Abstract
Common fragile sites are specific genomic loci that form constrictions and gaps on metaphase chromosomes under conditions that slow, but do not arrest, DNA replication. These sites have been shown to have a role in various chromosomal rearrangements in tumors. Different DNA damage response proteins were shown to regulate fragile site stability, including ataxia-telangiectasia and Rad3-related (ATR) and its effector Chk1. Here, we investigated the role of ataxia-telangiectasia mutated (ATM), the main transducer of DNA double-strand break (DSB) signal, in this regulation. We demonstrate that replication stress conditions, which induce fragile site expression, lead to DNA fragmentation and recruitment of phosphorylated ATM to nuclear foci at DSBs. We further show that ATM plays a role in maintaining fragile site stability, which is revealed only in the absence of ATR. However, the activation of ATM under these replication stress conditions is ATR independent. Following conditions that induce fragile site expression both ATR and ATM phosphorylate Chk1, suggesting that both proteins regulate fragile site expression probably via their effect on Chk1 activation. Our findings provide new insights into the interplay between ATR and ATM pathways in response to partial replication inhibition and in the regulation of fragile site stability.
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References
Abraham RT . (2001). Cell cycle checkpoint signaling through the ATM and ATR kinases. Genes Dev 15: 2177–2196.
Arlt MF, Casper AM, Glover TW . (2003). Common fragile sites. Cytogenet Genome Res 100: 92–100.
Arlt MF, Durkin SG, Ragland RL, Glover TW . (2006). Common fragile sites as targets for chromosome rearrangements. DNA Repair (Amst) 5: 1126–1135.
Bakkenist CJ, Kastan MB . (2003). DNA damage activates ATM through intermolecular autophosphorylation and dimer dissociation. Nature 421: 499–506.
Bakkenist CJ, Kastan MB . (2004). Initiating cellular stress responses. Cell 118: 9–17.
Banin S, Moyal L, Shieh S, Taya Y, Anderson CW, Chessa L et al. (1998). Enhanced phosphorylation of p53 by ATM in response to DNA damage. Science 281: 1674–1677.
Bartkova J, Horejsi Z, Koed K, Kramer A, Tort F, Zieger K et al (2005). DNA damage response as a candidate anti-cancer barrier in early human tumorigenesis. Nature 434: 864–870.
Bartkova J, Rezaei N, Liontos M, Karakaidos P, Kletsas D, Issaeva N et al. (2006). Oncogene-induced senescence is part of the tumorigenesis barrier imposed by DNA damage checkpoints. Nature 444: 633–637.
Bester AC, Schwartz M, Schmidt M, Garrigue A, Hacein-Bey-Abina S, Cavazzana-Calvo M et al. (2006). Fragile sites are preferential targets for integrations of MLV vectors in gene therapy. Gene Ther 13: 1057–1059.
Bolderson E, Scorah J, Helleday T, Smythe C, Meuth M . (2004). ATM is required for the cellular response to thymidine induced replication fork stress. Hum Mol Genet 13: 2937–2945.
Brown EJ, Baltimore D . (2003). Essential and dispensable roles of ATR in cell cycle arrest and genome maintenance. Genes Dev 17: 615–628.
Canman CE, Lim DS, Cimprich KA, Taya Y, Tamai K, Sakaguchi K et al. (1998). Activation of the ATM kinase by ionizing radiation and phosphorylation of p53. Science 281: 1677–1679.
Casper AM, Nghiem P, Arlt MF, Glover TW . (2002). ATR regulates fragile site stability. Cell 111: 779–789.
Cheng CH, Kuchta RD . (1993). DNA polymerase epsilon: aphidicolin inhibition and the relationship between polymerase and exonuclease activity. Biochemistry 32: 8568–8574.
Cuadrado M, Martinez-Pastor B, Fernandez-Capetillo O . (2006a). ATR activation in response to ionizing radiation: still ATM territory. Cell Div 1: 7.
Cuadrado M, Martinez-Pastor B, Murga M, Toledo LI, Gutierrez-Martinez P, Lopez E et al. (2006b). ATM regulates ATR chromatin loading in response to DNA double-strand breaks. J Exp Med 203: 297–303.
Di Micco R, Fumagalli M, Cicalese A, Piccinin S, Gasparini P, Luise C et al. (2006). Oncogene-induced senescence is a DNA damage response triggered by DNA hyper-replication. Nature 444: 638–642.
Durkin SG, Arlt MF, Howlett NG, Glover TW . (2006). Depletion of CHK1, but not CHK2, induces chromosomal instability and breaks at common fragile sites. Oncogene 25: 4381–4388.
Gatei M, Zhou BB, Hobson K, Scott S, Young D, Khanna KK . (2001). Ataxia telangiectasia mutated (ATM) kinase and ATM and Rad3 related kinase mediate phosphorylation of Brca1 at distinct and overlapping sites. In vivo assessment using phospho-specific antibodies. J Biol Chem 276: 17276–17280.
Glover TW, Stein CK . (1987). Induction of sister chromatid exchanges at common fragile sites. Am J Hum Genet 41: 882–890.
Glover TW, Stein CK . (1988). Chromosome breakage and recombination at fragile sites. Am J Hum Genet 43: 265–273.
Gorgoulis VG, Vassiliou LV, Karakaidos P, Zacharatos P, Kotsinas A, Liloglou T et al. (2005). Activation of the DNA damage checkpoint and genomic instability in human precancerous lesions. Nature 434: 907–913.
Hecht F, Hecht BK . (1984). Fragile sites and chromosome breakpoints in constitutional rearrangements I. Amniocentesis. Clin Genet 26: 169–173.
