Direct interaction of FANCD2 with BRCA2 in DNA damage response pathways

Shobbir Hussain, James B. Wilson, Annette L. Medhurst, James Hejna, Emily Witt, Sahana Ananth, Adelina Davies, Jean Yves Masson, Robb Moses, Stephen C. West, Johan P. de Winter, Alan Ashworth, Nigel J. Jones, Christopher G. Mathew

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164 Citations (Scopus)

Abstract

Fanconi anaemia (FA) is a chromosomal instability disorder characterized by cellular sensitivity to DNA interstrand crosslinking agents and a high risk of cancer. Six of the eight proteins encoded by the known FA genes form a nuclear complex which is required for the monoubiquitination of the FANCD2 protein. FANCD2 complexes and colocalizes with BRCA1, but its presumptive role in DNA repair has not yet been clearly defined. We used yeast two-hybrid analysis to test for interaction between FANCD2 and 10 proteins involved in homologous recombination repair. FANCD2 did not interact with RAD51, the five RAD51 paralogs, RAD52, RAD54 or DMC1. However, it bound to a highly conserved C-terminal site in BRCA2 that also binds FANCG/XRCC9. FANCD2 and BRCA2 can be coimmunoprecipitated from cell extracts of both human and Chinese hamster wild-type cells, thus confirming that the interaction occurs in vivo. Formation of nuclear foci of FANCD2 was normal in the BRCA2 mutant CAPAN-1 cells, which indicates that the recruitment of FANCD2 to sites of DNA-repair is independent of wild-type BRCA2 function. FANCD2 colocalized with RAD51 in foci following treatment with mitomycin C or hydroxyurea, and colocalized very tightly with PCNA after treatment with hydroxyurea. These findings suggest that FANCD2 may have a role in the cellular response to stalled replication forks or in the repair of replication-associated double-strand breaks, irrespective of the type of primary DNA lesion.

Original languageEnglish (US)
Pages (from-to)1241-1248
Number of pages8
JournalHuman Molecular Genetics
Volume13
Issue number12
DOIs
StatePublished - Jun 15 2004

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Fanconi Anemia Complementation Group D2 Protein
Fanconi Anemia
Hydroxyurea
DNA Repair
DNA Damage
Chromosome Disorders
Recombinational DNA Repair
Chromosomal Instability
DNA
Proliferating Cell Nuclear Antigen
Mitomycin
Cricetulus
Cell Extracts
Yeasts
Genes
Neoplasms
Proteins

ASJC Scopus subject areas

  • Genetics

Cite this

Hussain, S., Wilson, J. B., Medhurst, A. L., Hejna, J., Witt, E., Ananth, S., ... Mathew, C. G. (2004). Direct interaction of FANCD2 with BRCA2 in DNA damage response pathways. Human Molecular Genetics, 13(12), 1241-1248. https://doi.org/10.1093/hmg/ddh135

Direct interaction of FANCD2 with BRCA2 in DNA damage response pathways. / Hussain, Shobbir; Wilson, James B.; Medhurst, Annette L.; Hejna, James; Witt, Emily; Ananth, Sahana; Davies, Adelina; Masson, Jean Yves; Moses, Robb; West, Stephen C.; de Winter, Johan P.; Ashworth, Alan; Jones, Nigel J.; Mathew, Christopher G.

In: Human Molecular Genetics, Vol. 13, No. 12, 15.06.2004, p. 1241-1248.

Research output: Contribution to journalArticle

Hussain, S, Wilson, JB, Medhurst, AL, Hejna, J, Witt, E, Ananth, S, Davies, A, Masson, JY, Moses, R, West, SC, de Winter, JP, Ashworth, A, Jones, NJ & Mathew, CG 2004, 'Direct interaction of FANCD2 with BRCA2 in DNA damage response pathways', Human Molecular Genetics, vol. 13, no. 12, pp. 1241-1248. https://doi.org/10.1093/hmg/ddh135
Hussain S, Wilson JB, Medhurst AL, Hejna J, Witt E, Ananth S et al. Direct interaction of FANCD2 with BRCA2 in DNA damage response pathways. Human Molecular Genetics. 2004 Jun 15;13(12):1241-1248. https://doi.org/10.1093/hmg/ddh135
Hussain, Shobbir ; Wilson, James B. ; Medhurst, Annette L. ; Hejna, James ; Witt, Emily ; Ananth, Sahana ; Davies, Adelina ; Masson, Jean Yves ; Moses, Robb ; West, Stephen C. ; de Winter, Johan P. ; Ashworth, Alan ; Jones, Nigel J. ; Mathew, Christopher G. / Direct interaction of FANCD2 with BRCA2 in DNA damage response pathways. In: Human Molecular Genetics. 2004 ; Vol. 13, No. 12. pp. 1241-1248.
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AU - Ananth, Sahana

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