Association of BRCA1 with the hRad50-hMre11-p95 Complex and the DNA Damage Response
10.1126/science.285.5428.747
Crossref journal-article
American Association for the Advancement of Science (AAAS)
Science (221)
Abstract

BRCA1 encodes a tumor suppressor that is mutated in familial breast and ovarian cancers. Here, it is shown that BRCA1 interacts in vitro and in vivo with hRad50, which forms a complex with hMre11 and p95/nibrin. Upon irradiation, BRCA1 was detected in discrete foci in the nucleus, which colocalize with hRad50. Formation of irradiation-induced foci positive for BRCA1, hRad50, hMre11, or p95 was dramatically reduced in HCC/1937 breast cancer cells carrying a homozygous mutation in BRCA1 but was restored by transfection of wild-type BRCA1 . Ectopic expression of wild-type, but not mutated, BRCA1 in these cells rendered them less sensitive to the DNA damage agent, methyl methanesulfonate. These data suggest that BRCA1 is important for the cellular responses to DNA damage that are mediated by the hRad50-hMre11-p95 complex.

Bibliography

Zhong, Q., Chen, C.-F., Li, S., Chen, Y., Wang, C.-C., Xiao, J., Chen, P.-L., Sharp, Z. D., & Lee, W.-H. (1999). Association of BRCA1 with the hRad50-hMre11-p95 Complex and the DNA Damage Response. Science, 285(5428), 747–750.

