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Mammalian XRCC2 promotes the repair of DNA double-strand breaks by homologous recombination

Nature volume 401, pages 397–399 (1999)Cite this article

Abstract

The repair of DNA double-strand breaks is essential for cells to maintain their genomic integrity. Two major mechanisms are responsible for repairing these breaks in mammalian cells, non-homologous end-joining (NHEJ) and homologous recombination (HR)1,2: the importance of the former in mammalian cells is well established3, whereas the role of the latter is just emerging. Homologous recombination is presumably promoted by an evolutionarily conserved group of genes termed the Rad52 epistasis group4,5,6,7,8,9,10,11. An essential component of the HR pathway is the strand-exchange protein, known as RecA in bacteria8 or Rad51 in yeast6. Several mammalian genes have been implicated in repair by homologous recombination on the basis of their sequence homology to yeast Rad51 (ref. 11): one of these is human XRCC2 (refs 12, 13). Here we show that XRCC2 is essential for the efficient repair of DNA double-strand breaks by homologous recombination between sister chromatids. We find that hamster cells deficient in XRCC2 show more than a 100-fold decrease in HR induced by double-strand breaks compared with the parental cell line. This defect is corrected to almost wild-type levels by transient transfection with a plasmid expressing XRCC2. The repair defect in XRCC2 mutant cells appears to be restricted to recombinational repair because NHEJ is normal. We conclude that XRCC2 is involved in the repair of DNA double-strand breaks by homologous recombination.

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Figure 1: Recombination reporter substrate SCneo.
Figure 2: XRCC2-deficient irs1 cells have severely reduced levels of DSB repair by HR.
Figure 3: XRCC2-deficient irs1 cells have normal levels of NHEJ.

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References

  1. Rouet,P., Smith,F. & Jasin,M. Introduction of double-strand breaks into the genome of mouse cells by expression of a rare-cutting endonuclease. Mol. Cell. Biol. 14, 8096–8106 (1994).

    Article  CAS  Google Scholar 

  2. Liang,F., Han,M., Romanienko,P. J. & Jasin,M. Homology-directed repair is a major double-strand break repair pathway in mammalian cells. Proc. Natl Acad. Sci. USA 95, 5172–5177 (1998).

    Article  ADS  CAS  Google Scholar 

  3. Jeggo,P. A. DNA breakage and repair. Adv. Gent. 38, 185–218 (1998).

    Article  CAS  Google Scholar 

  4. Bezzubova,O., Silbergleit,A., Yamaguchi-Iwai,Y., Takeda,S. & Buerstedde,J. M. Reduced X-ray resistance and homologous recombination frequencies in a RAD54-/- mutant of the chicken DT40 cell line. Cell 89, 185–193 (1997).

    Article  CAS  Google Scholar 

  5. Essers,J. et al. Disruption of mouse RAD54 reduces ionizing radiation resistance and homologous recombination. Cell 89, 195–204 (1997).

    Article  CAS  Google Scholar 

  6. Nickoloff,J. A. & Hoekstra,M. F. in DNA Damage and Repair Vol. I (eds Nickoloff, J. A. & Hoekstra, M. F.) 335–362 (Humana, Totowa, 1998).

    Google Scholar 

  7. Rijkers,T. et al. Targeted inactivation of mouse RAD52 reduces homologous recombination but not resistance to ionizing radiation. Mol. Cell. Biol. 18, 6423–6429 (1998).

    Article  CAS  Google Scholar 

  8. Smith,G. R. in DNA Damage and Repair Vol. I (eds Nickoloff, J. A. & Hoekstra, M. F.) 135–162 (Humana, Totowa, 1998).

    Book  Google Scholar 

  9. Sonoda,E. et al. Rad51-deficient vertebrate cells accumulate chromosomal breaks prior to cell death. EMBO J. 17, 598–608 (1998).

    Article  CAS  Google Scholar 

  10. Yamaguchi-Iwai,Y. et al. Homologous recombination, but not DNA repair, is reduced in vertebrate cells deficient in RAD52. Mol. Cell. Biol. 18, 6430–6435 (1998).

    Article  CAS  Google Scholar 

  11. Thacker,J. A surfeit of RAD51-like genes? Trends Genet. 15, 166–168 (1999).

    Article  CAS  Google Scholar 

  12. Cartwright,R., Tambini,C. E., Simpson,P. J. & Thacker,J. The XRCC2 DNA repair gene from human and mouse encodes a novel member of the recA/RAD51 family. Nucleic Acids Res. 26, 3084–3089 (1998).

    Article  CAS  Google Scholar 

  13. Liu,N. et al. XRCC2 and XRCC3, new human Rad51-family members, promote chromosome stability and protect against DNA cross-links and other damages. Molecular Cell 1, 783–793 (1998).

