Recruitment and positioning determine the specific role of the XPF-ERCC1 endonuclease in interstrand crosslink repair

Daisy Klein Douwel, Wouter S Hoogenboom, Rick Acm Boonen, Puck Knipscheer

Research output: Contribution to journal/periodicalArticleScientificpeer-review

34 Citations (Scopus)

Abstract

XPF-ERCC1 is a structure-specific endonuclease pivotal for several DNA repair pathways and, when mutated, can cause multiple diseases. Although the disease-specific mutations are thought to affect different DNA repair pathways, the molecular basis for this is unknown. Here we examine the function of XPF-ERCC1 in DNA interstrand crosslink (ICL) repair. We used Xenopus egg extracts to measure both ICL and nucleotide excision repair, and we identified mutations that are specifically defective in ICL repair. One of these separation-of-function mutations resides in the helicase-like domain of XPF and disrupts binding to SLX4 and recruitment to the ICL A small deletion in the same domain supports recruitment of XPF to the ICL, but inhibited the unhooking incisions most likely by disrupting a second, transient interaction with SLX4. Finally, mutation of residues in the nuclease domain did not affect localization of XPF-ERCC1 to the ICL but did prevent incisions on the ICL substrate. Our data support a model in which the ICL repair-specific function of XPF-ERCC1 is dependent on recruitment, positioning and substrate recognition.

Original languageEnglish
Pages (from-to)2034-2046
Number of pages13
JournalEMBO Journal
Volume36
Issue number14
DOIs
Publication statusPublished - 14 Jul 2017

Keywords

  • Animals
  • DNA Repair
  • DNA Repair Enzymes
  • Models, Biological
  • Mutant Proteins
  • Mutation
  • Xenopus
  • Journal Article
  • Research Support, Non-U.S. Gov't

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