Abstract
The ATM kinase is a tumour suppressor and a key activator of genome integrity checkpoints in mammalian cells exposed to ionizing radiation (IR) and other insults that elicit DNA double-strand breaks (DSBs). In response to IR, autophosphorylation on serine 1981 causes dissociation of ATM dimers and initiates cellular ATM kinase activity. Here, we show that the kinetics and magnitude of ATM Ser1981 phosphorylation after exposure of human fibroblasts to low doses (2 Gy) of IR are altered in cells deficient in Nbs1, a substrate of ATM and a component of the MRN (Mre11–Rad50–Nbs1) complex involved in processing/repair of DSBs and ATM-dependent cell cycle checkpoints. Timely phosphorylation of both ATM Ser1981 and the ATM substrate Smc1 after IR were rescued via retrovirally mediated reconstitution of Nbs1-deficient cells by wild-type Nbs1 or mutants of Nbs1 defective in the FHA domain or nonphosphorylatable by ATM, but not by Nbs1 lacking the Mre11-interaction domain. Our data indicate that apart from its role downstream of ATM in the DNA damage checkpoint network, the MRN complex serves also as a modulator/amplifier of ATM activity. Although not absolutely required for ATM activation, the MRN nuclease complex may help reach the threshold activity of ATM necessary for optimal genome maintenance and prevention of cancer.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 50 print issues and online access
$259.00 per year
only $5.18 per issue
Rent or buy this article
Prices vary by article type
from$1.95
to$39.95
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Ahn JY, Lee KS, Choi SY and Cho SW . (2000). Cancer Res., 60, 5934–5936.
Anderson L, Henderson C and Adachi Y . (2001). Mol. Cell. Biol., 21, 1719–1729.
Bakkenist CJ and Kastan MB . (2003). Nature, 421, 486–488.
Buscemi G, Savio C, Zannini L, Micciche F, Masnada D, Nakanishi M, Tauchi H, Komatsu K, Mizutani S, Khanna K, Chen P, Concannon P, Chessa L and Delia D . (2001). Mol. Cell. Biol., 21, 5214–5222.
Celeste A, Fernandez-Capetillo O, Kruhlak MJ, Pilch DR, Staudt DW, Lee A, Bonner RF, Bonner WM and Nussenzweig A . (2003). Nat. Cell Biol., 5, 675–679.
Cerosaletti KM and Concannon P . (2003). J. Biol. Chem., 278, 21944–21951.
D'Amours D and Jackson SP . (2002). Nat. Rev. Mol. Cell. Biol., 3, 317–327.
Desai-Mehta A, Cerosaletti KM and Concannon P . (2001). Mol. Cell. Biol., 21, 2184–2191.
Dong Z, Zhong Q and Chen PL . (1999). J. Biol. Chem., 274, 19513–19516.
Durocher D and Jackson SP . (2001). Curr. Opin. Cell Biol., 13, 225–231.
Falck J, Petrini JH, Williams BR, Lukas J and Bartek J . (2002). Nat. Genet., 30, 290–294.
Gatei M, Sloper K, Sφrensen C, Syljuasen R, Falck J, Hobson K, Savage K, Lukas J, Zhou BB, Bartek J and Khanna KK . (2003). J. Biol. Chem., 278, 14806–14811.
Gatei M, Young D, Cerosaletti KM, Desai-Mehta A, Spring K, Kozlov S, Lavin MF, Gatti RA, Concannon P and Khanna K . (2000). Nat. Genet., 25, 115–119.
Gatei M, Zhou BB, Hobson K, Scott S, Young D and Khanna KK . (2001). J. Biol. Chem., 276, 17276–17280.
Girard PM, Riballo E, Begg AC, Waugh A and Jeggo PA . (2002). Oncogene, 21, 4191–4199.
Kastan MB and Lim DS . (2000). Nat. Rev. Mol. Cell. Biol., 1, 179–186.
Khanna KK and Jackson SP . (2001). Nat. Genet., 27, 247–254.
Kim ST, Lim DS, Canman CE and Kastan MB . (1999). J. Biol. Chem., 274, 37538–37543.
Kim ST, Xu B and Kastan MB . (2002). Genes Dev., 16, 560–570.
Lee JH, Xu B, Lee CH, Ahn JY, Song MS, Lee H, Canman CE, Lee JS, Kastan MB and Lim DS . (2003). Mol. Cancer Res., 1, 674–681.
