Single molecule force spectroscopy on G-quadruplex DNA.
Single molecule force spectroscopy on G-quadruplex DNA.
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DOI：
10.1002/chem.200901390
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发表时间：
2009-08-17
影响因子：
4.3
通讯作者：
Sinniah, Kumar
中科院分区：
文献类型：
--
作者：
Lynch, Susanna;Baker, Heather;Byker, Sarah G.;Zhou, Dejian;Sinniah, Kumar
在过去的十年里，人们对鸟嘌呤(G)四链体的兴趣与日俱增，因为它们存在于人类端粒、控制基因稳定性的染色体末端以及基因组的其他部分，尤其是启动子中。[3]它们已被证明抑制端粒酶的活性，端粒酶是一种维持端粒DNA适当长度的酶，在许多类型的癌症中过度表达。[4]因此，G-四链体DNA一直是癌症治疗的一个有吸引力的靶点。[5]尽管人们对G-四链体的拓扑结构很感兴趣，它们的物理性质还没有被很好地理解。[6]在溶液中使用荧光和表面等离子体共振(SPR)方法获得的动力学数据之间有很大的差异，可能是由于荧光团标记的不稳定[7a-c]和基于间接杂交的SPR方法中的表面阻碍。此外，由一条DNA链形成的G-四链体可能具有不同的构象[6.7]，其中DNA序列的微小变化可能导致它们的热力学性质的显著差异。[6]通过直接对未标记的单个分子进行测量，一次一个，避免系综平均，基于原子力显微镜(AFM)的单分子(SM)力光谱(FS)已被证明是了解广泛的生物学问题的有力工具：蛋白质折叠，[8]蛋白质-配体相互作用，[9]和DNA碱基配对。[10]考虑到大量可能的G-四链拓扑，[1.2]SM-FS可能为它们的结构、功能和稳定性提供独特的见解。此外，最近对SM方法的理论分析的发展使得对动力学和热力学参数的可靠估计成为可能。[11]据我们所知，AFM SMFS还没有被用于研究G-四链。[12]在这里，我们提出了第一个基于AFM的双分子G-四链系统的SMFS研究。
Interest in guanine (G) quadruplexes [1] has intensified over the past decade because they are found in human telomeres,[2] the chromosome ends that govern gene stability, and other parts of the genome, especially in promoters.[3] They have been shown to inhibit the activity of telomerase, an enzyme which maintains the proper length of telomere DNAs and is overexpressed in many types of cancer.[4] As such, the G-quadruplex DNA has been an attractive target for cancer therapy.[5] Despite the wide interest in the topology of G-quadruplexes, their physical properties are not well-understood.[6] There are large differences between the kinetic data obtained in solution using fluorescence and on surfaces using surface plasmon resonance (SPR) approaches,[7] possibly due to destabilization by fluorophore labelling [7a-c] and surface hindrance in the indirect hybridization based SPR approach.[7d] The kinetics and thermodynamics are expected to be different for G-quadruplexes formed from uni-, bi-, and tetra-molecular DNA strands. Furthermore, G-quadruplexes formed from a single DNA strand may have different conformations,[6.7] where minor changes in the DNA sequence may result in significant differences in their thermodynamic properties.[6]By taking measurements directly on unlabelled individual single molecules, one at a time, avoiding ensemble averaging, single-molecule (SM) force spectroscopy (FS) based on atomic force microscopy (AFM) has been demonstrated as a powerful tool to gain insights to a wide range of biological problems: protein folding,[8] protein-ligand interactions,[9] and DNA base pairing.[10] Given the large number of G-quadruplex topologies possible,[1.2] SM-FS can potentially provide unique insights into their structure, function, and stability. Further, the recent development of theoretical analysis for SM approaches has enabled reliable estimates of the kinetic and thermodynamic parameters.[11] To the best of our knowledge, the AFM SMFS has not been used to study G-quadruplex.[12] Herein, we present the first AFM based SMFS study on a bi-molecular G-quadruplex system.
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DOI：
10.1073/pnas.0806085105
发表时间：
2008-10-14
影响因子：
11.1
作者：
Dudko, Olga K.;Hummer, Gerhard;Szabo, Attila
通讯作者：
Szabo, Attila
影响因子：
3.9
作者：
Wal, MV;Kamper, S;Sinniah, K
通讯作者：
Sinniah, K
影响因子：
14.9
作者：
Lane AN;Chaires JB;Gray RD;Trent JO
通讯作者：
Trent JO
影响因子：
2
作者：
Pope, LH;Davies, MC;Williams, PM
通讯作者：
Williams, PM
影响因子：
3.9
作者：
Neidle, Stephen;Parkinson, Gary N.
通讯作者：
Parkinson, Gary N.