MRE11–RAD50–NBS1 is a critical regulator of FANCD2 stability and function during DNA double‐strand break repair |
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| Authors: | 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 |
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| Affiliation: | 1. Genome Stability Laboratory, Laval University Cancer Research Center, H?tel‐Dieu de Québec, Québec, Canada;2. Department of Biochemistry, University of Lausanne, BIL Biomedical Research Center, Lausanne, Switzerland;3. Columbia University, Institute for Cancer Genetics, Irving Cancer Research Center, New York, NY, USA;4. Center for Integrative Genomics, Faculty of Biology and Medicine, University of Lausanne, Lausanne, Switzerland |
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| 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. |
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| Keywords: | DNA metabolism Fanconi anaemia homologous recombination Mirin |
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