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首页> 外文期刊>Journal of Biomolecular Structure and Dynamics >Formation and Thermodynamic Stability of Intermolecular (R*R*Y) DNA Triplex In GAA/TTC Repeats Associated with Freidreich s Ataxia.
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Formation and Thermodynamic Stability of Intermolecular (R*R*Y) DNA Triplex In GAA/TTC Repeats Associated with Freidreich s Ataxia.

机译：GAA / TTC中与Freidreich s共济失调相关的分子间（R * R * Y）DNA三链体的形成和热力学稳定性。

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摘要

It is well established that GAA/TTC base triplet expansion is the cause of degenerative disorder in Freidreich s Ataxia. It is also known that these repeat sequences fold back to form the unusual intramolecular triple helix structures in DNA of the type Pyrimidine *Purine *Pyrimidine or Purine *Purine*Pyrimidine. In this paper we report on the stability of Purine *Purine*Pyrimidine intermolecular triple helix DNA containing GAA/TTC repeats under physiological conditions. Using the oligonucleotides 5 TCGC GAA GAA GAA GAA GAA CGCT 3 , 5 -AGCG TTC TTC TTC TTC TTC GCGA-3 for duplex and 5 - AAG AAG AAG AAG AAG -3 as triplex forming strand (TFO), we have established the formation of triplex by UV melting temperature (Tm), stoichiometry of mixing and circular dichroic spectra. This was further confirmed by gel-retardation assay. The thermodynamic parameters DeltaG, DeltaH and DeltaS for both duplex and triplex formation were determined at different salt concentrations. The results suggest the formation of a stable intermolecular, anti - parallel triplex in GAA / TTC repeat sequences where each repeat unit contains two A*A*T and one G*G*C triplet. The therapeutic agents and TFOs, which competitively inhibit the in-vivo intra-molecular triplex by formation of a more stable inter- molecular triplex, could be used to reverse the transcription deficit in GAA/TTC expansions in Frataxin gene.
机译：公认的是，GAA / TTC碱基三联体扩增是弗莱德里希共济失调退化性疾病的原因。还已知这些重复序列折回以在嘧啶*嘌呤*嘧啶或嘌呤*嘌呤*嘧啶类型的DNA中形成不寻常的分子内三螺旋结构。在本文中，我们报告了在生理条件下含有GAA / TTC重复序列的嘌呤*嘌呤*嘧啶分子间三螺旋DNA的稳定性。使用寡核苷酸5 TCGC GAA GAA GAA GAA GAA CGCT 3，用于双链体的5 -AGCG TTC TTC TTC TTC TTC GCGA-3和作为三链体形成链（TFO）的5-AAG AAG AAG AAG AAG -3，我们确定了紫外熔融温度（Tm），混合化学计量和圆二向色光谱三重。凝胶延迟测定进一步证实了这一点。在不同的盐浓度下确定双链和三链形成的热力学参数DeltaG，DeltaH和DeltaS。结果表明在GAA / TTC重复序列中形成了稳定的分子间反平行三链体，其中每个重复单元包含两个A * A * T和一个G * G * C三联体。通过形成更稳定的分子间三联体竞争性抑制体内分子内三联体的治疗剂和TFO可用于逆转Frataxin基因中GAA / TTC扩增中的转录缺陷。

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