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Replication Fork Slowing and Reversal upon DNA Damage Require PCNA Polyubiquitination and ZRANB3 DNA Translocase Activity


Vujanovic, Marko; Krietsch, Jana; Raso, Maria Chiara; Terraneo, Nastassja; Zellweger, Ralph; Schmid, Jonas A; Taglialatela, Angelo; Huang, Jen-Wei; Holland, Cory L; Zwicky, Katharina; Herrador, Raquel; Jacobs, Heinz; Cortez, David; Ciccia, Alberto; Penengo, Lorenza; Lopes, Massimo (2017). Replication Fork Slowing and Reversal upon DNA Damage Require PCNA Polyubiquitination and ZRANB3 DNA Translocase Activity. Molecular Cell, 67(5):882-890.e5.

Abstract

DNA damage tolerance during eukaryotic replication is orchestrated by PCNA ubiquitination. While monoubiquitination activates mutagenic translesion synthesis, polyubiquitination activates an error-free pathway, elusive in mammals, enabling damage bypass by template switching. Fork reversal is driven in vitro by multiple enzymes, including the DNA translocase ZRANB3, shown to bind polyubiquitinated PCNA. However, whether this interaction promotes fork remodeling and template switching in vivo was unknown. Here we show that damage-induced fork reversal in mammalian cells requires PCNA ubiquitination, UBC13, and K63-linked polyubiquitin chains, previously involved in error-free damage tolerance. Fork reversal in vivo also requires ZRANB3 translocase activity and its interaction with polyubiquitinated PCNA, pinpointing ZRANB3 as a key effector of error-free DNA damage tolerance. Mutations affecting fork reversal also induced unrestrained fork progression and chromosomal breakage, suggesting fork remodeling as a global fork slowing and protection mechanism. Targeting these fork protection systems represents a promising strategy to potentiate cancer chemotherapy.

Abstract

DNA damage tolerance during eukaryotic replication is orchestrated by PCNA ubiquitination. While monoubiquitination activates mutagenic translesion synthesis, polyubiquitination activates an error-free pathway, elusive in mammals, enabling damage bypass by template switching. Fork reversal is driven in vitro by multiple enzymes, including the DNA translocase ZRANB3, shown to bind polyubiquitinated PCNA. However, whether this interaction promotes fork remodeling and template switching in vivo was unknown. Here we show that damage-induced fork reversal in mammalian cells requires PCNA ubiquitination, UBC13, and K63-linked polyubiquitin chains, previously involved in error-free damage tolerance. Fork reversal in vivo also requires ZRANB3 translocase activity and its interaction with polyubiquitinated PCNA, pinpointing ZRANB3 as a key effector of error-free DNA damage tolerance. Mutations affecting fork reversal also induced unrestrained fork progression and chromosomal breakage, suggesting fork remodeling as a global fork slowing and protection mechanism. Targeting these fork protection systems represents a promising strategy to potentiate cancer chemotherapy.

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Additional indexing

Item Type:Journal Article, refereed, original work
Communities & Collections:04 Faculty of Medicine > Institute of Molecular Cancer Research
07 Faculty of Science > Institute of Molecular Cancer Research
Dewey Decimal Classification:570 Life sciences; biology
610 Medicine & health
Uncontrolled Keywords:Cell Biology, Molecular Biology
Language:English
Date:7 September 2017
Deposited On:21 Sep 2017 12:54
Last Modified:19 Aug 2018 10:18
Publisher:Cell Press (Elsevier)
ISSN:1097-2765
OA Status:Closed
Free access at:Publisher DOI. An embargo period may apply.
Publisher DOI:https://doi.org/10.1016/j.molcel.2017.08.010
PubMed ID:28886337
Project Information:
  • : FunderSNSF
  • : Grant ID31003A_146924
  • : Project TitleMechanistic insights into replication stress, implicated in cancer and neurodegenerative syndromes.
  • : FunderSNSF
  • : Grant ID31003A_166370
  • : Project TitleNon-canonical ubiquitination in the response to DNA damage and DNA replication stress
  • : FunderSNSF
  • : Grant ID31003A_169959
  • : Project TitleReplication fork remodelling upon cancer-relevant genotoxic stress
  • : FunderFP7
  • : Grant ID617102
  • : Project TitleRESTRECA - DNA Replication Stress in Cancer
  • : FunderSNSF
  • : Grant ID31003A_146924
  • : Project TitleMechanistic insights into replication stress, implicated in cancer and neurodegenerative syndromes.
  • : FunderSNSF
  • : Grant ID31003A_166370
  • : Project TitleNon-canonical ubiquitination in the response to DNA damage and DNA replication stress
  • : FunderSNSF
  • : Grant ID31003A_169959
  • : Project TitleReplication fork remodelling upon cancer-relevant genotoxic stress

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