RPA-Mediated Recruitment of the E3 Ligase RFWD3 Is Vital for Interstrand Crosslink Repair and Human Health

Laura Feeney, Ivan M. Munoz, Christophe Lachaud, Rachel Toth, Paul L. Appleton, Detlev Schindler, John Rouse (Lead / Corresponding author)

Research output: Contribution to journalArticle

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Abstract

Defects in the repair of DNA inter-strand crosslinks (ICLs) are associated with the genome instability syndrome Fanconi anemia (FA). Here we report that cells with mutations in RFWD3, an E3 ubiquitin ligase that interacts with and ubiquitylates replication protein A (RPA), show profound defects in ICL repair. An amino acid substitution in the WD40 repeats of RFWD3 (I639K) found in a new FA subtype abolishes interaction of RFWD3 with RPA, thereby preventing RFWD3 recruitment to sites of ICL-induced replication fork stalling. Moreover, single point mutations in the RPA32 subunit of RPA that abolish interaction with RWFD3 also inhibit ICL repair, demonstrating that RPA-mediated RFWD3 recruitment to stalled replication forks is important for ICL repair. We also report that unloading of RPA from sites of ICL induction is perturbed in RFWD3-deficient cells. These data reveal important roles for RFWD3 localization in protecting genome stability and preserving human health.
Original languageEnglish
Pages (from-to)610-621.e4
Number of pages16
JournalMolecular Cell
Volume66
Issue number5
Early online date1 Jun 2017
DOIs
Publication statusPublished - 1 Jun 2017

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Replication Protein A
Ubiquitin-Protein Ligases
Health
Fanconi Anemia
Genomic Instability
Amino Acid Substitution
Point Mutation
DNA Repair
Mutation

Keywords

  • RFWD3
  • ICL
  • Fanconi anemia
  • RPA
  • Homologous recombination
  • Ubiquitylation

Cite this

@article{3894fe5cdfbd4fabb136516d5ac4379f,
title = "RPA-Mediated Recruitment of the E3 Ligase RFWD3 Is Vital for Interstrand Crosslink Repair and Human Health",
abstract = "Defects in the repair of DNA inter-strand crosslinks (ICLs) are associated with the genome instability syndrome Fanconi anemia (FA). Here we report that cells with mutations in RFWD3, an E3 ubiquitin ligase that interacts with and ubiquitylates replication protein A (RPA), show profound defects in ICL repair. An amino acid substitution in the WD40 repeats of RFWD3 (I639K) found in a new FA subtype abolishes interaction of RFWD3 with RPA, thereby preventing RFWD3 recruitment to sites of ICL-induced replication fork stalling. Moreover, single point mutations in the RPA32 subunit of RPA that abolish interaction with RWFD3 also inhibit ICL repair, demonstrating that RPA-mediated RFWD3 recruitment to stalled replication forks is important for ICL repair. We also report that unloading of RPA from sites of ICL induction is perturbed in RFWD3-deficient cells. These data reveal important roles for RFWD3 localization in protecting genome stability and preserving human health.",
keywords = "RFWD3, ICL, Fanconi anemia, RPA, Homologous recombination, Ubiquitylation",
author = "Laura Feeney and Munoz, {Ivan M.} and Christophe Lachaud and Rachel Toth and Appleton, {Paul L.} and Detlev Schindler and John Rouse",
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RPA-Mediated Recruitment of the E3 Ligase RFWD3 Is Vital for Interstrand Crosslink Repair and Human Health. / Feeney, Laura; Munoz, Ivan M.; Lachaud, Christophe; Toth, Rachel; Appleton, Paul L.; Schindler, Detlev; Rouse, John (Lead / Corresponding author).

In: Molecular Cell, Vol. 66, No. 5, 01.06.2017, p. 610-621.e4.

Research output: Contribution to journalArticle

TY - JOUR

T1 - RPA-Mediated Recruitment of the E3 Ligase RFWD3 Is Vital for Interstrand Crosslink Repair and Human Health

AU - Feeney, Laura

AU - Munoz, Ivan M.

AU - Lachaud, Christophe

AU - Toth, Rachel

AU - Appleton, Paul L.

AU - Schindler, Detlev

AU - Rouse, John

N1 - Funding: MRC (Grant: MC_UU_12016/1)

PY - 2017/6/1

Y1 - 2017/6/1

N2 - Defects in the repair of DNA inter-strand crosslinks (ICLs) are associated with the genome instability syndrome Fanconi anemia (FA). Here we report that cells with mutations in RFWD3, an E3 ubiquitin ligase that interacts with and ubiquitylates replication protein A (RPA), show profound defects in ICL repair. An amino acid substitution in the WD40 repeats of RFWD3 (I639K) found in a new FA subtype abolishes interaction of RFWD3 with RPA, thereby preventing RFWD3 recruitment to sites of ICL-induced replication fork stalling. Moreover, single point mutations in the RPA32 subunit of RPA that abolish interaction with RWFD3 also inhibit ICL repair, demonstrating that RPA-mediated RFWD3 recruitment to stalled replication forks is important for ICL repair. We also report that unloading of RPA from sites of ICL induction is perturbed in RFWD3-deficient cells. These data reveal important roles for RFWD3 localization in protecting genome stability and preserving human health.

AB - Defects in the repair of DNA inter-strand crosslinks (ICLs) are associated with the genome instability syndrome Fanconi anemia (FA). Here we report that cells with mutations in RFWD3, an E3 ubiquitin ligase that interacts with and ubiquitylates replication protein A (RPA), show profound defects in ICL repair. An amino acid substitution in the WD40 repeats of RFWD3 (I639K) found in a new FA subtype abolishes interaction of RFWD3 with RPA, thereby preventing RFWD3 recruitment to sites of ICL-induced replication fork stalling. Moreover, single point mutations in the RPA32 subunit of RPA that abolish interaction with RWFD3 also inhibit ICL repair, demonstrating that RPA-mediated RFWD3 recruitment to stalled replication forks is important for ICL repair. We also report that unloading of RPA from sites of ICL induction is perturbed in RFWD3-deficient cells. These data reveal important roles for RFWD3 localization in protecting genome stability and preserving human health.

KW - RFWD3

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DO - 10.1016/j.molcel.2017.04.021

M3 - Article

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