DNA Damage Sensing and Signaling

  • Daniel DurocherEmail author


In order to maintain genome integrity, cells have evolved a complex network of processes that detect, repair and signal DNA damage. In this chapter, I review the early steps of DNA damage signaling with a particular emphasis on how DNA lesions are sensed and how the detection of DNA damage leads to the activation of DNA damage signaling by the ATM and ATR protein kinases.


ATM ATR MRN DNA damage signaling Checkpoint 



I am indebted to Rachel Szilard who has patiently proofread this manuscript. The ideas on the bypass of the loss of ATM by activating DNA-PK were formed during discussions with Steve Jackson and Andre Nussenzweig. Finally, work on DNA damage signaling in my laboratory is supported by grants from the Canadian Institutes of Health Research (MOP89754 and MOP84297).


  1. 1.
    Friedberg EC (2003) DNA damage and repair. Nature 421: 436–440.PubMedGoogle Scholar
  2. 2.
    Fraser CM, Gocayne JD, White O, Adams MD, Clayton RA, Fleischmann RD, Bult CJ, Kerlavage AR, Sutton G, Kelley JM, Fritchman RD, Weidman JF, Small KV, Sandusky M, Fuhrmann J, Nguyen D, Utterback TR, Saudek DM, Phillips CA, Merrick JM, Tomb JF, Dougherty BA, Bott KF, Hu PC, Lucier TS, Peterson SN, Smith HO, Hutchison CA, 3rd, and Venter JC (1995) The minimal gene complement of Mycoplasma genitalium. Science 270: 397–403.PubMedGoogle Scholar
  3. 3.
    Waters E, Hohn MJ, Ahel I, Graham DE, Adams MD, Barnstead M, Beeson KY, Bibbs L, Bolanos R, Keller M, Kretz K, Lin X, Mathur E, Ni J, Podar M, Richardson T, Sutton GG, Simon M, Soll D, Stetter KO, Short JM, and Noordewier M (2003) The genome of Nanoarchaeum equitans: insights into early archaeal evolution and derived parasitism. Proc Natl Acad Sci USA 100: 12984–12988.PubMedGoogle Scholar
  4. 4.
    Painter RB, and Young BR (1980) Radiosensitivity in ataxia-telangiectasia: a new explanation. Proc Natl Acad Sci USA 77: 7315–7317.PubMedGoogle Scholar
  5. 5.
    Weinert TA, and Hartwell LH (1988) The RAD9 gene controls the cell cycle response to DNA damage in Saccharomyces cerevisiae. Science 241: 317–322.PubMedGoogle Scholar
  6. 6.
    Hartwell LH, and Weinert TA (1989) Checkpoints: controls that ensure the order of cell cycle events. Science 246: 629–634.PubMedGoogle Scholar
  7. 7.
    Hartley KO, Gell D, Smith GC, Zhang H, Divecha N, Connelly MA, Admon A, Lees-Miller SP, Anderson CW, and Jackson SP (1995) DNA-dependent protein kinase catalytic subunit: a relative of phosphatidylinositol 3-kinase and the ataxia telangiectasia gene product. Cell 82: 849–856.PubMedGoogle Scholar
  8. 8.
    Jackson SP, MacDonald JJ, Lees-Miller S, and Tjian R (1990) GC box binding induces phosphorylation of Sp1 by a DNA-dependent protein kinase. Cell 63: 155–165.PubMedGoogle Scholar
  9. 9.
    Lees-Miller SP, and Anderson CW (1989) The human double-stranded DNA-activated protein kinase phosphorylates the 90-kDa heat-shock protein, hsp90 alpha at two NH2-terminal threonine residues. J Biol Chem 264: 17275–17280.PubMedGoogle Scholar
  10. 10.
    Sun Z, Fay DS, Marini F, Foiani M, and Stern DF (1996) Spk1/Rad53 is regulated by Mec1-dependent protein phosphorylation in DNA replication and damage checkpoint pathways. Genes Dev 10: 395–406.PubMedGoogle Scholar
  11. 11.
    Savitsky K, Barshira A, Gilad S, Rotman G, Ziv Y, Vanagaite L, Tagle DA, Smith S, Uziel T, Sfez S, Ashkenazi M, Pecker I, Frydman M, Harnik R, Patanjali SR, Simmons A, Clines GA, Sartiel A, Gatti RA, Chessa L, Sanal O, Lavin MF, Jaspers NGJ, Malcolm A, Taylor R, Arlett CF, Miki T, Weissman SM, Lovett M, Collins FS, and Shiloh Y (1995) A single ataxia telangiectasia gene with a product similar to Pi 3 kinase. Science 268: 1749–1753.PubMedGoogle Scholar
  12. 12.
    Greenwell PW, Kronmal SL, Porter SE, Gassenhuber J, Obermaier B, and Petes TD (1995) TEL1, a gene involved in controlling telomere length in S. cerevisiae, is homologous to the human ataxia telangiectasia gene. Cell 82: 823–829.PubMedGoogle Scholar
  13. 13.
    Kato R, and Ogawa H (1994) An essential gene, ESR1, is required for mitotic cell growth, DNA repair and meiotic recombination in Saccharomyces cerevisiae. Nucleic Acids Res 22: 3104–3112.PubMedGoogle Scholar
  14. 14.
    Bentley NJ, Holtzman DA, Flaggs G, Keegan KS, DeMaggio A, Ford JC, Hoekstra M, and Carr AM (1996) The Schizosaccharomyces pombe rad3 checkpoint gene. Embo J 15: 6641–6651.PubMedGoogle Scholar
  15. 15.
    Matsuoka S, Huang M, and Elledge SJ (1998) Linkage of ATM to cell cycle regulation by the Chk2 protein kinase. Science 282: 1893–1897.PubMedGoogle Scholar
  16. 16.
    Walworth N, Davey S, and Beach D (1993) Fission yeast Chk1 protein kinase links the rad checkpoint pathway to Cdc2. Nature 363: 368–371.PubMedGoogle Scholar
  17. 17.
    Boddy MN, Furnari B, Mondesert O, and Russell P (1998) Replication checkpoint enforced by kinases Cds1 and Chk1. Science 280: 909–912.PubMedGoogle Scholar
  18. 18.
