Tracking topoisomerase activity at the single-molecule level.

G Charvin, T R Strick, D Bensimon, V Croquette
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引用次数: 92

Abstract

The recent development of new techniques to manipulate single DNA molecules has opened new opportunities for the study of the enzymes that control DNA topology: the type I and II topoisomerases. These single-molecule assays provide a unique way to study the uncoiling of single supercoiled DNA molecules and the unlinking of two intertwined DNAs. They allow for a detailed characterization of the activity of topoisomerases, including the processivity, the chiral discrimination, and the dependence of their enzymatic rate on ATP concentration, degree of supercoiling, and the tension in the molecule. These results shed new light on the mechanism of these enzymes and their function in vivo.

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在单分子水平上跟踪拓扑异构酶活性。
最近,操纵单个DNA分子的新技术的发展为研究控制DNA拓扑结构的酶开辟了新的机会:I型和II型拓扑异构酶。这些单分子分析提供了一种独特的方法来研究单个超卷曲DNA分子的解开和两个缠绕在一起的DNA的解开。它们允许对拓扑异构酶的活性进行详细的表征,包括加工性、手性辨别,以及它们的酶促速率对ATP浓度、超卷曲程度和分子张力的依赖。这些结果揭示了这些酶的作用机制及其在体内的功能。
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