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首页> 外文期刊>Journal of Molecular Biology >The isomerization of the UvrB-DNA preincision complex couples the UvrB and UvrC activities.
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The isomerization of the UvrB-DNA preincision complex couples the UvrB and UvrC activities.

机译：UvrB-DNA预切割复合体的异构化使UvrB和UvrC活性耦合。

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摘要

In Escherichia coli nucleotide excision repair, the UvrB-DNA preincision complex plays a key role, linking adduct recognition to incision. We previously showed that the efficiency of the incision is inversely related to the stability of the preincision complex. We postulated that an isomerization reaction converts [UvrB-DNA], stable but incompetent for incision, into the [UvrB-DNA]' complex, unstable and competent for incision. Here, we identify two parameters, negative supercoiling and presence of a nick at the fifth phosphodiester bond 3' to the lesion, that accelerate the isomerization leading to an increasing incision efficiency. We also show that the [UvrB-DNA] complex is more resistant to a salt concentration increase than the [UvrB-DNA]' complex. Finally, we report that the [UvrB-DNA]' is recognized by UvrC. These data suggest that the isomerization reaction leads to an exposure of single-stranded DNA around the lesion. This newly exposed single-stranded DNA serves as a binding site and substrate for the UvrC endonuclease. We propose that the isomerization reaction is responsible for coupling UvrB and UvrC activities and that this reaction corresponds to the binding of ATP.
机译：在大肠杆菌核苷酸​​切除修复中，UvrB-DNA切除前复合体发挥关键作用，将加合物识别与切口联系起来。我们先前表明，切口的效率与切口前复合物的稳定性成反比。我们假设异构化反应将稳定但不适合切口的[UvrB-DNA]转换为不稳定且适合切口的[UvrB-DNA]'复合物。在这里，我们确定了两个参数，负超卷和在病变的第五个磷酸二酯键3'处存在切口，这些参数加速了异构化作用，从而提高了切口效率。我们还显示[UvrB-DNA]复合物比[UvrB-DNA]'复合物对盐浓度的增加更具有抵抗力。最后，我们报告[UvrB-DNA]'被UvrC识别。这些数据表明异构化反应导致病变周围单链DNA暴露。这种新暴露的单链DNA充当UvrC核酸内切酶的结合位点和底物。我们建议异构化反应负责耦合UvrB和UvrC活性，并且该反应对应于ATP的结合。

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