    Allen JB, Zhou Z, Siede W, Friedberg EC, and Elledge SJ (1994) The SAD1/RAD53 protein kinase controls multiple checkpoints and DNA damage-induced transcription in yeast. Genes Dev 8: 2401–2415.PubMedGoogle Scholar
  19. 19.
    Hari KL, Santerre A, Sekelsky JJ, McKim KS, Boyd JB, and Hawley RS (1995) The mei-41 gene of D. melanogaster is a structural and functional homolog of the human ataxia telangiectasia gene. Cell 82: 815–821.PubMedGoogle Scholar
  20. 20.
    Durocher D, and Jackson SP (2001) DNA-PK, ATM and ATR as sensors of DNA damage: variations on a theme? Curr Opin Cell Biol 13: 225–231.PubMedGoogle Scholar
  21. 21.
    Yamashita A, Ohnishi T, Kashima I, Taya Y, and Ohno S (2001) Human SMG-1, a novel phosphatidylinositol 3-kinase-related protein kinase, associates with components of the mRNA surveillance complex and is involved in the regulation of nonsense-mediated mRNA decay. Genes Dev 15: 2215–2228.PubMedGoogle Scholar
  22. 22.
    Brumbaugh KM, Otterness DM, Geisen C, Oliveira V, Brognard J, Li X, Lejeune F, Tibbetts RS, Maquat LE, and Abraham RT (2004) The mRNA surveillance protein hSMG-1 functions in genotoxic stress response pathways in mammalian cells. Mol Cell 14: 585–598.PubMedGoogle Scholar
  23. 23.
    Bannister AJ, Gottlieb TM, Kouzarides T, and Jackson SP (1993) c-Jun is phosphorylated by the DNA-dependent protein kinase in vitro; definition of the minimal kinase recognition motif. Nucleic Acids Res 21: 1289–1295.PubMedGoogle Scholar
  24. 24.
    O'Neill T, Dwyer AJ, Ziv Y, Chan DW, Lees-Miller SP, Abraham RH, Lai JH, Hill D, Shiloh Y, Cantley LC, and Rathbun GA (2000) Utilization of oriented peptide libraries to identify substrate motifs selected by ATM. J Biol Chem 275: 22719–22727.PubMedGoogle Scholar
  25. 25.
    Kim ST, Lim DS, Canman CE, and Kastan MB (1999) Substrate specificities and identification of putative substrates of ATM kinase family members. J Biol Chem 274: 37538–37543.PubMedGoogle Scholar
  26. 26.
    You Z, Bailis JM, Johnson SA, Dilworth SM, and Hunter T (2007) Rapid activation of ATM on DNA flanking double-strand breaks. Nat Cell Biol 9: 1311–1318.PubMedGoogle Scholar
  27. 27.
    Lee JH, and Paull TT (2005) ATM activation by DNA double-strand breaks through the Mre11-Rad50-Nbs1 complex. Science 308: 551–554.PubMedGoogle Scholar
  28. 28.
    Smith GC, Cary RB, Lakin ND, Hann BC, Teo SH, Chen DJ, and Jackson SP (1999) Purification and DNA binding properties of the ataxia-telangiectasia gene product ATM. Proc Natl Acad Sci USA 96: 11134–11139.PubMedGoogle Scholar
  29. 29.
    MacDougall CA, Byun TS, Van C, Yee MC, and Cimprich KA (2007) The structural determinants of checkpoint activation. Genes Dev 21: 898–903.PubMedGoogle Scholar
  30. 30.
    Hall-Jackson CA, Cross DA, Morrice N, and Smythe C (1999) ATR is a caffeine-sensitive, DNA-activated protein kinase with a substrate specificity distinct from DNA-PK. Oncogene 18: 6707–6713.PubMedGoogle Scholar
  31. 31.
    Hefferin ML, and Tomkinson AE (2005) Mechanism of DNA double-strand break repair by non-homologous end joining. DNA Repair (Amst) 4: 639–648.Google Scholar
  32. 32.
    Rouse J, and Jackson SP (2002) Interfaces between the detection, signaling, and repair of DNA damage. Science 297: 547–551.PubMedGoogle Scholar
  33. 33.
    Zhou BB, and Elledge SJ (2000) The DNA damage response: putting checkpoints in perspective. Nature 408: 433–439.PubMedGoogle Scholar
  34. 34.
    Matsuoka S, Ballif BA, Smogorzewska A, McDonald ER, 3rd, Hurov KE, Luo J, Bakalarski CE, Zhao Z, Solimini N, Lerenthal Y, Shiloh Y, Gygi SP, and Elledge SJ (2007) ATM and ATR substrate analysis reveals extensive protein networks responsive to DNA damage. Science 316: 1160–1166.PubMedGoogle Scholar
  35. 35.
    Stokes MP, Rush J, Macneill J, Ren JM, Sprott K, Nardone J, Yang V, Beausoleil SA, Gygi SP, Livingstone M, Zhang H, Polakiewicz RD, and Comb MJ (2007) Profiling of UV-induced ATM/ATR signaling pathways. Proc Natl Acad Sci USA 104: 19855–19860.PubMedGoogle Scholar
  36. 36.
    Smolka MB, Albuquerque CP, Chen SH, and Zhou H (2007) Proteome-wide identification of in vivo targets of DNA damage checkpoint kinases. Proc Natl Acad Sci USA 104: 10364–10369.PubMedGoogle Scholar
  37. 37.
    Mimori T, and Hardin JA (1986) Mechanism of interaction between Ku protein and DNA. J Biol Chem 261: 10375–10379.PubMedGoogle Scholar
  38. 38.
    Dynan WS, and Yoo S (1998) Interaction of Ku protein and DNA-dependent protein kinase catalytic subunit with nucleic acids. Nucleic Acids Res 26: 1551–1559.PubMedGoogle Scholar
  39. 39.
    Scharer OD, and Jiricny J (2001) Recent progress in the biology, chemistry and structural biology of DNA glycosylases. Bioessays 23: 270–281.PubMedGoogle Scholar
  40. 40.
    Yoshioka K, Yoshioka Y, and Hsieh P (2006) ATR kinase activation mediated by MutSalpha and MutLalpha in response to cytotoxic O6-methylguanine adducts. Mol Cell 22: 501–510.PubMedGoogle Scholar
  41. 41.
