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Disruption of mouse Slx4, a regulator of structure-specific nucleases, phenocopies Fanconi anemia

Nature Genetics volume 43, pages 147–152 (2011)Cite this article

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Abstract

The evolutionarily conserved SLX4 protein, a key regulator of nucleases, is critical for DNA damage response. SLX4 nuclease complexes mediate repair during replication and can also resolve Holliday junctions formed during homologous recombination. Here we describe the phenotype of the Btbd12 knockout mouse, the mouse ortholog of SLX4, which recapitulates many key features of the human genetic illness Fanconi anemia. Btbd12-deficient animals are born at sub-Mendelian ratios, have greatly reduced fertility, are developmentally compromised and are prone to blood cytopenias. Btbd12−/− cells prematurely senesce, spontaneously accumulate damaged chromosomes and are particularly sensitive to DNA crosslinking agents. Genetic complementation reveals a crucial requirement for Btbd12 (also known as Slx4) to interact with the structure-specific endonuclease Xpf-Ercc1 to promote crosslink repair. The Btbd12 knockout mouse therefore establishes a disease model for Fanconi anemia and genetically links a regulator of nuclease incision complexes to the Fanconi anemia DNA crosslink repair pathway.

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Figure 1: Btbd12 deficiency results in growth retardation and compromised fertility.
Figure 2: Some Btbd12−/− mice died prematurely and displayed brain and eye developmental defects.
Figure 3: A proportion of Btbd12−/− mice developed blood cytopenias associated with genomic instability.
Figure 4: Btbd12-deficient cells undergo premature replicative senescence, exhibiting spontaneous and inducible chromosomal instability.
Figure 5: Btbd12-deficient MEFs are hypersensitive to DNA interstrand crosslinking agents.
Figure 6: The interaction between Slx4 and Xpf-Ercc1 is required for crosslink repair.

References

  1. Ciccia, A., McDonald, N. & West, S.C. Structural and functional relationships of the XPF/MUS81 family of proteins. Annu. Rev. Biochem. 77, 259–287 (2008).

    Article  CAS  Google Scholar 

  2. Svendsen, J.M. & Harper, J.W. GEN1/Yen1 and the SLX4 complex: Solutions to the problem of Holliday junction resolution. Genes Dev. 24, 521–536 (2010).

    Article  CAS  Google Scholar 

  3. Coulon, S. et al. Slx1-Slx4 are subunits of a structure-specific endonuclease that maintains ribosomal DNA in fission yeast. Mol. Biol. Cell 15, 71–80 (2004).

    Article  CAS  Google Scholar 

  4. Fricke, W.M. & Brill, S.J. Slx1-Slx4 is a second structure-specific endonuclease functionally redundant with Sgs1-Top3. Genes Dev. 17, 1768–1778 (2003).

    Article  CAS  Google Scholar 

  5. Fekairi, S. et al. Human SLX4 is a Holliday junction resolvase subunit that binds multiple DNA repair/recombination endonucleases. Cell 138, 78–89 (2009).

    Article  CAS  Google Scholar 

  6. Svendsen, J.M. et al. Mammalian BTBD12/SLX4 assembles a Holliday junction resolvase and is required for DNA repair. Cell 138, 63–77 (2009).

    Article  CAS  Google Scholar 

  7. Munõz, I.M. et al. Coordination of structure-specific nucleases by human SLX4/BTBD12 is required for DNA repair. Mol. Cell 35, 116–127 (2009).

    Article  Google Scholar 

  8. Flott, S. et al. Phosphorylation of Slx4 by Mec1 and Tel1 regulates the single-strand annealing mode of DNA repair in budding yeast. Mol. Cell. Biol. 27, 6433–6445 (2007).

    Article  CAS  Google Scholar 

  9. Matsuoka, S. et al. ATM and ATR substrate analysis reveals extensive protein networks responsive to DNA damage. Science 316, 1160–1166 (2007).

