Apurinic/apyrimidinic (AP) site recognition by the 5′-dRP/AP lyase in poly(ADP-ribose) polymerase-1 (PARP-1)
Apurinic/apyrimidinic (AP) site recognition by the 5′-dRP/AP lyase in poly(ADP-ribose) polymerase-1 (PARP-1)
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DOI：
10.1073/pnas.1009182107
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发表时间：
2010-12-21
影响因子：
11.1
通讯作者：
Lavrik, O. I.
中科院分区：
文献类型：
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作者：
Khodyreva, S. N.;Prasad, R.;Lavrik, O. I.
人聚（ADP-核糖）聚合酶-1（PARP-1）与DNA中完整的脱嘌呤/脱嘧啶（AP）位点相互作用的能力已被证明。在细胞提取物中，PARP-1/AP位点DNA复合物的硼氢化钠还原导致PARP-1与DNA共价交联;通过质谱法确认了交联PARP-1的身份。使用纯化的人PARP-1，在竞争结合实验中证实了PARP-1与含AP位点的DNA结合的特异性。PARP-1在与含AP位点的DNA结合后仅被弱激活以进行聚（ADP-核糖）合成，但在被AP内切核酸酶1（APE 1）切割链后被强激活以进行聚（ADP-核糖）合成。由于其与AP位点的结合，PARP-1可以在碱基切除修复中发挥作用，等待APE 1的DNA链切割或PARP-1中的5 '-dRP/AP裂解酶活性。
The capacity of human poly(ADP-ribose) polymerase-1 (PARP-1) to interact with intact apurinic/apyrimidinic (AP) sites in DNA has been demonstrated. In cell extracts, sodium borohydride reduction of the PARP-1/AP site DNA complex resulted in covalent cross-linking of PARP-1 to DNA; the identity of cross-linked PARP-1 was confirmed by mass spectrometry. Using purified human PARP-1, the specificity of PARP-1 binding to AP site-containing DNA was confirmed in competition binding experiments. PARP-1 was only weakly activated to conduct poly(ADP-ribose) synthesis upon binding to AP site-containing DNA, but was strongly activated for poly (ADP-ribose) synthesis upon strand incision by AP endonuclease 1 (APE1). By virtue of its binding to AP sites, PARP-1 could be poised for its role in base excision repair, pending DNA strand incision by APE1 or the 5'-dRP/AP lyase activity in PARP-1.