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The BRIP1 helicase functions independently of BRCA1 in the Fanconi anemia pathway for DNA crosslink repair

Abstract

BRIP1 (also called BACH1) is a DEAH helicase that interacts with the BRCT domain of BRCA1 (refs. 16) and has an important role in BRCA1-dependent DNA repair and checkpoint functions1,2,6,7. We cloned the chicken ortholog of BRIP1 and established a homozygous knockout in the avian B-cell line DT40. The phenotype of these brip1 mutant cells in response to DNA damage differs from that of brca1 mutant cells and more closely resembles that of fancc mutant cells, with a profound sensitivity to the DNA-crosslinking agent cisplatin and acute cell-cycle arrest in late S-G2 phase. These defects are corrected by expression of human BRIP1 lacking the BRCT-interaction domain. Moreover, in human cells exposed to mitomycin C, short interfering RNA–mediated knock-down of BRIP1 leads to a substantial increase in chromosome aberrations, a characteristic phenotype of cells derived from individuals with Fanconi anemia. Because brip1 mutant cells are proficient for ubiquitination of FANCD2 protein, our data indicate that BRIP1 has a function in the Fanconi anemia pathway that is independent of BRCA1 and downstream of FANCD2 activation.

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Figure 1: Generation of a homozygous brip1 knockout in the DT40 cell line.
Figure 2: Sensitivity of brip1 and brca1 mutant cells to DNA-damaging agents.
Figure 3: brip1 mutant cells are proficient for HDR of an I-SceI–induced DNA double-strand break.
Figure 4: brip1 cells, but not brca1 cells, arrest with a DNA content near 4N after exposure to cisplatin.
Figure 5: brip1 cells have increased chromosome instability.

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References

  1. Cantor, S.B. et al. BACH1, a novel helicase-like protein, interacts directly with BRCA1 and contributes to its DNA repair function. Cell 105, 149–160 (2001).

    Article  CAS  Google Scholar 

  2. Yu, X., Chini, C.C., He, M., Mer, G. & Chen, J. The BRCT domain is a phospho-protein binding domain. Science 302, 639–642 (2003).

    Article  CAS  Google Scholar 

  3. Cantor, S. et al. The BRCA1-associated protein BACH1 is a DNA helicase targeted by clinically relevant inactivating mutations. Proc. Natl. Acad. Sci. USA 101, 2357–2362 (2004).

    Article  CAS  Google Scholar 

  4. Williams, R.S., Lee, M.S., Hau, D.D. & Glover, J.N. Structural basis of phosphopeptide recognition by the BRCT domain of BRCA1. Nat. Struct. Mol. Biol. 11, 519–525 (2004).

    Article  CAS  Google Scholar 

  5. Clapperton, J.A. et al. Structure and mechanism of BRCA1 BRCT domain recognition of phosphorylated BACH1 with implications for cancer. Nat. Struct. Mol. Biol. 11, 512–518 (2004).

    Article  CAS  Google Scholar 

  6. Botuyan, M.V. et al. Structural basis of BACH1 phosphopeptide recognition by BRCA1 tandem BRCT domains. Structure (Camb.) 12, 1137–1146 (2004).

    Article  CAS  Google Scholar 

  7. Yu, X. & Chen, J. DNA damage-induced cell cycle checkpoint control requires CtIP, a phosphorylation-dependent binding partner of BRCA1 C-terminal domains. Mol. Cell. Biol. 24, 9478–9486 (2004).

    Article  CAS  Google Scholar 

  8. Manke, I.A., Lowery, D.M., Nguyen, A. & Yaffe, M.B. BRCT repeats as phosphopeptide-binding modules involved in protein targeting. Science 302, 636–639 (2003).

    Article  CAS  Google Scholar 

  9. Shiozaki, E.N., Gu, L., Yan, N. & Shi, Y. Structure of the BRCT repeats of BRCA1 bound to a BACH1 phosphopeptide: implications for signaling. Mol. Cell 14, 405–412 (2004).

    Article  CAS  Google Scholar 

  10. Moynahan, M.E., Chiu, J.W., Koller, B.H. & Jasin, M. Brca1 controls homology-directed DNA repair. Mol. Cell 4, 511–518 (1999).

    Article  CAS  Google Scholar 

  11. Pierce, A.J., Johnson, R.D., Thompson, L.H. & Jasin, M. XRCC3 promotes homology-directed repair of DNA damage in mammalian cells. Genes Dev. 13, 2633–2638 (1999).

    Article  CAS  Google Scholar 

  12. Yarden, R.I., Pardo-Reoyo, S., Sgagias, M., Cowan, K.H. & Brody, L.C. BRCA1 regulates the G2/M checkpoint by activating Chk1 kinase upon DNA damage. Nat. Genet. 30, 285–289 (2002).

