一种用于检测G-四链体相互作用化合物的DNA聚合酶终止试验。
A DNA polymerase stop assay for G-quadruplex-interactive compounds.
作者信息
Han H, Hurley L H, Salazar M
机构信息
Program in Molecular Biology, Division of Medicinal Chemistry and Drug Dynamics Institute, College of Pharmacy, The University of Texas at Austin, Austin, TX 78712, USA.
出版信息
Nucleic Acids Res. 1999 Jan 15;27(2):537-42. doi: 10.1093/nar/27.2.537.
We have developed and characterized an assay for G-quadruplex-interactive compounds that makes use of the fact that G-rich DNA templates present obstacles to DNA synthesis by DNA polymerases. Using Taq DNA polymerase and the G-quadruplex binding 2, 6-diamidoanthraquinone BSU-1051, we find that BSU-1051 leads to enhanced arrest of DNA synthesis in the presence of K+by stabilizing an intramolecular G-quadruplex structure formed by four repeats of either TTGGGG or TTAGGG in the template strand. The data provide additional evidence that BSU-1051 modulates telomerase activity by stabilization of telomeric G-quadruplex DNA and point to a polymerase arrest assay as a sensitive method for screening for G-quadruplex-interactive agents with potential clinical utility.
我们开发并表征了一种用于检测与G-四链体相互作用化合物的分析方法，该方法利用富含鸟嘌呤的DNA模板会阻碍DNA聚合酶进行DNA合成这一事实。使用Taq DNA聚合酶和与G-四链体结合的2,6-二氨基蒽醌BSU-1051，我们发现，在存在K+的情况下，BSU-1051通过稳定模板链中由TTGGGG或TTAGGG的四个重复序列形成的分子内G-四链体结构，导致DNA合成的增强阻滞。这些数据提供了额外的证据，表明BSU-1051通过稳定端粒G-四链体DNA来调节端粒酶活性，并指出聚合酶阻滞分析是一种筛选具有潜在临床应用价值的G-四链体相互作用剂的灵敏方法。
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