    Stojic L, Mojas N, Cejka P, Di Pietro M, Ferrari S, Marra G, and Jiricny J (2004) Mismatch repair-dependent G2 checkpoint induced by low doses of SN1 type methylating agents requires the ATR kinase. Genes Dev 18: 1331–1344.PubMedGoogle Scholar
  42. 42.
    Sogo JM, Lopes M, and Foiani M (2002) Fork reversal and ssDNA accumulation at stalled replication forks owing to checkpoint defects. Science 297: 599–602.PubMedGoogle Scholar
  43. 43.
    Byun TS, Pacek M, Yee MC, Walter JC, and Cimprich KA (2005) Functional uncoupling of MCM helicase and DNA polymerase activities activates the ATR-dependent checkpoint. Genes Dev 19: 1040–1052.PubMedGoogle Scholar
  44. 44.
    Jeggo PA, and Lobrich M (2007) DNA double-strand breaks: their cellular and clinical impact? Oncogene 26: 7717–7719.PubMedGoogle Scholar
  45. 45.
    Sandell LL, and Zakian VA (1993) Loss of a yeast telomere: arrest, recovery, and chromosome loss. Cell 75: 729–739.PubMedGoogle Scholar
  46. 46.
    Wyman C, and Kanaar R (2006) DNA double-strand break repair: all's well that ends well. Annu Rev Genet 40: 363–383.PubMedGoogle Scholar
  47. 47.
    Lavin MF, and Shiloh Y (1997) The genetic defect in ataxia-telangiectasia. Annu Rev Immunol 15: 177–202.PubMedGoogle Scholar
  48. 48.
    Perry J, and Kleckner N (2003) The ATRs, ATMs, and TORs are giant HEAT repeat proteins. Cell 112: 151–155.PubMedGoogle Scholar
  49. 49.
    Kobe B, Gleichmann T, Horne J, Jennings IG, Scotney PD, and Teh T (1999) Turn up the HEAT. Structure 7: R91–97.PubMedGoogle Scholar
  50. 50.
    Bosotti R, Isacchi A, and Sonnhammer EL (2000) FAT: a novel domain in PIK-related kinases. Trends Biochem Sci 25: 225–227.PubMedGoogle Scholar
  51. 51.
    Bakkenist CJ, and Kastan MB (2003) DNA damage activates ATM through intermolecular autophosphorylation and dimer dissociation. Nature 421: 499–506.PubMedGoogle Scholar
  52. 52.
    Kozlov S, Gueven N, Keating K, Ramsay J, and Lavin MF (2003) ATP activates ataxia-telangiectasia mutated (ATM) in vitro. Importance of autophosphorylation. J Biol Chem 278: 9309–9317.PubMedGoogle Scholar
  53. 53.
    Bekker-Jensen S, Lukas C, Kitagawa R, Melander F, Kastan MB, Bartek J, and Lukas J (2006) Spatial organization of the mammalian genome surveillance machinery in response to DNA strand breaks. J Cell Biol 173: 195–206.PubMedGoogle Scholar
  54. 54.
    Andegeko Y, Moyal L, Mittelman L, Tsarfaty I, Shiloh Y, and Rotman G (2001) Nuclear retention of ATM at sites of DNA double strand breaks. J Biol Chem 276: 38224–38230.PubMedGoogle Scholar
  55. 55.
    Berkovich E, Monnat RJ, Jr., and Kastan MB (2007) Roles of ATM and NBS1 in chromatin structure modulation and DNA double-strand break repair. Nat Cell Biol 9: 683–690.PubMedGoogle Scholar
  56. 56.
    Lisby M, Barlow JH, Burgess RC, and Rothstein R (2004) Choreography of the DNA damage response: spatiotemporal relationships among checkpoint and repair proteins. Cell 118: 699–713.PubMedGoogle Scholar
  57. 57.
    Lukas C, Falck J, Bartkova J, Bartek J, and Lukas J (2003) Distinct spatiotemporal dynamics of mammalian checkpoint regulators induced by DNA damage. Nat Cell Biol 5: 255–260.PubMedGoogle Scholar
  58. 58.
    Banin S, Moyal L, Shieh S, Taya Y, Anderson CW, Chessa L, Smorodinsky NI, Prives C, Reiss Y, Shiloh Y, and Ziv Y (1998) Enhanced phosphorylation of p53 by ATM in response to DNA damage. Science 281: 1674–1677.PubMedGoogle Scholar
  59. 59.
    Canman CE, Lim DS, Cimprich KA, Taya Y, Tamai K, Sakaguchi K, Appella E, Kastan MB, and Siliciano JD (1998) Activation of the ATM kinase by ionizing radiation and phosphorylation of p53. Science 281: 1677–1679.PubMedGoogle Scholar
  60. 60.
    Chan DW, Ye R, Veillette CJ, and Lees-Miller SP (1999) DNA-dependent protein kinase phosphorylation sites in Ku 70/80 heterodimer. Biochemistry 38: 1819–1828.PubMedGoogle Scholar
  61. 61.
    Sweeney FD, Yang F, Chi A, Shabanowitz J, Hunt DF, and Durocher D (2005) Saccharomyces cerevisiae Rad9 Acts as a Mec1 Adaptor to Allow Rad53 Activation. Curr Biol 15: 1364–1375.PubMedGoogle Scholar
  62. 62.
    Kozlov SV, Graham ME, Peng C, Chen P, Robinson PJ, and Lavin MF (2006) Involvement of novel autophosphorylation sites in ATM activation. Embo J 25: 3504–3514.PubMedGoogle Scholar
  63. 63.
    Usui T, Ogawa H, and Petrini JH (2001) A DNA damage response pathway controlled by Tel1 and the Mre11 complex. Mol Cell 7: 1255–1266.PubMedGoogle Scholar
  64. 64.
    You Z, Chahwan C, Bailis J, Hunter T, and Russell P (2005) ATM activation and its recruitment to damaged DNA require binding to the C terminus of Nbs1. Mol Cell Biol 25: 5363–5379.PubMedGoogle Scholar
  65. 65.
    Dupre A, Boyer-Chatenet L, and Gautier J (2006) Two-step activation of ATM by DNA and the Mre11-Rad50-Nbs1 complex. Nat Struct Mol Biol 13: 451–457.PubMedGoogle Scholar
  66. 66.
