首页> 美国卫生研究院文献>Nucleic Acids Research >The checkpoint clamp Rad9-Rad1-Hus1 complex preferentially stimulates the activity of apurinic/apyrimidinic endonuclease 1 and DNA polymerase β in long patch base excision repair
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The checkpoint clamp Rad9-Rad1-Hus1 complex preferentially stimulates the activity of apurinic/apyrimidinic endonuclease 1 and DNA polymerase β in long patch base excision repair

机译:检查点钳位Rad9-Rad1-Hus1复合体在长斑片碱基切除修复中优先刺激嘌呤/嘧啶内切核酸酶1和DNA聚合酶β的活性

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摘要

Growing evidence suggests that the Rad9-Rad1-Hus1 complex (the 9-1-1 complex), besides its functions in DNA damage sensing and signaling pathways, plays also a direct role in various DNA repair processes. Recent studies have demonstrated that the 9-1-1 complex physically and functionally interacts with several components of the base excision repair (BER) machinery namely DNA polymerase >β (Pol >β), flap endonuclease 1 (Fen 1), DNA ligase I (Lig I) and the MutY homologue of Schizosaccharomyces pombe. In this work, we found for the first time that the 9-1-1 complex interacts in vitro and in vivo with the apurinic/apyrimidinic endonuclease 1 (APE 1), an early component of BER, and can stimulate its AP-endonuclease activity. Moreover, we show that the 9-1-1 complex possesses a stimulatory effect on long patch base excision repair (LP-BER) reconstituted in vitro. The enhancement of LP-BER activity is due to the specific stimulation of the two early components of the repair machinery, namely APE 1 and Pol >β, suggesting a hierarchy of interactions between the 9-1-1 complex and the BER proteins acting in the repairosome. Overall, our results indicate that the 9-1-1 complex is directly involved in LP-BER, thus providing a possible link between DNA damage checkpoints and BER.
机译:越来越多的证据表明,Rad9-Rad1-Hus1复合物(9-1-1复合物)除了在DNA损伤传感和信号传导途径中发挥功能外,还在各种DNA修复过程中起着直接作用。最近的研究表明9-1-1复合物在物理和功能上与碱基切除修复(BER)机器的几个组成部分相互作用,即DNA聚合酶>β(Pol >β) ,皮瓣内切核酸酶1(Fen 1),DNA连接酶I(Lig I)和粟酒裂殖酵母的MutY同源物。在这项工作中,我们首次发现9-1-1复合物在体外和体内与BER的早期成分嘌呤/嘧啶内切核酸酶1(APE 1)相互作用,并且可以刺激其AP内切核酸酶活性。 。此外,我们显示9-1-1复合物对体外重建的长斑片基切除修复(LP-BER)具有刺激作用。 LP-BER活性的增强归因于修复机制的两个早期组成部分的特异性刺激,即APE 1和Pol >β,表明9-1-1复合体之间相互作用的层次BER蛋白在修复小体中起作用。总的来说,我们的结果表明9-1-1复合物直接参与LP-BER,因此提供了DNA损伤检查点和BER之间的可能联系。

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