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Human Molecular Genetics Advance Access originally published online on August 5, 2003
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Human Molecular Genetics, 2003, Vol. 12, No. 19 2503-2510
DOI: 10.1093/hmg/ddg266
© 2003 Oxford University Press

Direct interaction of the Fanconi anaemia protein FANCG with BRCA2/FANCD1

Shobbir Hussain1, Emily Witt2, Pia A.J. Huber1, Annette L. Medhurst1, Alan Ashworth2 and Christopher G. Mathew1,*

1Division of Genetics and Development, Guy's, King's and St Thomas' School of Medicine, King's College London, 8th Floor, Guy's Tower, Guy's Hospital, London SE1 9RT, UK and 2Cancer Research UK Gene Function and Regulation Group, The Breakthrough Breast Cancer Research Centre, The Institute of Cancer Research, Fulham Road, London SW3 6JB, UK

Received May 9, 2003; Revised July 15, 2003; Accepted July 30, 2003

Fanconi anaemia (FA) is an autosomal recessive genetic disorder characterized by progressive bone marrow failure, multiple congenital abnormalities, and an increased risk of cancer. FA cells are characterized by chromosomal instability and hypersensitivity to DNA interstrand crosslinking agents. At least eight complementation groups exist (FA-A to G), and the genes for all of these except FA-B have been cloned. Functional linkage between the FA pathway and genes involved in susceptibility to breast cancer has been demonstrated by the interaction of the FANCA and FANCD2 proteins with BRCA1, and the discovery that the FANCD1 gene is identical to BRCA2. Here we have used the yeast two-hybrid system to test for direct interaction between BRCA2 or its effector RAD51 and the FANCA, FANCC and FANCG proteins. We found that FANCG was capable of binding to two separate sites in the BRCA2 protein, located either side of the BRC repeats. Furthermore, FANCG could be co-immunoprecipitated with BRCA2 from human cells, and FANCG co-localized in nuclear foci with both BRCA2 and RAD51 following DNA damage with mitomycin C. These results demonstrate that BRCA2 is directly connected to a pathway that is deficient in interstrand crosslink repair, and that at least one other FA protein is closely associated with the homologous recombination DNA repair machinery.

* To whom correspondence should be addressed. Tel: +44 2079554653; Fax: +44 2079554644; Email: christopher.mathew{at}kcl.ac.uk


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