Authors 9
  1. Qing Zhong (first)
  2. Chi-Fen Chen (additional)
  3. Shang Li (additional)
  4. Yumay Chen (additional)
  5. Chuan-Cheng Wang (additional)
  6. Jun Xiao (additional)
  7. Phang-Lang Chen (additional)
  8. Z. Dave Sharp (additional)
  9. Wen-Hwa Lee (additional)
References 42 Referenced 469
  1. 10.1126/science.2270482
  2. Miki Y. et al. 266 66 (1994).
  3. 10.1074/jbc.273.39.25388
  4. Wong A. K., et al., Oncogene 17, 2279 (1998); (10.1038/sj.onc.1202150) / Oncogene by Wong A. K. (1998)
  5. Li S., et al., J. Biol. Chem. 274, 11334 (1999). (10.1074/jbc.274.16.11334) / J. Biol. Chem. by Li S. (1999)
  6. 10.1126/science.281.5379.1009
  7. 10.1016/S0092-8674(00)80503-6
  8. Scully R. et al. 88 265 (1997). (10.1016/S0092-8674(00)81847-4)
  9. 10.1016/S1097-2765(00)80466-9
  10. 10.1126/science.270.5237.789
  11. Chen Y., et al., Cancer Res. 56, 3168 (1996). / Cancer Res. by Chen Y. (1996)
  12. Polyclonal antibodies were obtained by using the following bacterially expressed and purified GST fusion proteins as antigens: GST-hRad50-15A5 containing amino acids 211 through 575 of hRad50; GST-MM containing amino acids 82 through 582 of hMre11; GST-NBS1 containing amino acids 12 through 754 of p95; GST-hRad51 containing full-length hRad51; and GST alone (for anti-GST mAb 8G11) to generate polyclonal or monoclonal antibodies (25). BRCA1 mAb's 6B4 and 17F8 were described in (18). BRCA1 mAb Ab-1 and rabbit α-hRad51 antibodies were from Oncogene Research Product (Cambridge MA). Affinity-purified rabbit α-BRCA1 antibody (C-20) and α-HA antibody (Y-11) were from Santa Cruz Biotechnology (Santa Cruz CA).
  13. Haber J. E., Cell 95, 583 (1998). (10.1016/S0092-8674(00)81626-8) / Cell by Haber J. E. (1998)
  14. Raymond W. E., Kleckner N., Nucleic Acids Res. 21, 3851 (1993). (10.1093/nar/21.16.3851) / Nucleic Acids Res. by Raymond W. E. (1993)
  15. Paull T. T., Gellert M., Mol. Cell 1, 969 (1998); (10.1016/S1097-2765(00)80097-0) / Mol. Cell by Paull T. T. (1998)
  16. Trujillo K. M., Yuan S. S., Lee E. Y., Sung P., J. Biol. Chem. 273, 21447 (1998); (10.1074/jbc.273.34.21447) / J. Biol. Chem. by Trujillo K. M. (1998)
  17. Furuse M., et al., EMBO J. 17, 6412 (1998); (10.1093/emboj/17.21.6412) / EMBO J. by Furuse M. (1998)
  18. 10.1016/S0092-8674(00)81640-2
  19. Moreau S., Ferguson J. R., Symington L. S., Mol. Cell. Biol. 19, 556 (1999). (10.1128/MCB.19.1.556) / Mol. Cell. Biol. by Moreau S. (1999)
  20. Shiloh Y., Annu. Rev. Genet. 31, 635 (1997); (10.1146/annurev.genet.31.1.635) / Annu. Rev. Genet. by Shiloh Y. (1997)
  21. Weemaes C. M., Smeets D. F., van der Burgt C. J., Int. J. Radiat. Biol. 66, S185 (1994); (10.1080/09553009414552021) / Int. J. Radiat. Biol. by Weemaes C. M. (1994)
  22. van der Burgt I., Chrzanowska K. H., Smeets D., Weemaes C., J. Med. Genet. 33, 153 (1996); (10.1136/jmg.33.2.153) / J. Med. Genet. by van der Burgt I. (1996)
  23. Varon R., et al., Cell 93, 467 (1998). (10.1016/S0092-8674(00)81174-5) / Cell by Varon R. (1998)
  24. Carney J. P., et al., Cell 93, 477 (1998). (10.1016/S0092-8674(00)81175-7) / Cell by Carney J. P. (1998)
  25. 10.1126/science.280.5363.590
  26. Dolganov G. M., et al., Mol. Cell. Biol. 16, 4832 (1996). (10.1128/MCB.16.9.4832) / Mol. Cell. Biol. by Dolganov G. M. (1996)
  27. Maser R. S. Monsen K. J. Nelms B. E. Petrini J. H. J. 17 6087 (1997). (10.1128/MCB.17.10.6087)
  28. Q. Zhong and W.-H. Lee unpublished results.
  29. The procedure for indirect immunofluorescence staining was adopted and slightly modified from that in (25). In a typical staining experiment 500 to 1000 cells were counted. For the foci-containing cells only nuclei containing >10 foci were counted as a foci-containing cell. Each experiment was repeated at least three times and the results were reproducible with the antibodies indicated here. The results did not vary according to the conditions of fixation. The foci formation studies were performed following different time courses after irradiation (at 1 3 6 8 and 12 hours). Because all of the studied foci show relatively high quality and quantity at 8 hours after irradiation the results at this time point are mostly used as representatives here.
  30. H. K. Chew A. A. Farmer W.-H. Lee in Breast Cancer A. Bowcock Ed. (Humana Totowa NJ 1998) pp. 225–246. (10.1007/978-1-59259-456-6_10)
  31. Tomlinson G. E., et al., Cancer Res. 58, 3237 (1998). / Cancer Res. by Tomlinson G. E. (1998)
  32. Wu L. J. C., et al., Nature Genet. 14, 430 (1996). (10.1038/ng1296-430) / Nature Genet. by Wu L. J. C. (1996)
  33. Chapman M. S., Verma I. M., Nature 382, 678 (1996). (10.1038/382678a0) / Nature by Chapman M. S. (1996)
  34. Sharan S. K. et al. 386 804 (1997); (10.1038/386804a0)
  35. 10.1073/pnas.95.9.5287
  36. 10.1016/S1097-2765(00)80276-2
  37. 10.1038/25292
  38. Zhang H., Tombline G., Weber B., Cell 92, 433 (1998). (10.1016/S0092-8674(00)80936-8) / Cell by Zhang H. (1998)
  39. Zhong Q., et al., Cancer Res. 57, 4225 (1997). / Cancer Res. by Zhong Q. (1997)
  40. Synchronization of T24 human bladder carcinoma cells was performed by density arrest then released at time zero by replating at a density of 2 × 10 6 cells per 10-cm plate. At various time points thereafter (12 hours for G 1 18 hours for G 1 /S 24 hours for S 33 hours for G 2 /M and 1 hour for arrest in G 0 /G 1 ) cells were harvested. To obtain cells in M phase we added nocodazole (0.4 μg/ml) to the culture medium for 10 hours before harvest.
  41. Constructs based on plasmid pcDNA3.1 (Invitrogen San Diego CA) were used for lipofectin-mediated transfection of HCC1937 cells with BRCA1 cDNA. Cells were harvested after 48 hours for immunoprecipitation and protein immunoblot analysis. Parallel cultures were treated with 0.1% MMS for 50 min and surviving cells were counted after 8 days. The experiments were repeated at least three times and the results were consistent.
  42. We thank J. Petrini for rabbit antisera to hRad50 P. Garza and D. Jones for antibody production and N. Ting and T. Boyer for critical reading. Supported by grants from NIH (CA 58183 and CA 30195) the McDermott endowment (W.H.L.) and the Susan G. Komen Foundation for Breast Cancer Research (P.L.C.).
Dates
Type When
Created 23 years, 1 month ago (July 27, 2002, 1:42 a.m.)
Deposited 1 year, 7 months ago (Jan. 12, 2024, 11:13 p.m.)
Indexed 4 weeks, 1 day ago (Aug. 6, 2025, 8:52 a.m.)
Issued 26 years, 1 month ago (July 30, 1999)
Published 26 years, 1 month ago (July 30, 1999)
Published Print 26 years, 1 month ago (July 30, 1999)
Funders 0

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@article{Zhong_1999, title={Association of BRCA1 with the hRad50-hMre11-p95 Complex and the DNA Damage Response}, volume={285}, ISSN={1095-9203}, url={http://dx.doi.org/10.1126/science.285.5428.747}, DOI={10.1126/science.285.5428.747}, number={5428}, journal={Science}, publisher={American Association for the Advancement of Science (AAAS)}, author={Zhong, Qing and Chen, Chi-Fen and Li, Shang and Chen, Yumay and Wang, Chuan-Cheng and Xiao, Jun and Chen, Phang-Lang and Sharp, Z. Dave and Lee, Wen-Hwa}, year={1999}, month=jul, pages={747–750} }