    Article  CAS  Google Scholar 

  10. Cheng, N.C. et al. Mice with a targeted disruption of the Fanconi anemia homolog Fanca. Hum. Mol. Genet. 9, 1805–1811 (2000).

    Article  CAS  Google Scholar 

  11. Lu, B. & Bishop, C.E. Late onset of spermatogenesis and gain of fertility in POG-deficient mice indicate that POG is not necessary for the proliferation of spermatogonia. Biol. Reprod. 69, 161–168 (2003).

    Article  CAS  Google Scholar 

  12. Agoulnik, A.I. et al. A novel gene, Pog, is necessary for primordial germ cell proliferation in the mouse and underlies the germ cell deficient mutation, gcd. Hum. Mol. Genet. 11, 3047–3053 (2002).

    Article  CAS  Google Scholar 

  13. Houghtaling, S. et al. Epithelial cancer in Fanconi anemia complementation group D2 (Fancd2) knockout mice. Genes Dev. 17, 2021–2035 (2003).

    Article  CAS  Google Scholar 

  14. Adams, D.J. et al. BRCTx is a novel, highly conserved RAD18-interacting protein. Mol. Cell. Biol. 25, 779–788 (2005).

    Article  CAS  Google Scholar 

  15. Patel, K.J. & Joenje, H. Fanconi anemia and DNA replication repair. DNA Repair (Amst.) 6, 885–890 (2007).

    Article  CAS  Google Scholar 

  16. Zhang, X.Y. et al. Xpf and not the Fanconi anaemia proteins or Rev3 accounts for the extreme resistance to cisplatin in Dictyostelium discoideum. PLoS Genet. 5, e1000645 (2009).

    Article  Google Scholar 

  17. Dendouga, N. et al. Disruption of murine Mus81 increases genomic instability and DNA damage sensitivity but does not promote tumorigenesis. Mol. Cell. Biol. 25, 7569–7579 (2005).

    Article  CAS  Google Scholar 

  18. Gillio, A.P., Verlander, P.C., Batish, S.D., Giampietro, P.F. & Auerbach, A.D. Phenotypic consequences of mutations in the Fanconi anemia FAC gene: an International Fanconi Anemia Registry study. Blood 90, 105–110 (1997).

    CAS  PubMed  Google Scholar 

  19. Hsia, K.T. et al. DNA repair gene Ercc1 is essential for normal spermatogenesis and oogenesis and for functional integrity of germ cell DNA in the mouse. Development 130, 369–378 (2003).

    Article  CAS  Google Scholar 

  20. Weeda, G. et al. Disruption of mouse Ercc1 results in a novel repair syndrome with growth failure, nuclear abnormalities and senescence. Curr. Biol. 7, 427–439 (1997).

    Article  CAS  Google Scholar 

  21. McWhir, J., Selfridge, J., Harrison, D.J., Squires, S. & Melton, D.W. Mice with DNA repair gene (Ercc1) deficiency have elevated levels of p53, liver nuclear abnormalities and die before weaning. Nat. Genet. 5, 217–224 (1993).

    Article  CAS  Google Scholar 

  22. Selfridge, J., Hsia, K.T., Redhead, N.J. & Melton, D.W. Correction of liver dysfunction in DNA repair-deficient mice with an ERCC1 transgene. Nucleic Acids Res. 29, 4541–4550 (2001).

    Article  CAS  Google Scholar 

  23. Lawrence, N.J., Sacco, J.J., Brownstein, D.G., Gillingwater, T.H. & Melton, D.W. A neurological phenotype in mice with DNA repair gene Ercc1 deficiency. DNA Repair (Amst.) 7, 281–291 (2008).

    Article  CAS  Google Scholar 

  24. de Waard, M.C. et al. Age-related motor neuron degeneration in DNA repair-deficient Ercc1 mice. Acta Neuropathol. 120, 461–475 (2010).

    Article  Google Scholar 

  25. Stoepker, C. et al. SLX4, a coordinator of structure-specific endonucleases, is mutated in a new Fanconi anemia subtype. Nat. Genet. advance online publication, doi:10.1038/ng.751 (16 January 2011).

