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Molecular and Cellular Biology, August 2003, p. 5421-5430, Vol. 23, No. 15
0270-7306/03/$08.00+0     DOI: 10.1128/MCB.23.15.5421-5430.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Fanconi Anemia FANCG Protein in Mitigating Radiation- and Enzyme-Induced DNA Double-Strand Breaks by Homologous Recombination in Vertebrate Cells

Kazuhiko Yamamoto,1,2 Masamichi Ishiai,1 Nobuko Matsushita,1 Hiroshi Arakawa,3 Jane E. Lamerdin,4 Jean-Marie Buerstedde,3 Mitsune Tanimoto,2 Mine Harada,5 Larry H. Thompson,4 and Minoru Takata1*

Department of Immunology and Molecular Genetics, Kawasaki Medical School, Kurashiki, Okayama 701-0192,1 Department of Internal Medicine, Okayama University Medical School, Okayama 700-8558,2 Department of Internal Medicine, Kyushu University School of Medicine, Fukuoka 812-8582, Japan,5 Institute for Molecular Radiobiology, GSF, D-85764 Neuherberg Munich, Germany,3 Biology and Biotechnology Research Program, Lawrence Livermore National Laboratory, Livermore, California 94551-08084

Received 2 December 2002/ Returned for modification 26 January 2003/ Accepted 24 April 2003

The rare hereditary disorder Fanconi anemia (FA) is characterized by progressive bone marrow failure, congenital skeletal abnormality, elevated susceptibility to cancer, and cellular hypersensitivity to DNA cross-linking chemicals and sometimes other DNA-damaging agents. Molecular cloning identified six causative genes (FANCA, -C, -D2, -E, -F, and -G) encoding a multiprotein complex whose precise biochemical function remains elusive. Recent studies implicate this complex in DNA damage responses that are linked to the breast cancer susceptibility proteins BRCA1 and BRCA2. Mutations in BRCA2, which participates in homologous recombination (HR), are the underlying cause in some FA patients. To elucidate the roles of FA genes in HR, we disrupted the FANCG/XRCC9 locus in the chicken B-cell line DT40. FANCG-deficient DT40 cells resemble mammalian fancg mutants in that they are sensitive to killing by cisplatin and mitomycin C (MMC) and exhibit increased MMC and radiation-induced chromosome breakage. We find that the repair of I-SceI-induced chromosomal double-strand breaks (DSBs) by HR is decreased ~9-fold in fancg cells compared with the parental and FANCG-complemented cells. In addition, the efficiency of gene targeting is mildly decreased in FANCG-deficient cells, but depends on the specific locus. We conclude that FANCG is required for efficient HR-mediated repair of at least some types of DSBs.

* Corresponding author. Mailing address: Department of Immunology and Molecular Genetics, Kawasaki Medical School, Kurashiki, Okayama 701-0192, Japan. Phone: 81-86-462-1111. Fax: 81-86-464-1187. E-mail: mtakata{at}med.kawasaki-m.ac.jp.

Molecular and Cellular Biology, August 2003, p. 5421-5430, Vol. 23, No. 15
0022-538X/03/$08.00+0     DOI: 10.1128/MCB.23.15.5421-5430.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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