MRE11-RAD50-NBS1 is a critical regulator of FANCD2 stability and function during DNA double-strand break repair

Céline Roques; Yan Coulombe; Mathieu Delannoy; Julien Vignard; Simona Grossi; Isabelle Brodeur; Amélie Rodrigue; Jean Gautier; Alicja Z Stasiak; Andrzej Stasiak; Angelos Constantinou; Jean-Yves Masson

doi:10.1038/emboj.2009.193

MRE11-RAD50-NBS1 is a critical regulator of FANCD2 stability and function during DNA double-strand break repair

EMBO J. 2009 Aug 19;28(16):2400-13. doi: 10.1038/emboj.2009.193. Epub 2009 Jul 16.

Authors

Céline Roques¹, Yan Coulombe, Mathieu Delannoy, Julien Vignard, Simona Grossi, Isabelle Brodeur, Amélie Rodrigue, Jean Gautier, Alicja Z Stasiak, Andrzej Stasiak, Angelos Constantinou, Jean-Yves Masson

Affiliation

¹ Genome Stability Laboratory, Laval University Cancer Research Center, Hôtel-Dieu de Québec, Québec, Canada.

Abstract

Monoubiquitination of the Fanconi anaemia protein FANCD2 is a key event leading to repair of interstrand cross-links. It was reported earlier that FANCD2 co-localizes with NBS1. However, the functional connection between FANCD2 and MRE11 is poorly understood. In this study, we show that inhibition of MRE11, NBS1 or RAD50 leads to a destabilization of FANCD2. FANCD2 accumulated from mid-S to G2 phase within sites containing single-stranded DNA (ssDNA) intermediates, or at sites of DNA damage, such as those created by restriction endonucleases and laser irradiation. Purified FANCD2, a ring-like particle by electron microscopy, preferentially bound ssDNA over various DNA substrates. Inhibition of MRE11 nuclease activity by Mirin decreased the number of FANCD2 foci formed in vivo. We propose that FANCD2 binds to ssDNA arising from MRE11-processed DNA double-strand breaks. Our data establish MRN as a crucial regulator of FANCD2 stability and function in the DNA damage response.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Acid Anhydride Hydrolases
Cell Cycle Proteins / genetics
Cell Cycle Proteins / metabolism*
DNA Breaks, Double-Stranded*
DNA Repair Enzymes / genetics
DNA Repair Enzymes / metabolism*
DNA Repair*
DNA-Binding Proteins / genetics
DNA-Binding Proteins / metabolism*
Down-Regulation
Fanconi Anemia Complementation Group D2 Protein / analysis
Fanconi Anemia Complementation Group D2 Protein / genetics
Fanconi Anemia Complementation Group D2 Protein / metabolism*
HeLa Cells
Humans
MRE11 Homologue Protein
Microscopy, Electron
Nuclear Proteins / genetics
Nuclear Proteins / metabolism*
Protein Binding
Protein Stability
RNA, Small Interfering / genetics

Substances

Cell Cycle Proteins
DNA-Binding Proteins
FANCD2 protein, human
Fanconi Anemia Complementation Group D2 Protein
MRE11 protein, human
NBN protein, human
Nuclear Proteins
RNA, Small Interfering
MRE11 Homologue Protein
Acid Anhydride Hydrolases
RAD50 protein, human
DNA Repair Enzymes