    Article  CAS  Google Scholar 

  26. Pace, P. et al. FANCE: the link between Fanconi anaemia complex assembly and activity. EMBO J. 21, 3414–3423 (2002).

    Article  CAS  Google Scholar 

  27. Hancock, J.M. et al. Mouse Phenotype Database Integration Consortium: integration [corrected] of mouse phenome data resources. Mamm. Genome 18, 157–163 (2007).

    Article  Google Scholar 

  28. Masuya, H. et al. Implementation of the modified-SHIRPA protocol for screening of dominant phenotypes in a large-scale ENU mutagenesis program. Mamm. Genome 16, 829–837 (2005).

    Article  Google Scholar 

Download references

Acknowledgements

We wish to thank L. Neidernhofer for the kind gift of Ercc1-deficient and wild-type congenic MEFs. Fanca- and Fancc-deficient MEFs were obtained from the Fanconi Anaemia Research Fund Cell Repository, Eugene, Oregon, USA. We thank A. Shortland for substantial contributions in the planning of animal experiments. The authors would like to thank M. Taylor for cloning of the mouse Slx4 cDNA. We would also like to thank T. Langford, J. Cruickshank, A. Mead, M. Brown, J. Wiles, K. Robinson, C. Shepherd, M. Reed, V. Smith and R. Berks for assistance in animal husbandry and handling. We would also like to thank G. Nemeth (Mediso Ltd.) for assistance with the computed tomography methodology. D.J.A. was supported by Cancer Research-UK and the Wellcome Trust. P.H.L.G. is supported by ATIP-Centre National de la Recherche Scientifique, Association pour la Recherche sur le Cancer and Fondation pour la Recherche Medicale grants.

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Authors and Affiliations

  1. Medical Research Council, Laboratory of Molecular Biology, Cambridge, UK

    Gerry P Crossan, Ivan V Rosado, Frederic Langevin & Ketan J Patel

  2. Wellcome Trust Sanger Institute, Wellcome Trust Genome Campus, Cambridge, UK

    Louise van der Weyden, Rebecca E McIntyre & David J Adams

  3. Genome Instability and Carcinogenesis UPR3081 Centre National de la Recherche Scientifique (CNRS), Conventionné par l'Université d'Aix-Marseille 2, Marseille, France

    Pierre-Henri L Gaillard

  4. Cancer Research UK, Cambridge Research Institute, Li Ka Shing Centre, Cambridge, UK

    Ferdia Gallagher, Mikko I Kettunen, David Y Lewis & Kevin Brindle

  5. Department of Pathology, Addenbrooke's Hospital, University of Cambridge, Cambridge, UK

    Mark J Arends

  6. Department of Medicine, Addenbrooke's Hospital, University of Cambridge, Cambridge, UK

    Ketan J Patel

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  1. Gerry P Crossan
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Sanger Mouse Genetics Project

Contributions

K.J.P. and G.P.C. designed the study, the experiments and wrote the paper. G.P.C. performed the majority of the experiments presented. D.J.A. helped in the design of the experiments. M.J.A. analyzed histological samples and provided useful discussion. L.v.d.W. managed the colony and weighed and performed necropsies on mice at Wellcome Trust Sanger Institute. I.V.R. analyzed Fanconi pathway activation. F.L. assisted in analysis of developmental abnormalities. P.-H.L.G. helped in the design of some experiments. R.E.M. performed the micronucleus assay. S.M.G.P. performed the mouse phenotype pipeline analysis. F.G., M.I.K., D.Y.L. and K.B. performed imaging studies.

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Correspondence to Ketan J Patel.

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Crossan, G., van der Weyden, L., Rosado, I. et al. Disruption of mouse Slx4, a regulator of structure-specific nucleases, phenocopies Fanconi anemia. Nat Genet 43, 147–152 (2011). https://doi.org/10.1038/ng.752

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