Preferences help
enabled [disable] Abstract
Number of results
2007 | 54 | 1 | 63-69
Article title

Sirtuin inhibition increases the rate of non-homologous end-joining of DNA double strand breaks

Title variants
Languages of publication
Sirtuins (type III histone deacetylases) are an important member of a group of enzymes that modify chromatin conformation. We investigated the role of sirtuin inhibitor, GPI 19015, in double strand break (DSB) repair in CHO-K1 wt and xrs-6 mutant cells. The latter is defective in DNA-dependent protein kinase (DNA-PK)-mediated non-homologous end-joining (D-NHEJ). DSB were estimated by the neutral comet assay and histone γH2AX foci formation. We observed a weaker effect of GPI 19015 treatment on the repair kinetics in CHO wt cells than in xrs6. In the latter cells the increase in DNA repair rate was most pronounced in G1 phase and practically absent in S and G2 cell cycle phases. The decrease in the number of histone γH2AX foci was faster in xrs6 than in CHO-K1 cells. The altered repair rate did not affect survival of X-irradiated cells. Since in G1 xrs6 cells DNA-PK-dependent non-homologous end-joining, D-NHEJ, does not operate, these results indicate that inhibition of sirtuins modulates DNA-PK-independent (backup) non-homologous end-joining, B-NHEJ, to a greater extent than the other DSB repair system, D-NHEJ.
Physical description
  • Department of Radiobiology and Health Protection, Institute of Nuclear Chemistry and Technology, Warszawa, Poland
  • Department of Radiobiology and Health Protection, Institute of Nuclear Chemistry and Technology, Warszawa, Poland
  • Department of Radiobiology and Health Protection, Institute of Nuclear Chemistry and Technology, Warszawa, Poland
  • MGI Pharma, Inc., Baltimore, USA
  • MGI Pharma, Inc., Baltimore, USA
  • MGI Pharma, Inc., Baltimore, USA
  • Department of Radiobiology and Health Protection, Institute of Nuclear Chemistry and Technology, Warszawa, Poland
  • Ai X, Parthun MR (2004) The nuclear Hat1p/Hat2p complex: a molecular link between type B histone acetyltransferases and chromatin assembly. Mol Cell 14: 195-205.
  • Antonelli F, Belli M, Cuttone G, Dini V, Esposito G, Simone G, Sorrentino E, Tabocchini MA (2005) Induction and repair of DNA double-strand breaks in human cells: dephosphorylation of histone H2AX and its inhibition by calyculin A. Radiat Res 164: 514-517.
  • Audebert M, Salles B, Calsou P (2004) Involvement of poly(ADP-ribose) polymerase-1 and XRCC1/DNA ligase III in an alternative route for DNA double-strand breaks rejoining. J Biol Chem 279: 55117-55126.
  • Banath JP, Macphail SH, Olive PL (2004) Radiation sensitivity, H2AX phosphorylation, and kinetics of repair of DNA strand breaks in irradiated cervical cancer cell lines. Cancer Res 64: 7144-7149.
  • Buraczewska I, Bouzyk E, Kuduk-Jaworska J, Waszkiewicz K, Gasinska A, Szumiel I (2000) Differential anti-proliferative properties of novel hydroxydicarboxylatopla-tinum(II) complexes with high or low reactivity with thiols. Chem-Biol Interact 129: 297-315.
  • Camphausen K, Burgan W, Cerra M, Oswald KA, Trepel JB, Lee MJ, Tofilon PJ (2004a) Enhanced radiation-induced cell killing and prolongation of gammaH2AX foci expression by the histone deacetylase inhibitor MS-275. Cancer Res 64: 316-321.
  • Camphausen K, Scott T, Sproull M, Tofilon PJ (2004b) Enhancement of xenograft tumor radiosensitivity by the histone deacetylase inhibitor MS-275 and correlation with histone hyperacetylation. Clin Cancer Res 10: 6066-6071.
  • Camphausen K, Cerna D, Scott T, Sproull M, Burgan WE, Cerra MA, Fine H, Tofilon PJ (2005) Enhancement of in vitro and in vivo tumor cell radiosensitivity by valproic acid. Int J Cancer 114: 380-386.
  • Chakalova L, Russev G (1998) Comparison of repair activity in different genomic regions. Acta Biochim Polon 45: 173-81.
