ATR-Mediated Global Fork Slowing and Reversal Assist Fork Traverse and Prevent Chromosomal Breakage at DNA Interstrand Cross-Links

Karun Mutreja, Jana Krietsch, Jeannine Hess, Sebastian Ursich, Matteo Berti, Fabienne K. Roessler, Ralph Zellweger, Malay Patra, Gilles Gasser, Massimo Lopes*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review


Interstrand cross-links (ICLs) are toxic DNA lesions interfering with DNA metabolism that are induced by widely used anticancer drugs. They have long been considered absolute roadblocks for replication forks, implicating complex DNA repair processes at stalled or converging replication forks. Recent evidence challenged this view, proposing that single forks traverse ICLs by yet elusive mechanisms. Combining ICL immunolabeling and single-molecule approaches in human cells, we now show that ICL induction leads to global replication fork slowing, involving forks not directly challenged by ICLs. Active fork slowing is linked to rapid recruitment of RAD51 to replicating chromatin and to RAD51/ZRANB3-mediated fork reversal. This global modulation of fork speed and architecture requires ATR activation, promotes single-fork ICL traverse—here, directly visualized by electron microscopy—and prevents chromosomal breakage by untimely ICL processing. We propose that global fork slowing by remodeling provides more time for template repair and promotes bypass of residual lesions, limiting fork-associated processing. Replication-coupled repair of DNA interstrand cross-links (ICLs) promotes resistance to chemotherapeutic treatments. Visualizing individual lesions and replication intermediates, Mutreja et al. report that forks slow down and reverse both at ICLs and away from lesions. This ATR-mediated response assists lesion bypass during replication and limits chromosomal breakage by fork-associated processing.

Original languageEnglish
Pages (from-to)2629-2642.e5
JournalCell Reports
Issue number10
StatePublished - 4 Sep 2018
Externally publishedYes


  • ATR checkpoint
  • DNA interstrand crosslinks
  • DNA replication
  • DNA replication stress response
  • ICL immunolabeling
  • electron microscopy
  • fork traverse
  • global fork slowing
  • replication fork reversal


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