英语翻译Transcription-coupled DNA repair uses components of the
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英语翻译
Transcription-coupled DNA repair uses components of the transcription machinery to identify DNA lesions and initiate their repair.These repair pathways are complex,so their mechanistic features remain poorly understood.Bacterial transcription-coupled repair is initiated when RNA polymerase stalled at a DNA lesion is removed by Mfd,an ATP-dependent DNA translocase1,2,3.Here we use single-molecule DNA nanomanipulation to observe the dynamic interactions of Escherichia coli Mfd with RNA polymerase elongation complexes stalled by a cyclopyrimidine dimer or by nucleotide starvation.We show that Mfd acts by catalysing two irreversible,ATP-dependent transitions with different structural,kinetic and mechanistic features.Mfd remains bound to the DNA in a long-lived complex that could act as a marker for sites of DNA damage,directing assembly of subsequent DNA repair factors.These results provide a framework for considering the kinetics of transcription-coupled repair in vivo,and open the way to reconstruction of complete DNA repair pathways at single-molecule resolution.
Transcription-coupled DNA repair uses components of the transcription machinery to identify DNA lesions and initiate their repair.These repair pathways are complex,so their mechanistic features remain poorly understood.Bacterial transcription-coupled repair is initiated when RNA polymerase stalled at a DNA lesion is removed by Mfd,an ATP-dependent DNA translocase1,2,3.Here we use single-molecule DNA nanomanipulation to observe the dynamic interactions of Escherichia coli Mfd with RNA polymerase elongation complexes stalled by a cyclopyrimidine dimer or by nucleotide starvation.We show that Mfd acts by catalysing two irreversible,ATP-dependent transitions with different structural,kinetic and mechanistic features.Mfd remains bound to the DNA in a long-lived complex that could act as a marker for sites of DNA damage,directing assembly of subsequent DNA repair factors.These results provide a framework for considering the kinetics of transcription-coupled repair in vivo,and open the way to reconstruction of complete DNA repair pathways at single-molecule resolution.
DNA转录偶联修复使用的转录机器识别DNA损伤,并启动其维修的组件.这些修复途径很复杂,所以机械功能仍知之甚少.RNA聚合酶的DNA病变停滞在启动时的ATP-依赖的DNA反禄ASE1,2,3的MFD,除去细菌转录偶联修复.在这里,我们使用单分子DNA纳米操纵,动态观察MFD停滞的cyclopyrimi用餐二聚体或核苷酸饥饿与RNA聚合酶伸长复合物相互作用的大肠杆菌.我们发现,通过促进两个不可逆的,依赖于ATP的转换具有不同的结构,动力学和机械功能的MFD行为. K.K.制的保持在一个长寿命的复杂程度,作为DNA损伤位点的标记物,指示组件的随后的DNA修复因子的DNA结合.这些结果提供了一个框架,考虑在体内的转录偶联修复的动力学,单分子分辨率重建完整的DNA修复途径,在打开的方式
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