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Cell Mol Life Sci. 2019 Aug 7. doi: 10.1007/s00018-019-03264-5. [Epub ahead of print]

ERCC1-XPF targeting to psoralen-DNA crosslinks depends on XPA and FANCD2.

Author information

1
Department of Molecular Genetics, Erasmus MC, 3015 GE, Rotterdam, The Netherlands.
2
Oncode Institute, Erasmus MC, 3015 GE, Rotterdam, The Netherlands.
3
CeMM Research Centre for Molecular Medicine of the Austrian Academy of Sciences, 1090, Vienna, Austria.
4
Department of Molecular Genetics, Erasmus MC, 3015 GE, Rotterdam, The Netherlands. w.vermeulen@erasmusmc.nl.
5
Oncode Institute, Erasmus MC, 3015 GE, Rotterdam, The Netherlands. w.vermeulen@erasmusmc.nl.
6
Department of Molecular Genetics, Erasmus MC, 3015 GE, Rotterdam, The Netherlands. w.lans@erasmusmc.nl.
7
Oncode Institute, Erasmus MC, 3015 GE, Rotterdam, The Netherlands. w.lans@erasmusmc.nl.

Abstract

The effectiveness of many DNA-damaging chemotherapeutic drugs depends on their ability to form monoadducts, intrastrand crosslinks and/or interstrand crosslinks (ICLs) that interfere with transcription and replication. The ERCC1-XPF endonuclease plays a critical role in removal of these lesions by incising DNA either as part of nucleotide excision repair (NER) or interstrand crosslink repair (ICLR). Engagement of ERCC1-XPF in NER is well characterized and is facilitated by binding to the XPA protein. However, ERCC1-XPF recruitment to ICLs is less well understood. Moreover, specific mutations in XPF have been found to disrupt its function in ICLR but not in NER, but whether this involves differences in lesion targeting is unknown. Here, we imaged GFP-tagged ERCC1, XPF and ICLR-defective XPF mutants to investigate how in human cells ERCC1-XPF is localized to different types of psoralen-induced DNA lesions, repaired by either NER or ICLR. Our results confirm its dependence on XPA in NER and furthermore show that its engagement in ICLR is dependent on FANCD2. Interestingly, we find that two ICLR-defective XPF mutants (R689S and S786F) are less well recruited to ICLs. These studies highlight the differential mechanisms that regulate ERCC1-XPF activity in DNA repair.

KEYWORDS:

DNA damage response; DNA repair; Fanconi anemia; Xeroderma pigmentosum–Cockayne syndrome complex

PMID:
31392348
DOI:
10.1007/s00018-019-03264-5

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