Lim DS, Kim ST, Xu B, Maser RS, Lin J, Petrini JH and Kastan MB . (2000). Nature, 404, 613–617.
Lou Z, Minter-Dykhouse K, Wu X and Chen J . (2003). Nature, 421, 957–961.
Lukas C, Falck J, Bartkova J, Bartek J and Lukas J . (2003). Nat. Cell Biol., 5, 255–260.
Lukas J, Herzinger T, Hansen K, Moroni MC, Resnitzky D, Helin K, Reed SI and Bartek J . (1997). Genes Dev., 11, 1479–1492.
Matsuoka S, Huang M and Elledge SJ . (2000). Proc. Natl. Acad. Sci. USA, 97, 10389–10394.
Mirzoeva OK and Petrini JH . (2001). Mol. Cell. Biol., 21, 281–288.
Okada S and Ouchi T . (2003). J. Biol. Chem., 278, 2015–2020.
Pear WS, Nolan GP, Scott ML and Baltimore D . (1993). Proc. Natl. Acad. Sci. USA, 90, 8392–8396.
Petrini JH and Stracker TH . (2003). Trends Cell Biol., 13, 458–462.
Petrini JH . (2000). Curr. Opin. Cell Biol., 12, 293–296.
Shiloh Y . (1997). Annu. Rev. Genet., 31, 635–662.
Shiloh Y . (2001). Curr. Opin. Genet. Dev., 11, 71–77.
Shiloh Y . (2003). Nat. Rev. Cancer, 3, 155–168.
Sørensen CS, Lukas C, Kramer ER, Peters JM, Bartek J and Lukas J . (2000). Mol. Cell. Biol., 20, 7613–7623.
Sørensen CS, Syljuasen RG, Falck J, Schroeder T, Ronnstrand L, Khanna KK, Zhou BB, Bartek J and Lukas J . (2003). Cancer Cell, 3, 247–258.
Stewart GS, Maser RS, Stankovic T, Bressan DA, Kaplan MI, Jaspers NG, Raams A, Byrd PJ, Petrini JH and Taylor AM . (1999). Cell, 99, 577–587.
Thompson LH and Schild D . (2002). Mutat. Res., 509, 49–78.
Uziel T, Lerenthal Y, Moyal L, Andegeko Y, Mittelman L and Shiloh Y . (2003). EMBO J., 22, 5612–5621.
Varon R, Vissinga C, Platzer M, Cerosaletti KM, Chrzanowska KH, Saar K, Beckmann G, Seemanova E, Cooper PR, Nowak NJ, Stumm M, Weemaes CM, Gatti RA, Wilson RK, Digweed M, Rosenthal A, Sperling K, Concannon P and Reis A . (1998). Cell, 93, 467–476.
Yazdi PT, Wang Y, Zhao S, Patel N, Lee EY and Qin J . (2002). Genes Dev., 16, 571–582.
Zhao S, Renthal W and Lee EY . (2002). Nucleic Acids Res., 30, 4815–4822.
Acknowledgements
We acknowledge support from the Danish Cancer Society, the European Union, NIH (grants CA 71387 and CA 21765), the American Lebanese Syrian Charities of the St. Jude Children's Research Hospital and the Danish Medical Research Council.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Hořejší, Z., Falck, J., Bakkenist, C. et al. Distinct functional domains of Nbs1 modulate the timing and magnitude of ATM activation after low doses of ionizing radiation. Oncogene 23, 3122–3127 (2004). https://doi.org/10.1038/sj.onc.1207447
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/sj.onc.1207447
Keywords
This article is cited by
-
Radio-selective effects of a natural occurring muscle-derived dipeptide in A549 and normal cell lines
Scientific Reports (2019)
-
NBN Phosphorylation regulates the accumulation of MRN and ATM at sites of DNA double-strand breaks
Oncogene (2013)
-
FOXO3 signalling links ATM to the p53 apoptotic pathway following DNA damage
Nature Communications (2012)
-
Ionizing radiation induced signaling of DNA damage response molecules in RAW 264.7 and CD4+ T cells
Molecular and Cellular Biochemistry (2012)
-
The p53-inducible gene 3 (PIG3) contributes to early cellular response to DNA damage
Oncogene (2010)