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

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. Access through your institution Buy or subscribe This is a preview of subscription content, access via your institution ACCESS OPTIONS

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institutional subscriptions * Read our FAQs * Contact customer support SIMILAR CONTENT BEING VIEWED BY OTHERS 3D GENOME ORGANIZATION CONTRIBUTES TO GENOME INSTABILITY AT FRAGILE SITES

Article Open access 17 July 2020 MISTIMED ORIGIN LICENSING AND ACTIVATION STABILIZE COMMON FRAGILE SITES UNDER TIGHT DNA-REPLICATION CHECKPOINT ACTIVATION Article 06 April 2023 FANCD2

MODULATES THE MITOCHONDRIAL STRESS RESPONSE TO PREVENT COMMON FRAGILE SITE INSTABILITY Article Open access 29 January 2021 REFERENCES * Abraham RT . (2001). Cell cycle checkpoint signaling

through the ATM and ATR kinases. _Genes Dev_ 15: 2177–2196. Article  CAS  PubMed  Google Scholar  * Arlt MF, Casper AM, Glover TW . (2003). Common fragile sites. _Cytogenet Genome Res_ 100:

92–100. Article  CAS  PubMed  Google Scholar  * Arlt MF, Durkin SG, Ragland RL, Glover TW . (2006). Common fragile sites as targets for chromosome rearrangements. _DNA Repair (Amst)_ 5:

1126–1135. Article  CAS  Google Scholar  * Bakkenist CJ, Kastan MB . (2003). DNA damage activates ATM through intermolecular autophosphorylation and dimer dissociation. _Nature_ 421:

499–506. Article  CAS  PubMed  Google Scholar  * Bakkenist CJ, Kastan MB . (2004). Initiating cellular stress responses. _Cell_ 118: 9–17. Article  CAS  PubMed  Google Scholar  * 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. Article  CAS  PubMed  Google

Scholar  * 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. CAS  PubMed  Google Scholar  * 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. CAS  PubMed  Google Scholar  * 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. Article  CAS  PubMed  Google Scholar  * 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. Article  CAS  PubMed  Google Scholar  *

Brown EJ, Baltimore D . (2003). Essential and dispensable roles of ATR in cell cycle arrest and genome maintenance. _Genes Dev_ 17: 615–628. Article  CAS  PubMed  PubMed Central  Google

Scholar  * 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. Article  CAS  PubMed  Google Scholar  * Casper AM, Nghiem P, Arlt MF, Glover TW . (2002). ATR regulates fragile site stability. _Cell_ 111: 779–789. Article  CAS  PubMed  Google

Scholar  * Cheng CH, Kuchta RD . (1993). DNA polymerase epsilon: aphidicolin inhibition and the relationship between polymerase and exonuclease activity. _Biochemistry_ 32: 8568–8574.

Article  CAS  PubMed  Google Scholar  * Cuadrado M, Martinez-Pastor B, Fernandez-Capetillo O . (2006a). ATR activation in response to ionizing radiation: still ATM territory. _Cell Div_ 1:

7. Article  PubMed  PubMed Central  Google Scholar  * 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. Article  CAS  PubMed  PubMed Central  Google Scholar  * 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. Article  CAS  PubMed  Google Scholar  * 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. Article  CAS  PubMed 

Google Scholar  * 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. Article  CAS  PubMed  Google Scholar  * Glover TW, Stein CK .

(1987). Induction of sister chromatid exchanges at common fragile sites. _Am J Hum Genet_ 41: 882–890. CAS  PubMed  PubMed Central  Google Scholar  * Glover TW, Stein CK . (1988).

