Subnuclear Relocalization of Structure-Specific Endonucleases in Response to DNA Damage

Cell Rep. 2017 Aug 15;20(7):1553-1562. doi: 10.1016/j.celrep.2017.07.059.

Abstract

Structure-specific endonucleases contribute to the maintenance of genome integrity by cleaving DNA intermediates that need to be resolved for faithful DNA repair, replication, or recombination. Despite advances in the understanding of their function and regulation, it is less clear how these proteins respond to genotoxic stress. Here, we show that the structure-specific endonuclease Mus81-Mms4/EME1 relocalizes to subnuclear foci following DNA damage and colocalizes with the endonucleases Rad1-Rad10 (XPF-ERCC1) and Slx1-Slx4. Recruitment takes place into a class of stress foci defined by Cmr1/WDR76, a protein involved in preserving genome stability, and depends on the E2-ubiquitin-conjugating enzyme Rad6 and the E3-ubiquitin ligase Bre1. Foci dynamics show that, in the presence of DNA intermediates that need resolution by Mus81-Mms4, Mus81 foci persist until this endonuclease is activated by Mms4 phosphorylation. Our data suggest that subnuclear relocalization is relevant for the function of Mus81-Mms4 and, probably, of the endonucleases that colocalize with it.

Keywords: DNA damage response; Mus81-Mms4; protein relocalization; structure-specific endonucleases; subnuclear foci.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • DNA Damage
  • DNA Repair Enzymes / genetics
  • DNA Repair Enzymes / metabolism
  • DNA Repair*
  • DNA Replication
  • DNA, Fungal / genetics*
  • DNA, Fungal / metabolism
  • DNA-Binding Proteins / genetics*
  • DNA-Binding Proteins / metabolism
  • Endodeoxyribonucleases / genetics
  • Endodeoxyribonucleases / metabolism
  • Endonucleases / genetics*
  • Endonucleases / metabolism
  • Flap Endonucleases / genetics*
  • Flap Endonucleases / metabolism
  • Gene Expression Regulation, Fungal*
  • Phosphorylation
  • Protein Binding
  • Protein Transport
  • Saccharomyces cerevisiae / genetics*
  • Saccharomyces cerevisiae / metabolism
  • Saccharomyces cerevisiae Proteins / genetics*
  • Saccharomyces cerevisiae Proteins / metabolism
  • Signal Transduction
  • Single-Strand Specific DNA and RNA Endonucleases / genetics
  • Single-Strand Specific DNA and RNA Endonucleases / metabolism
  • Ubiquitin-Conjugating Enzymes / genetics
  • Ubiquitin-Conjugating Enzymes / metabolism

Substances

  • Bre1 protein, S cerevisiae
  • Cmr1 protein, S cerevisiae
  • DNA, Fungal
  • DNA-Binding Proteins
  • Saccharomyces cerevisiae Proteins
  • RAD6 protein, S cerevisiae
  • Ubiquitin-Conjugating Enzymes
  • Endodeoxyribonucleases
  • Endonucleases
  • Flap Endonucleases
  • MUS81 protein, S cerevisiae
  • RAD1 protein, S cerevisiae
  • SLX1 protein, S cerevisiae
  • SLX4 protein, S cerevisiae
  • MMS4 protein, S cerevisiae
  • RAD10 protein, S cerevisiae
  • Single-Strand Specific DNA and RNA Endonucleases
  • DNA Repair Enzymes