    Article  CAS  Google Scholar 

  14. Jones,N. J., Cox,R. & Thacker,J. Isolation and cross-sensitivity of X-ray-sensitive mutants of V79-4 hamster cells. Mutat. Res. 183, 279–286 (1987).

    CAS  PubMed  Google Scholar 

  15. Liang,F., Romanienko,P. J., Weaver,D. T., Jeggo,P. A. & Jasin,M. Chromosomal double-strand break repair in Ku80 deficient cells. Proc. Natl Acad. Sci. USA 93, 8929–8933 (1996).

    Article  ADS  CAS  Google Scholar 

  16. Smih,F., Rouet,P., Romanienko,P. J. & Jasin,M. Double-strand breaks at the target locus stimulate gene targeting in embryonic stem cells. Nucleic Acids Res. 23, 5012–5019 (1995).

    Article  CAS  Google Scholar 

  17. Jasin,M. Double-strand break repair and homologous recombination in mammalian cells. in DNA Damage and Repair Vol. III (eds Nickoloff, J. A. & Hoekstra, M. F.) (Humana, Totowa, in the press).

  18. Cheong,N., Wang,X., Wang,Y. & Iliakis,G. Loss of S-phase-dependent radioresistance in irs-1 cells exposed to X-rays. Mutat. Res. 314, 77–85 (1994).

    Article  CAS  Google Scholar 

  19. Tsuzuki,T. et al. Targeted disruption of the Rad51 gene leads to lethality in embryonic mice. Proc. Natl Acad. Sci. USA 93, 6236–6340 (1996).

    Article  ADS  CAS  Google Scholar 

  20. Lim, D.-S. & Hasty,P. A mutation in mouse rad51 results in an early embryonic lethal that is suppressed by a mutation in p53. Mol. Cell. Biol. 16, 7133–7143 (1996).

    Article  Google Scholar 

  21. Baumann,P., Benson,F. E. & West,S. C. Human Rad51 protein promotes ATP dependent homologous pairing and strand transfer reactions in vitro. Cell 87, 757–766 (1996).

    Article  CAS  Google Scholar 

  22. Dosanjh,M. K. et al. Isolation and characterization of RAD51C a new human member of the RAD51 family of related genes. Nucleic Acids Res. 26, 1179–1184 (1998).

    Article  CAS  Google Scholar 

  23. Hays,S. L., Firmenich,A. A. & Berg,P. Complex formation in yeast double-strand break repair: Participation of Rad51, Rad52, Rad55, and Rad57 proteins. Proc. Natl Acad. Sci. USA 92, 6925–6929 (1995).

    Article  ADS  CAS  Google Scholar 

  24. Johnson,R. D. & Symington,L. S. Functional differences and interactions among the putative RecA homologs Rad51, Rad55, and Rad57. Mol. Cell. Biol. 15, 4843–4850 (1995).

    Article  CAS  Google Scholar 

  25. Sung,P. Yeast Rad55 and Rad57 proteins form a heterodimer that functions with replication protein A to promote DNA strand exchange by Rad51 recombinase. Genes Dev. 11, 1111–1121 (1997).

    Article  CAS  Google Scholar 

  26. Vogelstein,B. & Kinzler,K. W. (eds) The Genetic Basis of Human Cancer (MacGraw-Hill, New York, 1998).

    Google Scholar 

  27. Kinzler,K. W. & Vogelstein,B. Cancer-susceptibility genes. Gatekeepers and caretakers. Nature 386, 761, 763 (1997).

    Article  Google Scholar 

  28. Rouet,P., Smih,F. & Jasin,M. Expression of a site-specific endonuclease stimulates homologous recombination in mammalian cells. Proc. Natl Acad. Sci. USA 91, 6064–6068 (1994).

    Article  ADS  CAS  Google Scholar 

  29. Donoho,G., Jasin,M. & Berg,P. Analysis of gene targeting and intrachromosomal homologous recombination stimulated by genomic double-strand breaks in mouse embryonic stem cells. Mol. Cell. Biol. 18, 4070–4078 (1998).

    Article  CAS  Google Scholar 

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Acknowledgements

We thank L. Thompson and N. Jones for their assistance. This work was funded by an NRSA fellowship to R.D.J. a DOE grant (N.L.) and an NIH grant to M.J.

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Authors and Affiliations

  1. Cell Biology Program, Memorial Sloan-Kettering Cancer Center, and Cornell University Graduate School of Medical Sciences, 1275 York Avenue, New York, 10021, New York, USA

    Roger D. Johnson & Maria Jasin

  2. Biology and Biotechnology Research Program, Lawrence Livermore National Laboratory, Livermore, 94551, California, USA

    Nan Liu

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  1. Roger D. Johnson
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Correspondence to Maria Jasin.

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Johnson, R., Liu, N. & Jasin, M. Mammalian XRCC2 promotes the repair of DNA double-strand breaks by homologous recombination. Nature 401, 397–399 (1999). https://doi.org/10.1038/43932

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