Differential binding kinetics of replication protein A during replication and the pre- and post-incision steps of nucleotide excision repair
Differential binding kinetics of replication protein A during replication and the pre- and post-incision steps of nucleotide excision repair
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DOI：
10.1016/j.dnarep.2014.09.013
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发表时间：
2014-12-01
期刊：
影响因子：
3.8
通讯作者：
Marteijn, Jurgen A.
中科院分区：
文献类型：
--
作者：
Gourdin, Audrey M.;van Cuijk, Loes;Marteijn, Jurgen A.
复制蛋白A（RPA）结合单链DNA（ssDNA）的能力强调了其在DNA复制和修复过程中的关键作用。GFP标记的RPA 70的免疫荧光和活细胞成像的组合显示，RPA，与其他复制因子相比，不聚集成复制焦点，这是由其在ssDNA的短停留时间解释的。除了复制之外，RPA还在核苷酸切除修复（NER）的切割前和切割后步骤中起着至关重要的作用。切口前因子如XPC和TFIIH在局部诱导的UV损伤中迅速积累，并在4小时内保持可见。然而，RPA没有达到其最大的积累水平，直到3小时后，DNA损伤的施加和染色质结合池仍然可检测到8小时，可能反映了它的作用，在后切口步骤的NER。在NER的预切割步骤期间，RPA只能在切口缺陷XP-F细胞中的DNA损伤处可见，然而在DNA损伤处的停留时间没有实质性增加。总之，我们的数据表明，RPA是一个内在的高度动态的ssDNA结合复合物在复制和NER的不同步骤。(C)2014爱思唯尔有限公司版权所有。
The ability of replication protein A (RPA) to bind single-stranded DNA (ssDNA) underlines its crucial roles during DNA replication and repair. A combination of immunofluorescence and live cell imaging of GFP-tagged RPA70 revealed that RPA, in contrast to other replication factors, does not cluster into replication foci, which is explained by its short residence time at ssDNA. In addition to replication, RPA also plays a crucial role in both the pre- and post-incision steps of nucleotide excision repair (NER). Pre-incision factors like XPC and TFIIH accumulate rapidly at locally induced UV-damage and remain visible up to 4h. However, RPA did not reach its maximum accumulation level until 3 h after DNA damage infliction and a chromatin-bound pool remained detectable up to 8 h, probably reflecting its role during the post-incision step of NER. During the pre-incision steps of NER, RPA could only be visualized at DNA lesions in incision deficient XP-F cells, however without a substantial increase in residence time at DNA damage. Together our data show that RPA is an intrinsically highly dynamic ssDNA-binding complex during both replication and distinct steps of NER. (C) 2014 Elsevier B.V. All rights reserved.