Dynamics of endogenous PARP1 and PARP2 during DNA damage revealed by live-cell single-molecule imaging.
Dynamics of endogenous PARP1 and PARP2 during DNA damage revealed by live-cell single-molecule imaging.
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DOI：
10.1016/j.isci.2022.105779
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发表时间：
2023-01-20
期刊：
影响因子：
5.8
通讯作者：
Luger, Karolin
中科院分区：
文献类型：
--
作者：
Mahadevan, Jyothi;Jha, Asmita;Rudolph, Johannes;Bowerman, Samuel;Narducci, Domenic;Hansen, Anders S.;Luger, Karolin
PARP 1通过其与染色质的动态关联而有助于基因组结构和DNA损伤修复。PARP 1和PARP 2（PARP 1/2）识别受损的DNA并招募DNA修复机器。在活细胞中使用单分子显微镜，我们监测了PARP 1/2在未受损和受损染色质上的运动。我们确定了两类自由扩散PARP 1/2和两类绑定PARP 1/2。大多数（>60%）的PARP 1/2在未受损和受损的核中以及在用于癌症治疗的PARP 1/2抑制剂（PARPi）的存在下自由扩散。激光诱导的DNA损伤导致一小部分缓慢扩散的PARP 1和PARP 2瞬时结合。在存在DNA损伤的情况下用PARPi处理细胞会导致结合的PARP 1/2的动力学发生微妙变化，但不会引起先前观察到的高水平PARP 1/2捕获。我们的研究结果表明，下一代PARPi可以特异性靶向DNA结合的PARP 1/2的一小部分。活细胞中的单分子显微镜用于监测PARP 1/2细胞用DNA损伤剂和/或PARP 1/2抑制剂处理大多数PARP 1/2自由扩散，而不管DNA损伤或抑制剂的存在，用抑制剂处理仅导致PARP 1/2流动性的细微变化生物科学;细胞生物学;分子生物学;细胞生物学的组织方面
PARP1 contributes to genome architecture and DNA damage repair through its dynamic association with chromatin. PARP1 and PARP2 (PARP1/2) recognize damaged DNA and recruit the DNA repair machinery. Using single-molecule microscopy in live cells, we monitored the movement of PARP1/2 on undamaged and damaged chromatin. We identify two classes of freely diffusing PARP1/2 and two classes of bound PARP1/2. The majority (>60%) of PARP1/2 diffuse freely in both undamaged and damaged nuclei and in the presence of inhibitors of PARP1/2 used for cancer therapy (PARPi). Laser-induced DNA damage results in a small fraction of slowly diffusing PARP1 and PARP2 to become transiently bound. Treatment of cells with PARPi in the presence of DNA damage causes subtle changes in the dynamics of bound PARP1/2, but not the high levels of PARP1/2 trapping seen previously. Our results imply that next-generation PARPi could specifically target the small fraction of DNA-bound PARP1/2. Single-molecule microscopy in live cells was used to monitor PARP1/2 Cells were treated with DNA-damaging agents and/or inhibitors of PARP1/2 Most PARP1/2 diffuse freely regardless of DNA damage or presence of inhibitors Treatment with inhibitors leads to only subtle changes in PARP1/2 mobility Biological sciences; Cell biology; Molecular biology; Organizational aspects of cell biology
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DOI：
10.1038/nrm.2017.53
发表时间：
2017-10
期刊：
Nature reviews. Molecular cell biology
影响因子：
--
作者：
Ray Chaudhuri A;Nussenzweig A
通讯作者：
Nussenzweig A
影响因子：
7.8
作者：
Azad, Gajendra Kumar;Ito, Kenji;Meshorer, Eran
通讯作者：
Meshorer, Eran
影响因子：
16.6
作者：
Caron, Marie-Christine;Sharma, Ajit K.;Masson, Jean-Yves
通讯作者：
Masson, Jean-Yves
影响因子：
11.4
作者：
de Murcia, JMN;Ricoul, M;de Murcia, G
通讯作者：
de Murcia, G
影响因子：
4.8
作者：
Haince, Jean-Francois;McDonald, Darin;Poirier, Guy G.
通讯作者：
Poirier, Guy G.