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FANCG promotes formation of a newly identified protein complex containing BRCA2, FANCD2 and XRCC3

Oncogene volume 27, pages 3641–3652 (2008)Cite this article

Abstract

Fanconi anemia (FA) is a human disorder characterized by cancer susceptibility and cellular sensitivity to DNA crosslinks and other damages. Thirteen complementation groups and genes are identified, including BRCA2, which is defective in the FA-D1 group. Eight of the FA proteins, including FANCG, participate in a nuclear core complex that is required for the monoubiquitylation of FANCD2 and FANCI. FANCD2, like FANCD1/BRCA2, is not part of the core complex, and we previously showed direct BRCA2–FANCD2 interaction using yeast two-hybrid analysis. We now show in human and hamster cells that expression of FANCG protein, but not the other core complex proteins, is required for co-precipitation of BRCA2 and FANCD2. We also show that phosphorylation of FANCG serine 7 is required for its co-precipitation with BRCA2, XRCC3 and FANCD2, as well as the direct interaction of BRCA2–FANCD2. These results argue that FANCG has a role independent of the FA core complex, and we propose that phosphorylation of serine 7 is the signalling event required for forming a discrete complex comprising FANCD1/BRCA2-FANCD2-FANCG-XRCC3 (D1-D2-G-X3). Cells that fail to express either phospho-Ser7-FANCG, or full length BRCA2 protein, lack the interactions amongst the four component proteins. A role for D1-D2-G-X3 in homologous recombination repair (HRR) is supported by our finding that FANCG and the RAD51-paralog XRCC3 are epistatic for sensitivity to DNA crosslinking compounds in DT40 chicken cells. Our findings further define the intricate interface between FANC and HRR proteins in maintaining chromosome stability.

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Acknowledgements

We thank all individuals who generously provided reagents, Yuxuan Xiao (NJJ), Keiko Namikoshi and Masayo Kimura (MT) for expert technical assistance, the Fanconi Anemia Research Fund for nurturing research collaborations and the North West Cancer Research Fund (UK) for financial support. This work was funded by grants NWCRF-CR624 and NWCRF-CR751 (NJJ). MT laboratory was supported in part by grants from the Ministry of Education, Culture, Sports, Science and Technology of Japan with financial support also provided by the Naito Foundation, and the Sagawa Foundation for Promotion of Cancer Research. SH and CGM acknowledge support from the Medical Research Council and the Daniel Ayling Fanconi Anemia Trust. The contribution by LHT was performed under the auspices of the US Department of Energy by Lawrence Livermore National Laboratory under Contract DE-AC52-07NA27344 and funded by the DOE Low-Dose Program and NCI/NIH grant CA112566. GMK acknowledges grant funding from NHLBI R01 HL063776.

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Author notes

  1. K Yamamoto

    Present address: 9Current address: Department of Hematology, Okayama Citizen's Hospital, Okayama City, Okayama 700-8557, Japan.,

  2. M Takata

    Present address: 10Current address: Radiation Biology Center, Kyoto University, Yoshida-konoe, Sakyo-ku, Kyoto 606-8501, Japan.,

Authors and Affiliations

  1. Molecular Oncology and Stem Cell Research Group, School of Biological Sciences, University of Liverpool, Liverpool, UK

    J B Wilson, A S Marriott & N J Jones

  2. Department of Immunology and Medical Genetics, Kawasaki Medical School, Kurashiki, Okayama, Japan

    K Yamamoto & M Takata

  3. Department of Biochemistry, University of Cambridge, Cambridge, UK

    S Hussain

  4. Department of Molecular Biophysics and Biochemistry, Yale University School of Medicine, New Haven, CT, USA

    P Sung

  5. Division of Biochemistry and Molecular Biology, Oregon Health Sciences University, Portland, OR, USA

    M E Hoatlin

  6. Department of Medical and Molecular Genetics, King's College London School of Medicine, Guy's Hospital, London, UK

    C G Mathew

  7. Biosciences and Biotechnology Division, L441, Lawrence Livermore National Laboratory, Livermore, CA, USA

    L H Thompson

  8. Department of Pediatrics, Division of Hematology-Oncology, Yale University School of Medicine, New Haven, CT, USA

    G M Kupfer

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Wilson, J., Yamamoto, K., Marriott, A. et al. FANCG promotes formation of a newly identified protein complex containing BRCA2, FANCD2 and XRCC3. Oncogene 27, 3641–3652 (2008). https://doi.org/10.1038/sj.onc.1211034

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