Journal article
Identification of KIAA1018/FAN1, a DNA Repair Nuclease Recruited to DNA Damage by Monoubiquitinated FANCD2
C MacKay, AC Déclais, C Lundin, A Agostinho, AJ Deans, TJ MacArtney, K Hofmann, A Gartner, SC West, T Helleday, DMJ Lilley, J Rouse
Cell | Published : 2010
Abstract
DNA interstrand crosslinks (ICLs) are highly toxic because they block the progression of replisomes. The Fanconi Anemia (FA) proteins, encoded by genes that are mutated in FA, are important for repair of ICLs. The FA core complex catalyzes the monoubiquitination of FANCD2, and this event is essential for several steps of ICL repair. However, how monoubiquitination of FANCD2 promotes ICL repair at the molecular level is unknown. Here, we describe a highly conserved protein, KIAA1018/MTMR15/FAN1, that interacts with, and is recruited to sites of DNA damage by, the monoubiquitinated form of FANCD2. FAN1 exhibits endonuclease activity toward 5' flaps and has 5' exonuclease activity, and these ac..
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Awarded by Fanconi Anemia Research Fund
Funding Acknowledgements
We are grateful to Katja Kratz and Joe Jiricny for experimental advice, for helping us with the purification of FAN1, and for communicating results prior to publication. We thank K.J. Patel, Johan de Winter, and Maureen Hoatlin for kind gifts of antibodies. pDEST40-V5-FANCD2 wild-type and pDEST40-V5-FANCD2 K561R plasmids were kind gifts from Niall Howlett and Thomas Glover. We thank Alan d'Andrea for the kind gifts of FANCD2<SUP>-/-</SUP> fibroblasts (PD20 cells) corrected with vector, FANCD2 wild-type, or FANCD2 K561R. We are grateful to James Hastie, Hilary MacLauchlan, and the Antibody Production Team at Division of Signal Transduction Therapy, University of Dundee, to Bob Gourlay, Sanjay Kothiya, and Nick Morrice for help with mass spectrometry, and to the DNA Sequencing Service at the College of Life Sciences (CLS), University of Dundee. We are grateful to the microscopy facility, CLS, University of Dundee for assistance with microscopy. We thank Jim Haber and the J.R. lab for critical reading of the manuscript. This work was funded by the UK Medical Research Council (C.M., T.J.H., and J.R.), Cancer Research UK (A.C.D, A.G., and D.M.J.L.), and the Fanconi Anemia Research Fund (A.J.D. and S.C.W.).