Structural diversity of target-specific homopyrimidine peptide nucleic acid-dsDNA complexes.

Structural diversity of target-specific homopyrimidine peptide nucleic acid-dsDNA complexes.
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DOI:
10.1093/nar/gkl736
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发表时间:
2006
影响因子:
14.9
通讯作者:
Nielsen PE
Nielsen PE
中科院分区:
生物学2区
文献类型:
--
作者:
Bentin T;Hansen GI;Nielsen PE

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同嘧啶肽核酸(PNA)寡聚物对双链(ds)DNA的序列选择性识别可通过大沟三链体结合或通过三链体P -环形成的螺旋侵入发生。我们使用凝胶迁移和化学探测分析比较了一个十聚体、一个十二聚体和一个十五聚体的胸腺嘧啶 - 胞嘧啶同嘧啶PNA寡聚物与双链DNA中一个序列互补的同嘌呤靶标的结合。我们发现所有这三种PNA都能形成稳定的三链体侵入复合物,以及与dsDNA靶标形成常规的三链体。三链体形成的动力学比侵入复合物快得多,并且在较低的PNA浓度和较短的孵育时间下占优势。此外,增加离子强度强烈有利于三链体形成而非侵入,因为后者受到阳离子的严重抑制。十聚体PNA形成单一的三链体侵入复合物,而十二聚体PNA有两种结构不同的靶标特异性侵入复合物被鉴定出来,十五聚体PNA则有五种以上。最后,研究表明,孤立的三链体复合物可以在不使Hoogsteen碱基配对的三链体PNA解离的情况下转化为特定的侵入复合物。这些结果清楚地展示了PNA螺旋侵入的“三链体优先”机制的一个实例。
Sequence-selective recognition of double-stranded (ds) DNA by homopyrimidine peptide nucleic acid (PNA) oligomers can occur by major groove triplex binding or by helix invasion via triplex P-loop formation. We have compared the binding of a decamer, a dodecamer and a pentadecamer thymine–cytosine homopyrimidine PNA oligomer to a sequence complementary homopurine target in duplex DNA using gel-shift and chemical probing analyses. We find that all three PNAs form stable triplex invasion complexes, and also conventional triplexes with the dsDNA target. Triplexes form with much faster kinetics than invasion complexes and prevail at lower PNA concentrations and at shorter incubation times. Furthermore, increasing the ionic strength strongly favour triplex formation over invasion as the latter is severely inhibited by cations. Whereas a single triplex invasion complex is formed with the decameric PNA, two structurally different target-specific invasion complexes were characterized for the dodecameric PNA and more than five for the pentadecameric PNA. Finally, it is shown that isolated triplex complexes can be converted to specific invasion complexes without dissociation of the Hoogsteen base-paired triplex PNA. These result demonstrate a clear example of a ‘triplex first’ mechanism for PNA helix invasion.
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发表时间: 1992-11-27
期刊: SCIENCE
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