Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Article
  • Published:

MDC1 interacts with Rad51 and facilitates homologous recombination

Nature Structural & Molecular Biology volume 12, pages 902–909 (2005)Cite this article

Abstract

Mediator of DNA damage checkpoint protein-1 (MDC1) is a recently identified nuclear protein that participates in DNA-damage sensing and signaling. Here we report that knockdown of MDC1 by RNA interference results in cellular hypersensitivity to the DNA cross-linking agent mitomycin C and ionizing radiation and in impaired homology-mediated repair of double-strand breaks in DNA. MDC1 forms a complex with Rad51 through a direct interaction with the forkhead-associated domain of MDC1, not the BRCA1 C-terminal domain. Depletion of MDC1 results in impaired formation of Rad51 ionizing radiation–induced foci, reduced amounts of nuclear and chromatin-bound Rad51, and a corresponding increase in Rad51 protein degradation. Together, our findings suggest that MDC1 functions in Rad51-mediated homologous recombination by retaining Rad51 in chromatin.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on SpringerLink
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1: MDC1 is involved in DNA damage response to MMC and ionizing radiation (IR).
Figure 2: Impaired I-SceI–induced homologous recombination in MDC1-deficient cells.
Figure 3: Association of MDC1 and Rad51.
Figure 4: Rad51 IRIF are deficient in MDC1-knockdown cells at early time points after exposure to ionizing radiation (IR).
Figure 5: Reduction of nuclear and chromatin-bound Rad51 protein after MDC1 siRNA transfection.
Figure 6: Decreased Rad51 protein stability in cells with MDC1 knockdown.

Similar content being viewed by others

Accession codes

Accessions

BINDPlus

References

  1. van Gent, D.C., Hoeijmakers, J.H. & Kanaar, R. Chromosomal stability and the DNA double-stranded break connection. Nat. Rev. Genet. 2, 196–206 (2001).

    Article  CAS  Google Scholar 

  2. Thompson, L.H. & Schild, D. Homologous recombinational repair of DNA ensures mammalian chromosome stability. Mutat. Res. 477, 131–153 (2001).

    Article  CAS  Google Scholar 

  3. Xu, X. & Stern, D.F. NFBD1/MDC1 regulates ionizing radiation-induced focus formation by DNA checkpoint signaling and repair factors. FASEB J. 17, 1842–1848 (2003).

    Article  CAS  Google Scholar 

  4. Xu, X. & Stern, D.F. NFBD1/KIAA0170 is a chromatin-associated protein involved in DNA damage signaling pathways. J. Biol. Chem. 278, 8795–8803 (2003).

    Article  CAS  Google Scholar 

  5. Mochan, T.A., Venere, M., DiTullio, R.A., Jr. & Halazonetis, T.D. 53BP1 and NFBD1/MDC1-Nbs1 function in parallel interacting pathways activating ataxia-telangiectasia mutated (ATM) in response to DNA damage. Cancer Res. 63, 8586–8591 (2003).

    CAS  PubMed  Google Scholar 

  6. Stewart, G.S., Wang, B., Bignell, C.R., Taylor, A.M. & Elledge, S.J. MDC1 is a mediator of the mammalian DNA damage checkpoint. Nature 421, 961–966 (2003).

    Article  CAS  Google Scholar 

  7. Stucki, M. & Jackson, S.P. MDC1/NFBD1: a key regulator of the DNA damage response in higher eukaryotes. DNA Repair (Amst.) 3, 953–957 (2004).

    Article  CAS  Google Scholar 

  8. Goldberg, M. et al. MDC1 is required for the intra-S-phase DNA damage checkpoint. Nature 421, 952–956 (2003).

    Article  CAS  Google Scholar 

  9. Lou, Z. et al. MDC1 regulates DNA-PK autophosphorylation in response to DNA damage. J. Biol. Chem. 279, 46359–46362 (2004).