    Lee JH, and Paull TT (2004) Direct activation of the ATM protein kinase by the Mre11/Rad50/Nbs1 complex. Science 304: 93–96.PubMedGoogle Scholar
  67. 67.
    Stewart GS, Maser RS, Stankovic T, Bressan DA, Kaplan MI, Jaspers NGJ, Raams A, Byrd PJ, Petrini JHJ, and Taylor AMR (1999) The DNA double-strand break repair gene hMRE11 is mutated in individuals with an ataxia-telangiectasia-like disorder. Cell 99: 577–587.PubMedGoogle Scholar
  68. 68.
    Carney JP, Maser RS, Olivares H, Davis EM, LeBeau M, Yates JR, Hays L, Morgan WF, and Petrini JHJ (1998) The hMre11/hRad50 protein complex and Nijmegen breakage syndrome: Linkage of double strand break repair to the cellular DNA damage response. Cell 93: 477–486.PubMedGoogle Scholar
  69. 69.
    Matsuura S, Tauchi H, Nakamura A, Kondo N, Sakamoto S, Endo S, Smeets D, Solder B, Belohradsky BH, Kaloustian VMD, Oshimura M, Isomura M, Nakamura Y, and Komatsu K (1998) Positional cloning of the gene for Nijmegen breakage syndrome. Nature Genetics 19: 179–181.PubMedGoogle Scholar
  70. 70.
    Varon R, Vissinga C, Platzer M, Cerosaletti KM, Chrzanowska KH, Saar K, Beckmann G, Seemanova E, Cooper PR, Nowak NJ, Stumm M, Weemaes CMR, Gatti RA, Wilson RK, Digweed M, Rosenthal A, Sperling K, Concannon P, and Reis A (1998) Nibrin, a novel DNA double-strand break repair protein, is mutated in Nijmegen breakage syndrome. Cell 93: 467–476.PubMedGoogle Scholar
  71. 71.
    Shiloh Y (1997) Ataxia-telangiectasia and the Nijmegen breakage syndrome: related disorders but genes apart. Annu Rev Genet 31: 635–662.PubMedGoogle Scholar
  72. 72.
    Lim DS, Kim ST, Xu B, Maser RS, Lin J, Petrini JH, and Kastan MB (2000) ATM phosphorylates p95/nbs1 in an S-phase checkpoint pathway. Nature 404: 613–617.PubMedGoogle Scholar
  73. 73.
    Wu X, Ranganathan V, Weisman DS, Heine WF, Ciccone DN, O'Neill TB, Crick KE, Pierce KA, Lane WS, Rathbun G, Livingston DM, and Weaver DT (2000) ATM phosphorylation of Nijmegen breakage syndrome protein is required in a DNA damage response. Nature 405: 477–482.PubMedGoogle Scholar
  74. 74.
    Zhao S, Weng YC, Yuan SS, Lin YT, Hsu HC, Lin SC, Gerbino E, Song MH, Zdzienicka MZ, Gatti RA, Shay JW, Ziv Y, Shiloh Y, and Lee EY (2000) Functional link between ataxia-telangiectasia and Nijmegen breakage syndrome gene products. Nature 405: 473–477.PubMedGoogle Scholar
  75. 75.
    D'Amours D, and Jackson SP (2001) The yeast Xrs2 complex functions in S phase checkpoint regulation. Genes Dev 15: 2238–2249.PubMedGoogle Scholar
  76. 76.
    Kitagawa R, Bakkenist CJ, McKinnon PJ, and Kastan MB (2004) Phosphorylation of SMC1 is a critical downstream event in the ATM-NBS1-BRCA1 pathway. Genes Dev 18: 1423–1438.PubMedGoogle Scholar
  77. 77.
    Uziel T, Lerenthal Y, Moyal L, Andegeko Y, Mittelman L, and Shiloh Y (2003) Requirement of the MRN complex for ATM activation by DNA damage. Embo J 22: 5612–5621.PubMedGoogle Scholar
  78. 78.
    Cerosaletti K, and Concannon P (2004) Independent roles for nibrin and Mre11-Rad50 in the activation and function of Atm. J Biol Chem 279: 38813–38819.PubMedGoogle Scholar
  79. 79.
    Difilippantonio S, Celeste A, Fernandez-Capetillo O, Chen HT, Reina San Martin B, Van Laethem F, Yang YP, Petukhova GV, Eckhaus M, Feigenbaum L, Manova K, Kruhlak M, Camerini-Otero RD, Sharan S, Nussenzweig M, and Nussenzweig A (2005) Role of Nbs1 in the activation of the Atm kinase revealed in humanized mouse models. Nat Cell Biol 7: 675–685.PubMedGoogle Scholar
  80. 80.
    Carson CT, Schwartz RA, Stracker TH, Lilley CE, Lee DV, and Weitzman MD (2003) The Mre11 complex is required for ATM activation and the G2/M checkpoint. Embo J 22: 6610–6620.PubMedGoogle Scholar
  81. 81.
    Nakada D, Matsumoto K, and Sugimoto K (2003) ATM-related Tel1 associates with double-strand breaks through an Xrs2-dependent mechanism. Genes Dev 17: 1957–1962.PubMedGoogle Scholar
  82. 82.
    Falck J, Coates J, and Jackson SP (2005) Conserved modes of recruitment of ATM, ATR and DNA-PKcs to sites of DNA damage. Nature 434: 605–611.PubMedGoogle Scholar
  83. 83.
    Stracker TH, Morales M, Couto SS, Hussein H, and Petrini JH (2007) The carboxy terminus of NBS1 is required for induction of apoptosis by the MRE11 complex. Nature 447: 218–221.PubMedGoogle Scholar
  84. 84.
    Difilippantonio S, Celeste A, Kruhlak MJ, Lee Y, Difilippantonio MJ, Feigenbaum L, Jackson SP, McKinnon PJ, and Nussenzweig A (2007) Distinct domains in Nbs1 regulate irradiation-induced checkpoints and apoptosis. J Exp Med 204: 1003–1011.PubMedGoogle Scholar
  85. 85.
    Kanu N, and Behrens A (2007) ATMIN defines an NBS1-independent pathway of ATM signalling. Embo J 26: 2933–2941.PubMedGoogle Scholar
  86. 86.
