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Base-stacking and base-pairing contributions into thermal stability of the DNA double helix

机译:碱基堆积和碱基配对对DNA双螺旋热稳定性的贡献

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Two factors are mainly responsible for the stability of the DNA double helix: base pairing between complementary strands and stacking between adjacent bases. By studying DNA molecules with solitary nicks and gaps we measure temperature and salt dependence of the stacking free energy of the DNA double helix. For the first time, DNA stacking parameters are obtained directly (without extrapolation) for temperatures from below room temperature to close to melting temperature. We also obtain DNA stacking parameters for different salt concentrations ranging from 15 to 100 mM Na+. From stacking parameters of individual contacts, we calculate base-stacking contribution to the stability of A center dot T- and G center dot C-containing DNA polymers. We find that temperature and salt dependences of the stacking term fully determine the temperature and the salt dependence of DNA stability parameters. For all temperatures and salt concentrations employed in present study, base-stacking is the main stabilizing factor in the DNA double helix. A center dot T pairing is always destabilizing and G center dot C pairing contributes almost no stabilization. Base-stacking interaction dominates not only in the duplex overall stability but also significantly contributes into the dependence of the duplex stability on its sequence.
机译:两个因素主要负责DNA双螺旋的稳定性:互补链之间的碱基配对和相邻碱基之间的堆积。通过研究具有孤立缺口和缺口的DNA分子,我们测量了DNA双螺旋堆积自由能的温度和盐依赖性。第一次,直接获得了DNA堆积参数(无需外推),其温度范围从低于室温到接近融化温度。我们还获得了15至100 mM Na +范围内不同盐浓度的DNA堆积参数。从各个接触点的堆叠参数,我们计算出碱基堆叠对包含A中心点T和G中心点C的DNA聚合物稳定性的贡献。我们发现堆叠项的温度和盐依赖性完全决定了DNA稳定性参数的温度和盐依赖性。对于本研究中使用的所有温度和盐浓度,碱基堆积是DNA双螺旋结构中的主要稳定因素。中心点T配对始终不稳定,而G中心点C配对几乎没有稳定作用。碱基堆积相互作用不仅在双链体的总体稳定性中占主导地位,而且在双链体稳定性对其序列的依赖性方面也起着重要作用。

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