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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >RecA polymerization on double-stranded DNA by using single-molecule manipulation: The role of ATP hydrolysis
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RecA polymerization on double-stranded DNA by using single-molecule manipulation: The role of ATP hydrolysis

机译：通过单分子操作在双链DNA上进行RecA聚合：ATP水解的作用

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摘要

The polymerization of RecA on individual double-strtanded DNA 48.5 Kb), anchored at one end to a cover glass and at the other end to an optically trapped 3-muM diameter polystyrene bead, serves as a template. The elongation caused by RecA assembly is measured in the presence of ATP and ATP[rS]. By using force extension and hydrodynamic recoil, a value of the persistence length of the RecA-DNA complex is obtained. In the presence of ATP, the polymer length is unstable, first growing to saturation and then decreasing. This suggests a transient dynamics of association and dissociation for RecA on a double-stranded DNA, the process being controlled by ATP hydrolysis. Part of this dynamics is suppressed in the presence of ATP[rS], leading to a stabilized RecA-DNA complex. A one-dimensional nucleation and growth model is presented that may account for the protein assembly.
机译：RecA在单个双链DNA（48.5 Kb）上的聚合反应用作模板，该聚合反应的一端固定在防护玻璃上，另一端固定在光学捕获的直径为3μM的聚苯乙烯珠上。在ATP和ATP [rS]的存在下测量由RecA组装引起的伸长。通过使用力的延伸和流体动力学的反冲力，获得了RecA-DNA复合体的持久长度值。在ATP的存在下，聚合物长度不稳定，先增长到饱和，然后下降。这表明RecA在双链DNA上的缔合和解离的瞬时动力学，该过程受ATP水解的控制。在ATP [rS]的存在下，部分动力学被抑制，导致稳定的RecA-DNA复合物。提出了可以解释蛋白质组装的一维成核和生长模型。

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