    Article  CAS  Google Scholar 

  10. Lou, Z., Chini, C.C., Minter-Dykhouse, K. & Chen, J. Mediator of DNA damage checkpoint protein 1 regulates BRCA1 localization and phosphorylation in DNA damage checkpoint control. J. Biol. Chem. 278, 13599–13602 (2003).

    Article  CAS  Google Scholar 

  11. Lou, Z., Minter-Dykhouse, K., Wu, X. & Chen, J. MDC1 is coupled to activated CHK2 in mammalian DNA damage response pathways. Nature 421, 957–961 (2003).

    Article  CAS  Google Scholar 

  12. Zhang, J. et al. Chk2 phosphorylation of BRCA1 regulates DNA double-strand break repair. Mol. Cell. Biol. 24, 708–718 (2004).

    Article  CAS  Google Scholar 

  13. Callebaut, I. & Mornon, J.P. From BRCA1 to RAP1: a widespread BRCT module closely associated with DNA repair. FEBS Lett. 400, 25–30 (1997).

    Article  CAS  Google Scholar 

  14. Hofmann, K. & Bucher, P. The FHA domain: a putative nuclear signalling domain found in protein kinases and transcription factors. Trends Biochem. Sci. 20, 347–349 (1995).

    Article  CAS  Google Scholar 

  15. Tsai, M.D. FHA: a signal transduction domain with diverse specificity and function. Structure (Camb). 10, 887–888 (2002).

    Article  CAS  Google Scholar 

  16. 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 

  17. Durocher, D. et al. The molecular basis of FHA domain: phosphopeptide binding specificity and implications for phospho-dependent signaling mechanisms. Mol. Cell 6, 1169–1182 (2000).

    Article  CAS  Google Scholar 

  18. Dimitrova, D.S. & Gilbert, D.M. Stability and nuclear distribution of mammalian replication protein A heterotrimeric complex. Exp. Cell Res. 254, 321–327 (2000).

    Article  CAS  Google Scholar 

  19. Vassin, V.M., Wold, M.S. & Borowiec, J.A. Replication protein A (RPA) phosphorylation prevents RPA association with replication centers. Mol. Cell. Biol. 24, 1930–1943 (2004).

    Article  CAS  Google Scholar 

  20. Raderschall, E. et al. Elevated levels of Rad51 recombination protein in tumor cells. Cancer Res. 62, 219–225 (2002).

    CAS  PubMed  Google Scholar 

  21. Flygare, J., Benson, F. & Hellgren, D. Expression of the human RAD51 gene during the cell cycle in primary human peripheral blood lymphocytes. Biochim. Biophys. Acta 1312, 231–236 (1996).

    Article  Google Scholar 

  22. Yamamoto, A. et al. Cell cycle-dependent expression of the mouse Rad51 gene in proliferating cells. Mol. Gen. Genet. 251, 1–12 (1996).

    CAS  PubMed  Google Scholar 

  23. Chen, F. et al. Cell cycle-dependent protein expression of mammalian homologs of yeast DNA double-strand break repair genes Rad51 and Rad52. Mutat. Res. 384, 205–211 (1997).

    Article  CAS  Google Scholar 

  24. 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 

  25. Moynahan, M.E., Pierce, A.J. & Jasin, M. BRCA2 is required for homology-directed repair of chromosomal breaks. Mol. Cell 7, 263–272 (2001).

    Article  CAS  Google Scholar 

  26. Weaver, D.T. What to do at an end: DNA double-strand-break repair. Trends Genet. 11, 388–392 (1995).

    Article  CAS  Google Scholar 

  27. 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  CAS  Google Scholar 

  28. Takata, M. et al. Homologous recombination and non-homologous end-joining pathways of DNA double-strand break repair have overlapping roles in the maintenance of chromosomal integrity in vertebrate cells. EMBO J. 17, 5497–5508 (1998).