Chromosome breakage and recombination at fragile sites. _Am J Hum Genet_ 43: 265–273. CAS  PubMed  PubMed Central  Google Scholar  * 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. Article  CAS  PubMed  Google

Scholar  * Hecht F, Hecht BK . (1984). Fragile sites and chromosome breakpoints in constitutional rearrangements I. Amniocentesis. _Clin Genet_ 26: 169–173. Article  CAS  PubMed  Google

Scholar  * 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. Article  CAS  PubMed  PubMed Central  Google Scholar  * 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. Article  CAS  PubMed  PubMed Central  Google Scholar  * 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. Article  CAS  PubMed  Google Scholar  *

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.

Article  CAS  PubMed  Google Scholar  * 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. Article  CAS  PubMed  PubMed Central  Google Scholar  * 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. Article  CAS  PubMed  Google

Scholar  * 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. Article  CAS  PubMed  Google Scholar  * 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. Article  CAS  PubMed  Google Scholar  * Myers JS, Cortez D . (2006). Rapid

activation of ATR by ionizing radiation requires ATM and Mre11. _J Biol Chem_ 281: 9346–9350. Article  CAS  PubMed  Google Scholar  * Richards RI . (2001). Fragile and unstable chromosomes

in cancer: causes and consequences. _Trends Genet_ 17: 339–345. Article  CAS  PubMed  Google Scholar  * 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. Article  CAS  PubMed  PubMed Central  Google Scholar  * 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. Article  CAS  PubMed  Google Scholar  * 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. Article  CAS  PubMed

  PubMed Central  Google Scholar  * Schwartz M, Zlotorynski E, Kerem B . (2006). The molecular basis of common and rare fragile sites. _Cancer Lett_ 232: 13–26. Article  CAS  PubMed  Google

Scholar  * Shiloh Y . (2006). The ATM-mediated DNA-damage response: taking shape. _Trends Biochem Sci_ 31: 402–410. Article  CAS  PubMed  Google Scholar  * 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.

Article  CAS  PubMed  PubMed Central  Google Scholar  * Tercero JA, Diffley JF . (2001). Regulation of DNA replication fork progression through damaged DNA by the Mec1/Rad53 checkpoint.

_Nature_ 412: 553–557. Article  CAS  PubMed  Google Scholar  * 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. Article  CAS  PubMed  Google Scholar  * 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. Article  CAS  PubMed  Google Scholar  * 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. Article  CAS  PubMed  Google Scholar  * Yunis JJ, Soreng A .

(1984). Constitutive fragile sites and cancer. _Science_ 226: 1199–1204. Article  CAS  PubMed  Google Scholar  * 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. Article  CAS  PubMed  Google Scholar  * Zou L, Elledge SJ . (2003). Sensing DNA damage through

ATRIP recognition of RPA-ssDNA complexes. _Science_ 300: 1542–1548. Article  CAS  PubMed  Google Scholar  Download references 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 AUTHOR INFORMATION AUTHORS AND AFFILIATIONS * Department of Genetics, The Life Sciences Institute, The

Hebrew University, Jerusalem, Israel E Ozeri-Galai, M Schwartz, A Rahat & B Kerem Authors * E Ozeri-Galai View author publications You can also search for this author inPubMed Google

Scholar * M Schwartz View author publications You can also search for this author inPubMed Google Scholar * A Rahat View author publications You can also search for this author inPubMed 

Google Scholar * B Kerem View author publications You can also search for this author inPubMed Google Scholar CORRESPONDING AUTHOR Correspondence to B Kerem. ADDITIONAL INFORMATION

Supplementary Information accompanies the paper on the Oncogene website (http://www.nature.com/onc). SUPPLEMENTARY INFORMATION SUPPLEMENTARY INFORMATION (DOC 20 KB) SUPPLEMENTARY FIGURE S1

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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

Download citation * Received: 16 July 2007 * Revised: 12 September 2007 * Accepted: 13 September 2007 * Published: 15 October 2007 * Issue Date: 03 April 2008 * DOI:

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currently available for this article. Copy to clipboard Provided by the Springer Nature SharedIt content-sharing initiative KEYWORDS * fragile sites * replication stress * ATM * ATR * double

strand breaks * Chk1