    Park BJ, Kang JW, Lee SW, Choi SJ, Shin YK, Ahn YH, Choi YH, Choi D, Lee KS, and Kim S (2005) The haploinsufficient tumor suppressor p18 upregulates p53 via interactions with ATM/ATR. Cell 120: 209–221.PubMedGoogle Scholar
  87. 87.
    Sun Y, Jiang X, Chen S, Fernandes N, and Price BD (2005) A role for the Tip60 histone acetyltransferase in the acetylation and activation of ATM. Proc Natl Acad Sci USA 102: 13182–13187.PubMedGoogle Scholar
  88. 88.
    Gupta A, Sharma GG, Young CS, Agarwal M, Smith ER, Paull TT, Lucchesi JC, Khanna KK, Ludwig T, and Pandita TK (2005) Involvement of human MOF in ATM function. Mol Cell Biol 25: 5292–5305.PubMedGoogle Scholar
  89. 89.
    Richard DJ, Bolderson E, Cubeddu L, Wadsworth RI, Savage K, Sharma GG, Nicolette ML, Tsvetanov S, McIlwraith MJ, Pandita RK, Takeda S, Hay RT, Gautier J, West SC, Paull TT, Pandita TK, White MF, and Khanna KK (2008) Single-stranded DNA-binding protein hSSB1 is critical for genomic stability. Nature 453: 677–681.PubMedGoogle Scholar
  90. 90.
    Bhaskara V, Dupre A, Lengsfeld B, Hopkins BB, Chan A, Lee JH, Zhang X, Gautier J, Zakian V, and Paull TT (2007) Rad50 adenylate kinase activity regulates DNA tethering by Mre11/Rad50 complexes. Mol Cell 25: 647–661.PubMedGoogle Scholar
  91. 91.
    Moncalian G, Lengsfeld B, Bhaskara V, Hopfner KP, Karcher A, Alden E, Tainer JA, and Paull TT (2004) The rad50 signature motif: essential to ATP binding and biological function. J Mol Biol 335: 937–951.PubMedGoogle Scholar
  92. 92.
    Lou Z, Minter-Dykhouse K, Franco S, Gostissa M, Rivera MA, Celeste A, Manis JP, van Deursen J, Nussenzweig A, Paull TT, Alt FW, and Chen J (2006) MDC1 maintains genomic stability by participating in the amplification of ATM-dependent DNA damage signals. Mol Cell 21: 187–200.PubMedGoogle Scholar
  93. 93.
    Stucki M, Clapperton JA, Mohammad D, Yaffe MB, Smerdon SJ, and Jackson SP (2005) MDC1 directly binds phosphorylated histone H2AX to regulate cellular responses to DNA double-strand breaks. Cell 123: 1213–1226.PubMedGoogle Scholar
  94. 94.
    Burma S, Chen BP, Murphy M, Kurimasa A, and Chen DJ (2001) ATM phosphorylates histone H2AX in response to DNA double-strand breaks. J Biol Chem 276: 42462–42467.PubMedGoogle Scholar
  95. 95.
    Stiff T, O'Driscoll M, Rief N, Iwabuchi K, Lobrich M, and Jeggo PA (2004) ATM and DNA-PK function redundantly to phosphorylate H2AX after exposure to ionizing radiation. Cancer Res 64: 2390–2396.PubMedGoogle Scholar
  96. 96.
    Paull TT, Rogakou EP, Yamazaki V, Kirchgessner CU, Gellert M, and Bonner WM (2000) A critical role for histone H2AX in recruitment of repair factors to nuclear foci after DNA damage. Curr Biol 10: 886–895.PubMedGoogle Scholar
  97. 97.
    Stucki M, and Jackson SP (2006) gammaH2AX and MDC1: Anchoring the DNA-damage-response machinery to broken chromosomes. DNA Repair (Amst).Google Scholar
  98. 98.
    Soutoglou E, and Misteli T (2008) Activation of the cellular DNA damage response in the absence of DNA lesions. Science 320: 1507–1510.PubMedGoogle Scholar
  99. 99.
    Hammarsten O, and Chu G (1998) DNA-dependent protein kinase: DNA binding and activation in the absence of Ku. Proc Natl Acad Sci USA 95: 525–530.PubMedGoogle Scholar
  100. 100.
    Kim DH, Sarbassov DD, Ali SM, Latek RR, Guntur KV, Erdjument-Bromage H, Tempst P, and Sabatini DM (2003) GbetaL, a positive regulator of the rapamycin-sensitive pathway required for the nutrient-sensitive interaction between raptor and mTOR. Mol Cell 11: 895–904.PubMedGoogle Scholar
  101. 101.
    Mordes DA, Glick GG, Zhao R, and Cortez D (2008) TopBP1 activates ATR through ATRIP and a PIKK regulatory domain. Genes Dev 22: 1478–1489.PubMedGoogle Scholar
  102. 102.
    de Klein A, Muijtjens M, van Os R, Verhoeven Y, Smit B, Carr AM, Lehmann AR, and Hoeijmakers JH (2000) Targeted disruption of the cell-cycle checkpoint gene ATR leads to early embryonic lethality in mice. Curr Biol 10: 479–482.PubMedGoogle Scholar
  103. 103.
    Brown EJ, and Baltimore D (2000) ATR disruption leads to chromosomal fragmentation and early embryonic lethality. Genes Dev 14: 397–402.PubMedGoogle Scholar
  104. 104.
    Paciotti V, Clerici M, Lucchini G, and Longhese MP (2000) The checkpoint protein Ddc2, functionally related to S. pombe Rad26, interacts with Mec1 and is regulated by Mec1-dependent phosphorylation in budding yeast. Genes Dev 14: 2046–2059.PubMedGoogle Scholar
  105. 105.
    Rouse J, and Jackson SP (2000) LCD1: an essential gene involved in checkpoint control and regulation of the MEC1 signalling pathway in Saccharomyces cerevisiae. Embo J 19: 5801–5812.PubMedGoogle Scholar
  106. 106.
    Wakayama T, Kondo T, Ando S, Matsumoto K, and Sugimoto K (2001) Pie1, a protein interacting with Mec1, controls cell growth and checkpoint responses in Saccharomyces cerevisiae. Mol Cell Biol 21: 755–764.PubMedGoogle Scholar
  107. 107.
    Edwards RJ, Bentley NJ, and Carr AM (1999) A Rad3-Rad26 complex responds to DNA damage independently of other checkpoint proteins. Nat Cell Biol 1: 393–398.PubMedGoogle Scholar
  108. 108.
    Cliby WA, Roberts CJ, Cimprich KA, Stringer CM, Lamb JR, Schreiber SL, and Friend SH (1998) Overexpression of a kinase-inactive ATR protein causes sensitivity to DNA-damaging agents and defects in cell cycle checkpoints. Embo J 17: 159–169.PubMedGoogle Scholar
  109. 109.
    Wright JA, Keegan KS, Herendeen DR, Bentley NJ, Carr AM, Hoekstra MF, and Concannon P (1998) Protein kinase mutants of human ATR increase sensitivity to UV and ionizing radiation and abrogate cell cycle checkpoint control. Proc Natl Acad Sci USA 95: 7445–7450.PubMedGoogle Scholar
  110. 110.
    Tibbetts RS, Brumbaugh KM, Williams JM, Sarkaria JN, Cliby WA, Shieh SY, Taya Y, Prives C, and Abraham RT (1999) A role for ATR in the DNA damage-induced phosphorylation of p53. Genes Dev 13: 152–157.PubMedGoogle Scholar
  111. 111.
    Guo Z, Kumagai A, Wang SX, and Dunphy WG (2000) Requirement for Atr in phosphorylation of Chk1 and cell cycle regulation in response to DNA replication blocks and UV-damaged DNA in Xenopus egg extracts. Genes Dev 14: 2745–2756.PubMedGoogle Scholar
  112. 112.
    Nghiem P, Park PK, Kim Y, Vaziri C, and Schreiber SL (2001) ATR inhibition selectively sensitizes G1 checkpoint-deficient cells to lethal premature chromatin condensation. Proc Natl Acad Sci USA 98: 9092–9097.PubMedGoogle Scholar
  113. 113.
    Liu Q, Guntuku S, Cui XS, Matsuoka S, Cortez D, Tamai K, Luo G, Carattini-Rivera S, DeMayo F, Bradley A, Donehower LA, and Elledge SJ (2000) Chk1 is an essential kinase that is regulated by Atr and required for the G(2)/M DNA damage checkpoint. Genes Dev 14: 1448–1459.PubMedGoogle Scholar
  114. 114.
    Cimprich KA, and Cortez D (2008) ATR: an essential regulator of genome integrity. Nat Rev Mol Cell Biol 9: 616–627.PubMedGoogle Scholar
  115. 115.
    Garvik B, Carson M, and Hartwell L (1995) Single-stranded DNA arising at telomeres in cdc13 mutants may constitute a specific signal for the RAD9 checkpoint. Mol Cell Biol 15: 6128–6138.PubMedGoogle Scholar
  116. 116.
    Costanzo V, and Gautier J (2003) Single-strand DNA gaps trigger an ATR- and Cdc7-dependent checkpoint. Cell Cycle 2: 17.PubMedGoogle Scholar
  117. 117.
    Costanzo V, Shechter D, Lupardus PJ, Cimprich KA, Gottesman M, and Gautier J (2003) An ATR- and Cdc7-dependent DNA damage checkpoint that inhibits initiation of DNA replication. Mol Cell 11: 203–213.PubMedGoogle Scholar
  118. 118.
    Zou L, and Elledge SJ (2003) Sensing DNA damage through ATRIP recognition of RPA-ssDNA complexes. Science 300: 1542–1548.PubMedGoogle Scholar
  119. 119.
    Zou L, Liu D, and Elledge SJ (2003) Replication protein A-mediated recruitment and activation of Rad17 complexes. Proc Natl Acad Sci USA 100: 13827–13832.PubMedGoogle Scholar
  120. 120.
    Stokes MP, Van Hatten R, Lindsay HD, and Michael WM (2002) DNA replication is required for the checkpoint response to damaged DNA in Xenopus egg extracts. J Cell Biol 158: 863–872.PubMedGoogle Scholar
  121. 121.
    Melo JA, Cohen J, and Toczyski DP (2001) Two checkpoint complexes are independently recruited to sites of DNA damage in vivo. Genes Dev 15: 2809–2821.PubMedGoogle Scholar
  122. 122.
    Kondo T, Wakayama T, Naiki T, Matsumoto K, and Sugimoto K (2001) Recruitment of Mec1 and Ddc1 checkpoint proteins to double-strand breaks through distinct mechanisms. Science 294: 867–870.PubMedGoogle Scholar
  123. 123.
    Majka J, and Burgers PM (2003) Yeast Rad17/Mec3/Ddc1: A sliding clamp for the DNA damage checkpoint. Proc Natl Acad Sci USA 100: 2249–2254.PubMedGoogle Scholar
  124. 124.
    Ellison V, and Stillman B (2003) Biochemical Characterization of DNA Damage Checkpoint Complexes: Clamp Loader and Clamp Complexes with Specificity for 5' Recessed DNA. PLoS Biol 1: E33.PubMedGoogle Scholar
  125. 125.
    Bonilla CY, Melo JA, and Toczyski DP (2008) Colocalization of sensors is sufficient to activate the DNA damage checkpoint in the absence of damage. Mol Cell 30: 267–276.PubMedGoogle Scholar
  126. 126.
    Toledo LI, Murga M, Gutierrez-Martinez P, Soria R, and Fernandez-Capetillo O (2008) ATR signaling can drive cells into senescence in the absence of DNA breaks. Genes Dev 22: 297–302.PubMedGoogle Scholar
  127. 127.
    Indiani C, and O'Donnell M (2006) The replication clamp-loading machine at work in the three domains of life. Nat Rev Mol Cell Biol 7: 751–761.PubMedGoogle Scholar
  128. 128.
    Majka J, Niedziela-Majka A, and Burgers PM (2006) The checkpoint clamp activates Mec1 kinase during initiation of the DNA damage checkpoint. Mol Cell 24: 891–901.PubMedGoogle Scholar
  129. 129.
    Furuya K, Poitelea M, Guo L, Caspari T, and Carr AM (2004) Chk1 activation requires Rad9 S/TQ-site phosphorylation to promote association with C-terminal BRCT domains of Rad4TOPBP1. Genes Dev 18: 1154–1164.PubMedGoogle Scholar
  130. 130.