    Article  CAS  Google Scholar 

  29. Johnson, R.D. & Jasin, M. Double-strand-break-induced homologous recombination in mammalian cells. Biochem. Soc. Trans. 29, 196–201 (2001).

    Article  CAS  Google Scholar 

  30. Xu, B., Kim, S.T., Lim, D.S. & Kastan, M.B. Two molecularly distinct G(2)/M checkpoints are induced by ionizing irradiation. Mol. Cell. Biol. 22, 1049–1059 (2002).

    Article  CAS  Google Scholar 

  31. Taniguchi, T. et al. Convergence of the fanconi anemia and ataxia telangiectasia signaling pathways. Cell 109, 459–472 (2002).

    Article  CAS  Google Scholar 

  32. Xie, A. et al. Control of sister chromatid recombination by histone H2AX. Mol. Cell 16, 1017–1025 (2004).

    Article  CAS  Google Scholar 

  33. Miyazaki, T., Bressan, D.A., Shinohara, M., Haber, J.E. & Shinohara, A. In vivo assembly and disassembly of Rad51 and Rad52 complexes during double-strand break repair. EMBO J. 23, 939–949 (2004).

    Article  CAS  Google Scholar 

  34. Bishop, D.K. RecA homologs Dmc1 and Rad51 interact to form multiple nuclear complexes prior to meiotic chromosome synapsis. Cell 79, 1081–1092 (1994).

    Article  CAS  Google Scholar 

  35. Lukas, C. et al. Mdc1 couples DNA double-strand break recognition by Nbs1 with its H2AX-dependent chromatin retention. EMBO J. 23, 2674–2683 (2004).

    Article  CAS  Google Scholar 

  36. Mekeel, K.L. et al. Inactivation of p53 results in high rates of homologous recombination. Oncogene 14, 1847–1857 (1997).

    Article  CAS  Google Scholar 

  37. Xia, F. et al. Deficiency of human BRCA2 leads to impaired homologous recombination but maintains normal nonhomologous end joining. Proc. Natl. Acad. Sci. USA 98, 8644–8649 (2001).

    Article  CAS  Google Scholar 

  38. Chatterjee, A. et al. Mapping the sites of putative tumor suppressor genes at 6p25 and 6p21.3 in cervical carcinoma: occurrence of allelic deletions in precancerous lesions. Cancer Res. 61, 2119–2123 (2001).

    CAS  PubMed  Google Scholar 

  39. Mendez, J. & Stillman, B. Chromatin association of human origin recognition complex, cdc6, and minichromosome maintenance proteins during the cell cycle: assembly of prereplication complexes in late mitosis. Mol. Cell. Biol. 20, 8602–8612 (2000).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank J. Chen and M. Jasin for their generous contribution of materials. This work was supported by Public Health Science grant CA107640 to S.N.P.

Author information

Author notes

  1. Junran Zhang and Zhefu Ma: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Radiation Oncology, MGH Cancer Center, Massachusetts General Hospital and Harvard Medical School, 149 13th Street, Charlestown, 02129, Massachusetts, USA

    Junran Zhang, Alejandro Treszezamsky & Simon N Powell

  2. Division of Gastroenterology, Department of Medicine, Beth Israel Deaconess Medical Center and Harvard Medical School, 99 Brookline Ave., Boston, 02215, Massachusetts, USA

    Zhefu Ma

  3. Department of Radiation Oncology, Washington University School of Medicine, 4511 Forest Park, St. Louis, 63108, Missouri, USA

    Simon N Powell

Authors
  1. Junran Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Zhefu Ma
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Alejandro Treszezamsky
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Simon N Powell
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Simon N Powell.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Zhang, J., Ma, Z., Treszezamsky, A. et al. MDC1 interacts with Rad51 and facilitates homologous recombination. Nat Struct Mol Biol 12, 902–909 (2005). https://doi.org/10.1038/nsmb991

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nsmb991

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing