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离子对人端粒DNA多态性、有效电荷及稳定性的影响。光子相关光谱和圆二色性研究。

Effect of ions on the polymorphism, effective charge, and stability of human telomeric DNA. Photon correlation spectroscopy and circular dichroism studies.

作者信息

Włodarczyk Agnieszka, Grzybowski Przemysław, Patkowski Adam, Dobek Andrzej

机构信息

Faculty of Physics, Molecular Biopysics Division, A. Mickiewicz University, Umultowska 85, 61-614 Poznań, Poland.

出版信息

J Phys Chem B. 2005 Mar 3;109(8):3594-605. doi: 10.1021/jp045274d.

Abstract

The effect of different ions on the formation and behavior of quadruplex structures of the human telomere sequence d(TTAGGG)(4) has been studied by photon correlation spectroscopy (PCS) and circular dichroism (CD). The saturation and melting curves obtained in the presence of K(+), Na(+), Rb(+), Li(+), Cs(+), and Sr(2+) ions were recorded by CD spectroscopy and indicated the formation of monomeric quadruplexes. Analysis of the saturation curves obtained at 2 degrees C has shown that the presence of a single Sr(2+) ion per oligomer is sufficient for the formation of a monomeric quadruplex of the DNA sequence studied. In the presence of SrCl(2) at a concentration of 50 mM, the formation of tetrameric quadruplexes has been detected. The effect of Sr(2+) ions on the formation of quadruplex structures by the human telomere sequence d(TTAGGG)(4) is stronger and different from that of the other ions tested. The paper also presents results of a study of electrostatic interactions in solution. The translation diffusion coefficients D(T) of the structures present in solution have been determined by photon correlation spectroscopy and the effective charges on the structures have been calculated by combining the experimental data with the results based on the coupled mode theory. Analysis of the melting points monitored by the CD method has permitted a determination of Deltan, the number of ions released in the process of thermal denaturation. All the results are in good agreement with the predictions based on the theory of polyelectrolytes. The effect of ions on the formation and behavior of quadruplex structures of the human telomere sequence d(TTAGGG)(4) has been studied by photon correlation spectroscopy and circular dichroism.

摘要

通过光子相关光谱法(PCS)和圆二色性(CD)研究了不同离子对人类端粒序列d(TTAGGG)(4)四链体结构形成和行为的影响。在存在K(+)、Na(+)、Rb(+)、Li(+)、Cs(+)和Sr(2+)离子的情况下,通过CD光谱记录了饱和曲线和熔解曲线,结果表明形成了单体四链体。对在2℃下获得的饱和曲线的分析表明,每个寡聚物存在单个Sr(2+)离子就足以形成所研究DNA序列的单体四链体。在浓度为50 mM的SrCl(2)存在下,检测到了四聚体四链体的形成。Sr(2+)离子对人类端粒序列d(TTAGGG)(4)四链体结构形成的影响比其他测试离子更强且不同。本文还展示了溶液中静电相互作用的研究结果。通过光子相关光谱法测定了溶液中存在的结构的平移扩散系数D(T),并通过将实验数据与基于耦合模式理论的结果相结合,计算了结构上的有效电荷。通过CD方法监测熔点的分析允许确定Δn,即热变性过程中释放的离子数。所有结果与基于聚电解质理论的预测高度吻合。通过光子相关光谱法和圆二色性研究了离子对人类端粒序列d(TTAGGG)(4)四链体结构形成和行为的影响。

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