    Article  Google Scholar 

  13. Xu, B., Kim, S. & Kastan, M.B. Involvement of Brca1 in S-phase and G(2)-phase checkpoints after ionizing irradiation. Mol. Cell. Biol. 21, 3445–3450 (2001).

    Article  CAS  Google Scholar 

  14. Poll, E.H., Arwert, F., Joenje, H. & Eriksson, A.W. Cytogenetic toxicity of antitumor platinum compounds in Fanconi's anemia. Hum. Genet. 61, 228–230 (1982).

    Article  CAS  Google Scholar 

  15. Simpson, L.J. & Sale, J.E. Rev1 is essential for DNA damage tolerance and non-templated immunoglobulin gene mutation in a vertebrate cell line. EMBO J. 22, 1654–1664 (2003).

    Article  CAS  Google Scholar 

  16. Yamashita, Y.M. et al. RAD18 and RAD54 cooperatively contribute to maintenance of genomic stability in vertebrate cells. EMBO J. 21, 5558–5566 (2002).

    Article  CAS  Google Scholar 

  17. Auerbach, A.D. & Wolman, S.R. Susceptibility of Fanconi's anaemia fibroblasts to chromosome damage by carcinogens. Nature 261, 494–496 (1976).

    Article  CAS  Google Scholar 

  18. Ishida, R. & Buchwald, M. Susceptibility of Fanconi's anemia lymphoblasts to DNA-cross-linking and alkylating agents. Cancer Res. 42, 4000–4006 (1982).

    CAS  PubMed  Google Scholar 

  19. Sasaki, M.S. & Tonomura, A. A high susceptibility of Fanconi's anemia to chromosome breakage by DNA cross-linking agents. Cancer Res. 33, 1829–1836 (1973).

    CAS  PubMed  Google Scholar 

  20. Joenje, H. & Patel, K.J. The emerging genetic and molecular basis of Fanconi anaemia. Nat. Rev. Genet. 2, 446–457 (2001).

    Article  CAS  Google Scholar 

  21. Niedzwiedz, W. et al. The Fanconi anaemia gene FANCC promotes homologous recombination and error-prone DNA repair. Mol. Cell 15, 607–620 (2004).

    Article  CAS  Google Scholar 

  22. Joenje, H., Arwert, F., Eriksson, A.W., de Koning, H. & Oostra, A.B. Oxygen-dependence of chromosomal aberrations in Fanconi's anaemia. Nature 290, 142–143 (1981).

    Article  CAS  Google Scholar 

  23. Garcia-Higuera, I. et al. Interaction of the Fanconi anemia proteins and BRCA1 in a common pathway. Mol. Cell 7, 249–262 (2001).

    Article  CAS  Google Scholar 

  24. Levitus, M. et al. Heterogeneity in Fanconi anemia: evidence for 2 new genetic subtypes. Blood 103, 2498–2503 (2004).

    Article  CAS  Google Scholar 

  25. Levitus, M. et al. The DNA helicase BRIP1 is defective in Fanconi anemia complementation group J. Nat. Genet., advance online publication 21 August 2005 (10.1038/ng1625).

  26. Boardman, P.E. et al. A comprehensive collection of chicken cDNAs. Curr. Biol. 12, 1965–1969 (2002).

    Article  Google Scholar 

  27. Arakawa, H., Lodygin, D. & Buerstedde, J.M. Mutant loxP vectors for selectable marker recycle and conditional knock-outs. BMC Biotechnol. 1, 7 (2001).

    Article  CAS  Google Scholar 

  28. Vandenberg, C.J. et al. BRCA1-independent ubiquitination of FANCD2. Mol. Cell 12, 247–254 (2003).

    Article  CAS  Google Scholar 

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Acknowledgements

We thank D. Livingston for human BRIP1 cDNA; J. Sale for DT40 cells; J. Di Noia for DT40 cells with an integrated DR-GFP; K.J. Patel for antibody to FANCD2, fancc mutant cells and FANCC disruption construct; A. Oostra for assistance with chromosome breakage assays in HeLa cells; H. Joenje and J. de Winter for advice and for communicating data before publication; and our colleagues at the Laboratory of Molecular Biology for advice and technical assistance. C.J.V. was funded in part by a grant from the Association for International Cancer Research.

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Correspondence to Kevin Hiom.

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Supplementary information

Supplementary Fig. 1

Clustal alignment of human (Hs) and chicken (Gg) BRIP1 protein sequences. (PDF 118 kb)

Supplementary Table 1

Oligonucleotide primers used for PCR as described in Methods. (PDF 26 kb)

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Bridge, W., Vandenberg, C., Franklin, R. et al. The BRIP1 helicase functions independently of BRCA1 in the Fanconi anemia pathway for DNA crosslink repair. Nat Genet 37, 953–957 (2005). https://doi.org/10.1038/ng1627

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