Poly(ADP-ribosyl)ation as a DNA damage-induced post-translational modification regulating poly(ADP-ribose) polymerase-1-topoisomerase I interaction
Poly(ADP-ribosyl)ation as a DNA damage-induced post-translational modification regulating poly(ADP-ribose) polymerase-1-topoisomerase I interaction
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DOI：
10.1074/jbc.m402729200
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发表时间：
2004-09-17
影响因子：
4.8
通讯作者：
Satoh, MS
中科院分区：
文献类型：
--
作者：
Yung, TMC;Sato, S;Satoh, MS
聚（ADP-核糖基）化是在细胞暴露于DNA损伤剂后立即发生的翻译后修饰。在体内，90%的ADP-核糖聚合物连接到聚（ADP-核糖）聚合酶-1（PARP-1）的自动修饰结构域，所述聚（ADP-核糖）聚合酶-1（PARP-1）是催化该修饰反应的主要酶。这种酶与转录起始、DNA复制和DNA修复因子形成复合物。在大多数已知的情况下，相互作用通过自动修饰域发生。然而，这些相互作用的自动修饰反应的功能影响尚未得到阐明。在本研究中，我们创建了荧光蛋白标记的PARP-1来研究活细胞中的这种酶，并专注于PARP-1和拓扑异构酶I（Topo I）之间的相互作用，拓扑异构酶I是体外与PARP-1相互作用的酶之一。在这里，我们证明了PARP-1与拓扑异构酶I在整个细胞周期中共定位。从生物发光共振能量转移分析的结果表明，共定位是因为一个直接的蛋白质-蛋白质相互作用。响应于DNA损伤，观察到PARP-1去定位和生物发光共振能量转移信号的减少，因为自修饰反应，表明自修饰反应导致PARP-1和Topo I之间的相互作用的破坏。因为已经报道PARP-1促进了Topo I活性，所以我们随后研究了这种相互作用的破坏对Topo I活性的影响，并且我们发现这种破坏导致Topo I活性的降低。这些结果表明，自修饰反应的功能是调节PARP-1和Topo I之间的相互作用，从而调节Topo I活性，以响应DNA损伤。
Poly(ADP-ribosyl) ation is a post-translational modification that occurs immediately after exposure of cells to DNA damaging agents. In vivo, 90% of ADP-ribose polymers are attached to the automodification domain of poly( ADP-ribose) polymerase-1 (PARP-1), the main enzyme catalyzing this modification reaction. This enzyme forms complexes with transcription initiation, DNA replication, and DNA repair factors. In most known cases, the interactions occur through the automodification domain. However, functional implications of the automodification reaction on these interactions have not yet been elucidated. In the present study, we created fluorescent protein-tagged PARP-1 to study this enzyme in live cells and focused on the interaction between PARP-1 and topoisomerase I ( Topo I), one of the enzymes that interacts with PARP-1 in vitro. Here, we demonstrate that PARP-1 co-localizes with Topo I throughout the cell cycle. Results from bioluminescence resonance energy transfer assays suggest that the co-localization is because of a direct protein-protein interaction. In response to DNA damage, PARP-1 de-localization and a reduction in bioluminescence resonance energy transfer signal because of the automodification reaction are observed, suggesting that the automodification reaction results in the disruption of the interaction between PARP-1 and Topo I. Because Topo I activity has been reported to be promoted by PARP-1, we then investigated the effect of the disruption of this interaction on Topo I activity, and we found that this disruption results in the reduction of Topo I activity. These results suggest that a function for the automodification reaction is to regulate the interaction between PARP-1 and Topo I, and consequently, the Topo I activity, in response to DNA damage.