Differential binding kinetics of replication protein A during replication and the pre- and post-incision steps of nucleotide excision repair

Audrey Gourdin, Loes Cuijk, Maria Tresini, MS Luijsterburg, Alex Nigg, GC Giglia, Adriaan Houtsmuller, Wim Vermeulen, Jurgen Marteijn

Research output: Contribution to journalArticleAcademic

1 Citation (Scopus)

Abstract

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.
Original languageUndefined/Unknown
Pages (from-to)46-56
Number of pages11
JournalDNA Repair
Volume24
DOIs
Publication statusPublished - 2014

Cite this