    Delacroix S, Wagner JM, Kobayashi M, Yamamoto K, and Karnitz LM (2007) The Rad9-Hus1-Rad1 (9-1-1) clamp activates checkpoint signaling via TopBP1. Genes Dev 21: 1472–1477.PubMedGoogle Scholar
  131. 131.
    St Onge RP, Besley BD, Pelley JL, and Davey S (2003) A role for the phosphorylation of hRad9 in checkpoint signaling. J Biol Chem 278: 26620–26628.PubMedGoogle Scholar
  132. 132.
    Lee J, Kumagai A, and Dunphy WG (2007) The Rad9-Hus1-Rad1 checkpoint clamp regulates interaction of TopBP1 with ATR. J Biol Chem 282: 28036–28044.PubMedGoogle Scholar
  133. 133.
    Kumagai A, Lee J, Yoo HY, and Dunphy WG (2006) TopBP1 activates the ATR-ATRIP complex. Cell 124: 943–955.PubMedGoogle Scholar
  134. 134.
    Nakada D, Hirano Y, and Sugimoto K (2004) Requirement of the Mre11 complex and exonuclease 1 for activation of the Mec1 signaling pathway. Mol Cell Biol 24: 10016–10025.PubMedGoogle Scholar
  135. 135.
    Ferrell JE, Jr. (2002) Self-perpetuating states in signal transduction: positive feedback, double-negative feedback and bistability. Curr Opin Cell Biol 14: 140–148.PubMedGoogle Scholar
  136. 136.
    Gurley KE, and Kemp CJ (2001) Synthetic lethality between mutation in Atm and DNA-PK(cs) during murine embryogenesis. Curr Biol 11: 191–194.PubMedGoogle Scholar
  137. 137.
    Hickson I, Zhao Y, Richardson CJ, Green SJ, Martin NM, Orr AI, Reaper PM, Jackson SP, Curtin NJ, and Smith GC (2004) Identification and characterization of a novel and specific inhibitor of the ataxia-telangiectasia mutated kinase ATM. Cancer Res 64: 9152–9159.PubMedGoogle Scholar
  138. 138.
    Kolas NK, Chapman JR, Nakada S, Ylanko J, Chahwan R, Sweeney FD, Panier S, Mendez M, Wildenhain J, Thomson TM, Pelletier L, Jackson SP, and Durocher D (2007) Orchestration of the DNA-damage response by the RNF8 ubiquitin ligase. Science 318: 1637–1640.PubMedGoogle Scholar
  139. 139.
    Vaze MB, Pellicioli A, Lee SE, Ira G, Liberi G, Arbel-Eden A, Foiani M, and Haber JE (2002) Recovery from checkpoint-mediated arrest after repair of a double- strand break requires Srs2 helicase. Mol Cell 10: 373–385.PubMedGoogle Scholar
  140. 140.
    d'Adda di Fagagna F, Reaper PM, Clay-Farrace L, Fiegler H, Carr P, Von Zglinicki T, Saretzki G, Carter NP, and Jackson SP (2003) A DNA damage checkpoint response in telomere-initiated senescence. Nature 426: 194–198.PubMedGoogle Scholar
  141. 141.
    Bartkova J, Rezaei N, Liontos M, Karakaidos P, Kletsas D, Issaeva N, Vassiliou LV, Kolettas E, Niforou K, Zoumpourlis VC, Takaoka M, Nakagawa H, Tort F, Fugger K, Johansson F, Sehested M, Andersen CL, Dyrskjot L, Orntoft T, Lukas J, Kittas C, Helleday T, Halazonetis TD, Bartek J, and Gorgoulis VG (2006) Oncogene-induced senescence is part of the tumorigenesis barrier imposed by DNA damage checkpoints. Nature 444: 633–637.PubMedGoogle Scholar
  142. 142.
    Di Micco R, Fumagalli M, Cicalese A, Piccinin S, Gasparini P, Luise C, Schurra C, Garre M, Nuciforo PG, Bensimon A, Maestro R, Pelicci PG, and d'Adda di Fagagna F (2006) Oncogene-induced senescence is a DNA damage response triggered by DNA hyper-replication. Nature 444: 638–642.PubMedGoogle Scholar
  143. 143.
    Mallette FA, Gaumont-Leclerc MF, and Ferbeyre G (2007) The DNA damage signaling pathway is a critical mediator of oncogene-induced senescence. Genes Dev 21: 43–48.PubMedGoogle Scholar
  144. 144.
    Bartkova J, Horejsi Z, Koed K, Kramer A, Tort F, Zieger K, Guldberg P, Sehested M, Nesland JM, Lukas C, Orntoft T, Lukas J, and Bartek J (2005) DNA damage response as a candidate anti-cancer barrier in early human tumorigenesis. Nature 434: 864–870.PubMedGoogle Scholar
  145. 145.
    Gorgoulis VG, Vassiliou LV, Karakaidos P, Zacharatos P, Kotsinas A, Liloglou T, Venere M, Ditullio RA, Jr., Kastrinakis NG, Levy B, Kletsas D, Yoneta A, Herlyn M, Kittas C, and Halazonetis TD (2005) Activation of the DNA damage checkpoint and genomic instability in human precancerous lesions. Nature 434: 907–913.PubMedGoogle Scholar
  146. 146.
    Bartek J, Bartkova J, and Lukas J (2007) DNA damage signalling guards against activated oncogenes and tumour progression. Oncogene 26: 7773–7779.PubMedGoogle Scholar
  147. 147.
    Rogakou EP, Pilch DR, Orr AH, Ivanova VS, and Bonner WM (1998) DNA double-stranded breaks induce histone H2AX phosphorylation on serine 139. J Biol Chem 273: 5858–5868.PubMedGoogle Scholar
  148. 148.
    Ward IM, and Chen J (2001) Histone H2AX is phosphorylated in an ATR-dependent manner in response to replicational stress. J Biol Chem 276: 47759–47762.PubMedGoogle Scholar
  149. 149.
    Rogakou EP, Boon C, Redon C, and Bonner WM (1999) Megabase chromatin domains involved in DNA double-strand breaks in vivo. J Cell Biol 146: 905–916.PubMedGoogle Scholar
  150. 150.
