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首页> 外文期刊>Nucleic Acids Research >Inhibition of translesion DNA polymerase by archaeal reverse gyrase.
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Inhibition of translesion DNA polymerase by archaeal reverse gyrase.

机译：古细菌反向回旋酶对跨病变DNA聚合酶的抑制作用。

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摘要

Reverse gyrase is a unique DNA topoisomerase endowed with ATP-dependent positive supercoiling activity. It is typical of microorganisms living at high temperature and might play a role in maintenance of genome stability and repair. We have identified the translesion DNA polymerase SsoPolY/Dpo4 as one partner of reverse gyrase in the hyperthermophilic archaeon Sulfolobus solfataricus. We show here that in cell extracts, PolY and reverse gyrase co-immunoprecipitate with each other and with the single strand binding protein, SSB. The interaction is confirmed in vitro by far-western and Surface Plasmon Resonance. In functional assays, reverse gyrase inhibits PolY, but not the S. solfataricus B-family DNA polymerase PolB1. Mutational analysis shows that inhibition of PolY activity depends on both ATPase and topoisomerase activities of reverse gyrase, suggesting that the intact positive supercoiling activity is required for PolY inhibition. In vivo, reverse gyrase and PolY are degraded after induction of DNA damage. Inhibition by reverse gyrase and degradation might act as a double mechanism to control PolY and prevent its potentially mutagenic activity when undesired. Inhibition of a translesion polymerase by topoisomerase-induced modification of DNA structure may represent a previously unconsidered mechanism of regulation of these two-faced enzymes.
机译：反向旋旋酶是一种独特的DNA拓扑异构酶，具有ATP依赖性正超螺旋活性。它是高温下典型的微生物，可能在维持基因组稳定性和修复中发挥作用。我们已经确定跨病害的DNA聚合酶SsoPolY / Dpo4是在超嗜热古细菌Sulfolobus solfataricus中逆转回旋酶的一种伴侣。我们在这里显示，在细胞提取物中，PolY和反向旋旋酶彼此以及与单链结合蛋白SSB共同免疫沉淀。在远处和表面等离振子共振证实了相互作用。在功能测定中，反向旋回酶抑制PolY，但不能抑制solfataricus B家族DNA聚合酶PolB1。突变分析表明，对PolY活性的抑制取决于反向旋回酶的ATPase和拓扑异构酶活性，这表明PolY抑制需要完整的正超螺旋活性。在体内，DNA损伤诱导后，逆向旋转酶和PolY降解。反向旋转酶的抑制和降解可能是控制PolY并在不希望的时候阻止其潜在诱变活性的双重机制。拓扑异构酶诱导的DNA结构修饰对跨病变聚合酶的抑制作用可能代表了以前尚未考虑的调节这些双面酶的机制。

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