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2015 | 62 | 2 | 173-176
Article title

Global quantification of heterochromatin-associated histone methylations in cell lines with differential sensitivity to ionizing radiation

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Histone modifications are involved in the DNA damage response (DDR). Here, by utilizing an ELISA immunoassay we assessed the methylation at H3K9 (H3K9me2 and H3K9me3) in two cell lines with differential sensitivity to radiation-induced apoptosis, HeLa (sensitive) and MCF-7 (resistant). We found that DNA damage induction by γ-irradiation leads to considerable accumulation (up to 5-fold) of H3K9me2 and H3K9me3, but not of H4K20me3 (control modification) in MCF-7 cells (p<0.05). Interestingly, a lower dose (2 Gy) was more effective than 5 Gy. In HeLa cells a smaller effect (approx. 1.5-1.8-fold) was evident only at 5 Gy. In conclusion, our findings reveal that DNA damage leads to specific accumulation of H3K9me2 and H3K9me3 in a cell-type specific manner.
Physical description
  • Ayoub N, Jeyasekharan AD, Bernal JA, Venkitaraman AR (2008) HP1-beta mobilization promotes chromatin changes that initiate the DNA damage response. Nature 453: 682-686.
  • Chen T, Zhang Y, Guo WH, Meng MB, Mo XM, Lu Y (2010) Effects of heterochromatin in colorectal cancer stem cells on radiosensitivity. Chin J Cancer 29: 270-276.
  • Dai B, Dahmani F, Cichocki JA, Swanson LC, Rasmussen TP (2011) Detection of post-translational modifications on native intact nucleosomes by ELISA. J Vis Exp 50: 2593.
  • Dai B, Giardina C, Rasmussen TP (2013) Quantitation of nucleosome acetylation and other histone posttranslational modifications using microscale NU-ELISA. Methods Mol Biol 981: 167-176.
  • Dickey JS, Redon CE, Nakamura AJ, Baird BJ, Sedelnikova OA, Bonner WM (2009) H2AX: functional roles and potential applications. Chromosoma 118: 683-692.
  • Djuzenova CS, Elsner I, Katzer A, Worschech E, Distel LV, Flentje M, Polat B (2013) Radiosensitivity in breast cancer assessed by the histone γ-H2AX and 53BP1 foci. Radiat Oncol 8: 98.
  • El-Awady RA, Dikomey E, Dahm-Daphi J (2003) Radiosensitivity of human tumour cells is correlated with the induction but not with the repair of DNA double-strand breaks. Br J Cancer 89: 593-601.
  • Essmann F, Engels IH, Totzke G, Schulze-Osthoff K, Jänicke RU (2004) Apoptosis resistance of MCF-7 breast carcinoma cells to ionizing radiation is independent of p53 and cell cycle control but caused by the lack of caspase-3 and a caffeine-inhibitable event. Cancer Res 64: 7065-7072.
  • Falk M, Lukasova E, Gabrielova B, Ondrej V, Kozubek S (2007) Chromatin dynamics during DSB repair. Biochim Biophys Acta 1773: 1534-1545.
  • Farifteh F, Salehi M, Bandehpour M, Nariman M, Ghafari Novin M, Hosseini T, Nematollahi S, Noroozian M, Keshavarzi S, Hosseini A (2014) Histone modification of embryonic stem cells produced by somatic cell nuclear transfer and fertilized blastocysts. Cell J Winter 15: 316-323.
  • Friedl AA, Mazurek B, Seiler DM (2012) Radiation-induced alterations in histone modification patterns and their potential impact on short-term radiation effects. Front Oncol 2: 117.
  • Iacovoni JS, Caron P, Lassadi I, Nicolas E, Massip L, Trouche D, Legube G (2010) High-resolution profiling of gammaH2AX around DNA double strand breaks in the mammalian genome. EMBO J 29: 1446-14457.
  • Jänicke RU, Engels IH, Dunkern T, Kaina B, Schulze-Osthoff K, Porter AG (2001) Ionizing radiation but not anticancer drugs causes cell cycle arrest and failure to activate the mitochondrial death pathway in MCF-7 breast carcinoma cells. Oncogene 20: 5043-5053.
  • Kumar R, Horikoshi N, Singh M, Gupta A, Misra HS, Albuquerque K, Hunt CR, Pandita TK (2013) Chromatin modifications and the DNA damage response to ionizing radiation. Front Oncol 2: 214.
  • Löbrich M, Shibata A, Beucher A, Fisher A, Ensminger M, Goodarzi AA, Barton O, Jeggo PA (2010) GammaH2AX foci analysis for monitoring DNA double-strand break repair: strengths, limitations and optimization. Cell Cycle 9: 662-669.