    Olive PL (2004) Detection of DNA damage in individual cells by analysis of histone H2AX phosphorylation. Methods Cell Biol 75: 355–373.PubMedGoogle Scholar
  151. 151.
    Teicher BA (2008) Next generation topoisomerase I inhibitors: Rationale and biomarker strategies. Biochem Pharmacol 75: 1262–1271.PubMedGoogle Scholar
  152. 152.
    Ichijima Y, Sakasai R, Okita N, Asahina K, Mizutani S, and Teraoka H (2005) Phosphorylation of histone H2AX at M phase in human cells without DNA damage response. Biochem Biophys Res Commun 336: 807–812.PubMedGoogle Scholar
  153. 153.
    McManus KJ, and Hendzel MJ (2005) ATM-dependent DNA damage-independent mitotic phosphorylation of H2AX in normally growing mammalian cells. Mol Biol Cell 16: 5013–5025.PubMedGoogle Scholar
  154. 154.
    Ziv Y, Bielopolski D, Galanty Y, Lukas C, Taya Y, Schultz DC, Lukas J, Bekker-Jensen S, Bartek J, and Shiloh Y (2006) Chromatin relaxation in response to DNA double-strand breaks is modulated by a novel ATM- and KAP-1 dependent pathway. Nat Cell Biol 8: 870–876.PubMedGoogle Scholar
  155. 155.
    O'Connor MJ, Martin NM, and Smith GC (2007) Targeted cancer therapies based on the inhibition of DNA strand break repair. Oncogene 26: 7816–7824.PubMedGoogle Scholar
  156. 156.
    Ding J, Miao ZH, Meng LH, and Geng MY (2006) Emerging cancer therapeutic opportunities target DNA-repair systems. Trends Pharmacol Sci 27: 338–344.PubMedGoogle Scholar
  157. 157.
    Janetka JW, Ashwell S, Zabludoff S, and Lyne P (2007) Inhibitors of checkpoint kinases: from discovery to the clinic. Curr Opin Drug Discov Devel 10: 473–486.PubMedGoogle Scholar
  158. 158.
    Schaffner C, Idler I, Stilgenbauer S, Dohner H, and Lichter P (2000) Mantle cell lymphoma is characterized by inactivation of the ATM gene. Proc Natl Acad Sci USA 97: 2773–2778.PubMedGoogle Scholar
  159. 159.
    Greenman C, Stephens P, Smith R, Dalgliesh GL, Hunter C, Bignell G, Davies H, Teague J, Butler A, Stevens C, Edkins S, O'Meara S, Vastrik I, Schmidt EE, Avis T, Barthorpe S, Bhamra G, Buck G, Choudhury B, Clements J, Cole J, Dicks E, Forbes S, Gray K, Halliday K, Harrison R, Hills K, Hinton J, Jenkinson A, Jones D, Menzies A, Mironenko T, Perry J, Raine K, Richardson D, Shepherd R, Small A, Tofts C, Varian J, Webb T, West S, Widaa S, Yates A, Cahill DP, Louis DN, Goldstraw P, Nicholson AG, Brasseur F, Looijenga L, Weber BL, Chiew YE, DeFazio A, Greaves MF, Green AR, Campbell P, Birney E, Easton DF, Chenevix-Trench G, Tan MH, Khoo SK, Teh BT, Yuen ST, Leung SY, Wooster R, Futreal PA, and Stratton MR (2007) Patterns of somatic mutation in human cancer genomes. Nature 446: 153–158.PubMedGoogle Scholar
  160. 160.
    Dupre A, Boyer-Chatenet L, Sattler RM, Modi AP, Lee JH, Nicolette ML, Kopelovich L, Jasin M, Baer R, Paull TT, and Gautier J (2008) A forward chemical genetic screen reveals an inhibitor of the Mre11-Rad50-Nbs1 complex. Nat Chem Biol 4: 119–125.PubMedGoogle Scholar
  161. 161.
    Welch EM, Barton ER, Zhuo J, Tomizawa Y, Friesen WJ, Trifillis P, Paushkin S, Patel M, Trotta CR, Hwang S, Wilde RG, Karp G, Takasugi J, Chen G, Jones S, Ren H, Moon YC, Corson D, Turpoff AA, Campbell JA, Conn MM, Khan A, Almstead NG, Hedrick J, Mollin A, Risher N, Weetall M, Yeh S, Branstrom AA, Colacino JM, Babiak J, Ju WD, Hirawat S, Northcutt VJ, Miller LL, Spatrick P, He F, Kawana M, Feng H, Jacobson A, Peltz SW, and Sweeney HL (2007) PTC124 targets genetic disorders caused by nonsense mutations. Nature 447: 87–91.PubMedGoogle Scholar
  162. 162.
    Clerici M, Mantiero D, Lucchini G, and Longhese MP (2006) The Saccharomyces cerevisiae Sae2 protein negatively regulates DNA damage checkpoint signalling. EMBO Rep 7: 212–218.PubMedGoogle Scholar
  163. 163.
    Baldo V, Testoni V, Lucchini G, and Longhese MP (2008) Dominant TEL1-hy mutations compensate for Mec1 lack of functions in the DNA damage response. Mol Cell Biol 28: 358–375.PubMedGoogle Scholar
  164. 164.
    Morales M, Theunissen JW, Kim CF, Kitagawa R, Kastan MB, and Petrini JH (2005) The Rad50S allele promotes ATM-dependent DNA damage responses and suppresses ATM deficiency: implications for the Mre11 complex as a DNA damage sensor. Genes Dev 19: 3043–3054.PubMedGoogle Scholar
  165. 165.
    Alani E, Padmore R, and Kleckner N (1990) Analysis of wild-type and rad50 mutants of yeast suggests an intimate relationship between meiotic chromosome synapsis and recombination. Cell 61: 419–436.PubMedGoogle Scholar
  166. 166.
    Cao L, Alani E, and Kleckner N (1990) A pathway for generation and processing of double-strand breaks during meiotic recombination in S. cerevisiae. Cell 61: 1089–1101.PubMedGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2009

Authors and Affiliations

  1. 1.Samuel Lunenfeld Research InstituteCentre for Systems Biology, Mount Sinai HospitalTorontoCanada

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