DNA bends induced by pcPNAs is shown by gel electrophoretic phasing analysis. The magnitude of DNA bending is determined by circular permutation assay and by electron microscopy, with good agreement of calculated mean values between both methods. Binding of a pair of 10-meric pcPNAs to its target DNA sequence results in moderate DNA bending with a mean value of 40–45°, while binding of one self-pc 8-mer PNA to target DNA yields a somewhat larger average value of the induced DNA bend. Both bends are found to be in phase when the pcPNA target sites are separated by distances of half-integer numbers of helical turns of regular duplex DNA, resulting in an enhanced DNA bend with an average value in the range of 80–90°. The occurrence of such a sharp bend within the DNA double helix is confirmed and exploited through efficient formation of 170-bp-long DNA minicircles by means of dimerization of two bent DNA fragments. The pcPNAs offer two main advantages over previously designed classes of nonnatural DNA-bending agents: they have very mild sequence limitations while targeting duplex DNA and they can easily be designed for a chosen target sequence, because their binding obeys the principle of complementarity. We conclude that pcPNAs are promising tools for inducing bends in DNA at virtually any chosen site.