  • Luzhna L, Golubov A, Ilnytskyy S, Chekhun VF, Kovalchuk O (2013) Molecular mechanisms of radiation resistance in doxorubicin-resistant breast adenocarcinoma cells. Int J Oncol 42: 1692-1708.
  • Maroschik B, Gürtler A, Krämer A, Rößler U, Gomolka M, Hornhardt S, Mörtl S, Friedl AA (2014) Radiation-induced alterations of histone post-translational modification levels in lymphoblastoid cell lines. Radiat Oncol 9: 15.
  • Murr R, Loizou JI, Yang YG, Cuenin C, Li H, Wang ZQ, Herceg Z (2006) Histone acetylation by Trrap-Tip60 modulates loading of repair proteins and repair of DNA double-strand breaks. Nat Cell Biol 8: 91-99.
  • Nagaria P, Robert C, Rassool FV (2013) DNA double-strand break response in stem cells: mechanisms to maintain genomic integrity. Biochim Biophys Acta 1830: 2345-2353.
  • Özgür E, Mert U, Isin M, Okutan M, Dalay N, Gezer U (2013) Differential expression of long non-coding RNAs during genotoxic stress-induced apoptosis in HeLa and MCF-7 cells. Clin Exp Med 13: 119-126.
  • Peng JC, Karpen GH (2009) Heterochromatic genome stability requires regulators of histone H3 K9 methylation. PLoS Genet 5: E1000435.
  • Peters AH, O'Carroll D, Scherthan H, Mechtler K, Sauer S, Schöfer C, Weipoltshammer K, Pagani M, Lachner M, Kohlmaier A, Opravil S, Doyle M, Sibilia M, Jenuwein T (2001) Loss of the Suv39h histone methyltransferases impairs mammalian heterochromatin and genome stability. Cell 107: 323-337.
  • Rea S, Eisenhaber F, O'Carroll D, Strahl BD, Sun ZW, Schmid M, Opravil S, Mechtler K, Ponting CP, Allis CD, Jenuwein T (2000) Regulation of chromatin structure by site-specific histone H3 methyltransferases. Nature 406: 593-599.
  • Rothkamm K, Horn S, Scherthan H, Rössler U, De Amicis A, Barnard S, Kulka U, Lista F, Meineke V, Braselmann H, Beinke C, Abend M (2013) Laboratory intercomparison on the γ-H2AX foci assay. Radiat Res 180: 149-155.
  • Schotta G, Lachner M, Sarma K, Ebert A, Sengupta R, Reuter G, Reinberg D, Jenuwein T (2004) A silencing pathway to induce H3-K9 and H4-K20 trimethylation at constitutive heterochromatin. Genes Dev 18: 1251-1262.
  • Seiler DM, Rouquette J, Schmid VJ, Strickfaden H, Ottmann C, Drexler GA, Mazurek B, Greubel C, Hable V, Dollinger G, Cremer T, Friedl AA (2011) Double-strand break-induced transcriptional silencing is associated with loss of tri-methylation at H3K4. Chromosome Res 19: 883-899.
  • Solovjeva LV, Svetlova MP, Chagin VO, Tomilin NV (2007) Inhibition of transcription at radiation-induced nuclear foci of phosphorylated histone H2AX in mammalian cells. Chromosome Res 15: 787-797.
  • Stante M, Minopoli G, Passaro F, Raia M, Vecchio LD, Russo T (2009) Fe65 is required for Tip60-directed histone H4 acetylation at DNA strand breaks. Proc Natl Acad Sci USA 106: 5093-5098.
  • Sun Y, Jiang X, Xu Y, Ayrapetov MK, Moreau LA, Whetstine JR, Price BD (2009) Histone H3 methylation links DNA damage detection to activation of the tumour suppressor Tip60. Nat Cell Biol 11: 1376-1382.
  • Tjeertes JV, Miller KM, Jackson SP (2009) Screen for DNA-damage-responsive histone modifications identifies H3K9Ac and H3K56Ac in human cells. EMBO J 28: 1878-1889.
  • Turner BM (2012) The adjustable nucleosome: an epigenetic signaling module. Trends Genet 28: 436-444.
  • Young LC, McDonald DW, Hendzel MJ (2013) Kdm4b histone demethylase is a DNA damage response protein and confers a survival advantage following γ-irradiation. J Biol Chem 288: 21376-21388.
  • Xu Y, Xu C, Price BD (2012) Mechanistic links between ATM and histone methylation codes during DNA repair. Prog Mol Biol Transl Sc 110: 263-288.
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