  • Cowell IG, Durkacz BW, Tilby MJ (2005) Sensitization of breast carcinoma cells to ionizing radiation by small molecule inhibitors of DNA-dependent protein kinase and ataxia telangiectasia mutated. Biochem Pharmacol 71: 13-20.
  • Goh M, Chen F, Paulsen MT, Yeager AM, Dyer ES, Ljungman M (2001) Phenylbutyrate attenuates the expression of Bcl-X(L), DNA-PK, caveolin-1, and VEGF in prostate cancer cells. Neoplasia 3: 331-338.
  • Kruszewski M, Szumiel I (2005) Sirtuins (histone deacetylases III) in the cellular response to DNA damage - facts and hypotheses. DNA Repair (Amst) 4: 1306-1313.
  • Kruszewski M, Wojewodzka M, Iwanenko T, Szumiel I, Okuyama A (1998) Differential inhibitory effect of OK-1035 on DNA repair in L5178Y murine lymphoma sublines with functional or defective repair of double strand breaks. Mutat Res 409: 31-36.
  • Linger J, Tyler JK (2005) The yeast histone chaperone chromatin assembly factor 1 protects against double-strand DNA-damaging agents. Genetics 171: 1513-1522.
  • Matsushita N, Takami Y, Kimura M, Tachiiri S, Ishiai M, Nakayama T, Takata M (2005) Role of NAD-dependent deacetylases SIRT1 and SIRT2 in radiation and cisplatin-induced cell death in vertebrate cells. Genes Cells 10: 321-332.
  • Morrison AJ, Shen X (2005) DNA repair in the context of chromatin. Cell Cycle 4: 568-571.
  • Munshi A, Kurland JF, Nishikawa T, Tanaka T, Hobbs ML, Tucker SL, Ismail S, Stevens C, Meyn RE (2005) Histone deacetylase inhibitors radiosensitize human melanoma cells by suppressing DNA repair activity. Clin Cancer Res 11: 4912-4922.
  • Perrault R, Wang H, Wang M, Rosidi B, Iliakis G (2004) Backup pathways of NHEJ are suppressed by DNA-PK. J Cell Biochem 92: 781-794.
  • Qin S, Parthun MR (2002) Histone H3 and the histone acetyltransferase Hat1p contribute to DNA double-strand break repair. Mol Cell Biol 22: 8353-8365.
  • Sak A, Stuschke M (1998) Repair of ionizing radiation induced DNA double-strand breaks (dsb) at the c-myc locus in comparison to the overall genome. Int J Radiat Biol 73: 35-43.
  • Seo H, Masuoka M, Murofushi H, Takeda S, Shibata T, Ohta K (2005) Rapid generation of specific antibodies by enhanced homologous recombination. Nat Biotechnol 23: 731-735.
  • Sun Y, Jiang X, Chen S, Fernandes N, 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.
  • Taneja N, Davis M, Choy JS, Beckett MA, Singh R, Kron SJ, Weichselbaum RR (2004) Histone H2AX phosphorylation as a predictor of radiosensitivity and target for radiotherapy. J Biol Chem 279: 2273-2280.
  • Tyler JK (2002) Chromatin assembly. Cooperation between histone chaperones and ATP-dependent nucleosome remodeling machines. Eur J Biochem 269: 2268-2274.
  • Wang H, Perrault AR, Takeda Y, Qin W, Wang H, Iliakis G (2003) Biochemical evidence for Ku-independent backup pathways of NHEJ. Nucleic Acids Res 31: 5377-5388.
  • Wang H, Rosidi B, Perrault R, Wang M, Zhang L, Windhofer F, Iliakis G (2005) DNA ligase III as a candidate component of backup pathways of nonhomologous end joining. Cancer Res 65: 4020-4030.
  • Wojewodzka M, Buraczewska I, Kruszewski M (2002) A modified neutral comet assay: elimination of lysis at high temperature and validation of the assay with anti-single-stranded DNA antibody. Mutat Res 518: 9-20.
  • Yaneva M, Li H, Marple T, Hasty P (2005) Non-homologous end joining, but not homologous recombination, enables survival for cells exposed to a histone deacetylase inhibitor. Nucleic Acids Res 33: 5320-5330.
  • Zhang J, Xu W (2005) Methods of NAD+-dependent deacetylase inhibitors. Patent Publication WO 2005053609.
Document Type
Publication order reference
YADDA identifier
JavaScript is turned off in your web browser. Turn it on to take full advantage of this site, then refresh the page.