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首页> 外文期刊>Journal of Molecular Biology >The complex of DNA gyrase and quinolone drugs on DNA forms a barrier to the T7 DNA polymerase replication complex
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The complex of DNA gyrase and quinolone drugs on DNA forms a barrier to the T7 DNA polymerase replication complex

机译：DNA回旋酶和喹诺酮药物在DNA上的复合物对T7 DNA聚合酶复制复合物形成了障碍

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摘要

Quinolone drugs can inhibit bacterial DNA replication, via interaction with the type II topoisomerase DNA gyrase. Using a DNA template containing a preferred site for quinolone-induced gyrase cleavage, we have demonstrated that the passage of the bacteriophage T7 replication complex is blocked in vitro by the formation of a gyrase-drug-DNA complex. The majority of the polymerase is arrested approximately 10 bp upstream of this preferred site, although other minor sites of blocking have been observed. The ability of mutant gyrase proteins to arrest DNA replication in vitro has been investigated. Gyrase containing mutations in the A subunit at either the active-site tyrosine (Tyr122) or Ser83 (a residue known to be involved in quinolone interaction) failed to halt the progress of the polymerase. A low-level, quinolone-resistant mutation in the B subunit of gyrase showed reduced blocking compared to wild-type. We have demonstrated that DNA cleavage and replication blocking occur on similar time-scales and we conclude that formation of the cleavable complex is a prerequisite for polymerase blocking. Addition ally, we have shown that collision of the replication proteins with the gyrase-drug-DNA complex is not sufficient to render this complex irreversible and that further factors must be involved in processing this stalled complex. (C) 2000 Academic Press. [References: 51]
机译：喹诺酮类药物可通过与II型拓扑异构酶DNA旋转酶相互作用来抑制细菌DNA复制。使用含有优选喹诺酮诱导的促旋酶裂解位点的DNA模板，我们已经证明，通过形成促旋酶-药物-DNA复合物在体外阻断了噬菌体T7复制复合物的通过。尽管已观察到其他较小的阻断位点，但大多数聚合酶在该优选位点上游约10 bp处被阻滞。已经研究了突变的促旋酶蛋白在体外阻止DNA复制的能力。活性位点酪氨酸（Tyr122）或Ser83（已知参与喹诺酮相互作用的残基）的A亚基中含有突变的旋涡酶未能阻止聚合酶的进程。与野生型相比，回旋酶B亚基中的一种低水平，喹诺酮抗性突变显示出减少的阻断作用。我们已经证明，DNA裂解和复制阻断发生在相似的时间尺度上，并且我们得出结论，可裂解复合物的形成是聚合酶阻断的先决条件。另外，我们已经表明，复制蛋白与促旋酶-药物-DNA复合物的碰撞不足以使该复合物不可逆，并且在加工该停止的复合物时必须涉及其他因素。 （C）2000学术出版社。 [参考：51]

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