Hellman A, Rahat A, Scherer SW, Darvasi A, Tsui LC, Kerem B . (2000). Replication delay along FRA7 H, a common fragile site on human chromosome 7, leads to chromosomal instability. Mol Cell Biol 20: 4420–4427.
Ho GP, Margossian S, Taniguchi T, D'Andrea AD . (2006). Phosphorylation of FANCD2 on two novel sites is required for mitomycin C resistance. Mol Cell Biol 26: 7005–7015.
Ikegami S, Taguchi T, Ohashi M, Oguro M, Nagano H, Mano Y . (1978). Aphidicolin prevents mitotic cell division by interfering with the activity of DNA polymerase-alpha. Nature 275: 458–460.
Jazayeri A, Falck J, Lukas C, Bartek J, Smith GC, Lukas J et al. (2006). ATM- and cell cycle-dependent regulation of ATR in response to DNA double-strand breaks. Nat Cell Biol 8: 37–45.
Kim ST, Xu B, Kastan MB . (2002). Involvement of the cohesin protein, SMC1, in ATM-dependent and independent responses to DNA damage. Genes Dev 16: 560–570.
Le Beau MM, Rassool FV, Neilly ME, Espinosa III R, Glover TW, Smith DI et al. (1998). Replication of a common fragile site, FRA3B, occurs late in S phase and is delayed further upon induction: implications for the mechanism of fragile site induction. Hum Mol Genet 7: 755–761.
Lichter P, Cremer T, Borden J, Manuelidis L, Ward DC . (1988). Delineation of individual human chromosomes in metaphase and interphase cells by in situ suppression hybridization using recombinant DNA libraries. Hum Genet 80: 224–234.
Lopes M, Cotta-Ramusino C, Pellicioli A, Liberi G, Plevani P, Muzi-Falconi M et al. (2001). The DNA replication checkpoint response stabilizes stalled replication forks. Nature 412: 557–561.
Myers JS, Cortez D . (2006). Rapid activation of ATR by ionizing radiation requires ATM and Mre11. J Biol Chem 281: 9346–9350.
Richards RI . (2001). Fragile and unstable chromosomes in cancer: causes and consequences. Trends Genet 17: 339–345.
Rogakou EP, Boon C, Redon C, Bonner WM . (1999). Megabase chromatin domains involved in DNA double-strand breaks in vivo. J Cell Biol 146: 905–916.
Rogakou EP, Pilch DR, Orr AH, Ivanova VS, Bonner WM . (1998). DNA double-stranded breaks induce histone H2AX phosphorylation on serine 139. J Biol Chem 273: 5858–5868.
Schwartz M, Zlotorynski E, Goldberg M, Ozeri E, Rahat A, le Sage C et al. (2005). Homologous recombination and nonhomologous end-joining repair pathways regulate fragile site stability. Genes Dev 19: 2715–2726.
Schwartz M, Zlotorynski E, Kerem B . (2006). The molecular basis of common and rare fragile sites. Cancer Lett 232: 13–26.
Shiloh Y . (2006). The ATM-mediated DNA-damage response: taking shape. Trends Biochem Sci 31: 402–410.
Stiff T, Walker SA, Cerosaletti K, Goodarzi AA, Petermann E, Concannon P et al. (2006). ATR-dependent phosphorylation and activation of ATM in response to UV treatment or replication fork stalling. Embo J 25: 5775–5782.
Tercero JA, Diffley JF . (2001). Regulation of DNA replication fork progression through damaged DNA by the Mec1/Rad53 checkpoint. Nature 412: 553–557.
Thorland EC, Myers SL, Gostout BS, Smith DI . (2003). Common fragile sites are preferential targets for HPV16 integrations in cervical tumors. Oncogene 22: 1225–1237.
Wang L, Paradee W, Mullins C, Shridhar R, Rosati R, Wilke CM et al. (1997). Aphidicolin-induced FRA3B breakpoints cluster in two distinct regions. Genomics 41: 485–488.
Yoo HY, Kumagai A, Shevchenko A, Dunphy WG . (2007). Ataxia-telangiectasia mutated (ATM)-dependent activation of ATR occurs through phosphorylation of TopBP1 by ATM. J Biol Chem 282: 17501–17506.
Yunis JJ, Soreng A . (1984). Constitutive fragile sites and cancer. Science 226: 1199–1204.
Ziv Y, Bar-Shira A, Pecker I, Russell P, Jorgensen TJ, Tsarfati I et al. (1997). Recombinant ATM protein complements the cellular A-T phenotype. Oncogene 15: 159–167.
Zou L, Elledge SJ . (2003). Sensing DNA damage through ATRIP recognition of RPA-ssDNA complexes. Science 300: 1542–1548.
Acknowledgements
We thank Naomi Melamed-Book for assistance in confocal analyses, Yifat Eliezer for assistance in western blot analyses, Y Shiloh and Y Ziv for comments on the manuscript, the AT fibroblast cell line (AT22IJE-T) and ATM complemented cells and for the ATM antibody.
This research was partially supported by grants from the Ministry of Science and Technology Israel, the Deutsches Krebsforschungszetrum (DKFZ) and the Israel Cancer Association through the donation from Linda R Kaminow in honor of Ed Fox to BK
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Supplementary Information accompanies the paper on the Oncogene website (http://www.nature.com/onc).
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Ozeri-Galai, E., Schwartz, M., Rahat, A. et al. Interplay between ATM and ATR in the regulation of common fragile site stability. Oncogene 27, 2109–2117 (2008). https://doi.org/10.1038/sj.onc.1210849
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DOI: https://doi.org/10.1038/sj.onc.1210849
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