Cellular Responses to Anthracyclines Identify Ku70, a DNA Repair Factor that Changes Compartment and Remains Stable in Leukemic Cells
Abstract
Anthracyclines such as doxorubicin and daunorubicin are anticancer drugs that act by damaging the DNA and used for treating a variety of cancers including adult acute myeloid leukemia. To date, nearly 50 % of acute myeloid leukemia patients show resistance to anthracyclines although the cause is not known. We first investigate if there is a relationship between the expression level of 23 DNA repair genes in three leukemic cell lines (KG-1, HL-60 and Mono-Mac1) and cellular responses to anthracyclines. We observed that the DNA repair genes were all downregulated in these cell lines following exposure to doxorubicin. Further analysis revealed that the general downregulation of the genes was linked to a substantial decrease in the recovery of total RNA raising the possibility that assessment of total RNA, and not specific gene or set of genes, can be used as a simple indicator of cellular responses to anthracyclines. Furthermore, examination of total protein extracts derived from these cell lines revealed for the first time that Ku70 is a key protein that remained stable, while the majority of proteins were loss, upon anthracycline treatment. Importantly, Ku70 redistributes from the cytoplasm to the nucleoli in a time-dependent manner in response to anthracycline exposure. We propose that Ku70 redistribution might play a vital role in predicting cellular response to anthracycline and promoting cell death.
Keywords
Full Text:
PDFReferences
Dohner H, Estey EH, Amadori S, Appelbaum FR, Buchner T, Burnett AK, Dombret H, Fenaux P, Grimwade D, Larson RA, Lo-Coco F, Naoe T, Niederwieser D, Ossenkoppele GJ, Sanz MA, Sierra J, Tallman MS, Lowenberg B, Bloomfield CD. Diagnosis and management of acute myeloid leukemia in adults: Recommendations from an international expert panel, on behalf of the european leukemianet. Blood. 2010, 115:453-474
Fornari FA, Randolph JK, Yalowich JC, Ritke MK, Gewirtz DA. Interference by doxorubicin with DNA unwinding in mcf-7 breast tumor cells. Mol Pharmacol. 1994, 45:649-656
Momparler RL, Karon M, Siegel SE, Avila F. Effect of adriamycin on DNA, rna, and protein synthesis in cell-free systems and intact cells. Cancer Res. 1976, 36:2891-2895
Childs AC, Phaneuf SL, Dirks AJ, Phillips T, Leeuwenburgh C. Doxorubicin treatment in vivo causes cytochrome c release and cardiomyocyte apoptosis, as well as increased mitochondrial efficiency, superoxide dismutase activity, and bcl-2:Bax ratio. Cancer Res. 2002, 62:4592-4598
Forrest RA, Swift LP, Rephaeli A, Nudelman A, Kimura KI, Phillips DR, Cutts SM. Activation of DNA damage response pathways as a consequence of anthracycline-DNA adduct formation. Biochem Pharmacol. 2012,
Spencer DM, Bilardi RA, Koch TH, Post GC, Nafie JW, Kimura K, Cutts SM, Phillips DR. DNA repair in response to anthracycline-DNA adducts: A role for both homologous recombination and nucleotide excision repair. Mutat Res. 2008, 638:110-121
Moraes MC, de Andrade AQ, Carvalho H, Guecheva T, Agnoletto MH, Henriques JA, Sarasin A, Stary A, Saffi J, Menck CF. Both xpa and DNA polymerase eta are necessary for the repair of doxorubicin-induced DNA lesions. Cancer Lett. 2012, 314:108-118
Saffi J, Agnoletto MH, Guecheva TN, Batista LF, Carvalho H, Henriques JA, Stary A, Menck CF, Sarasin A. Effect of the anti-neoplastic drug doxorubicin on xpd-mutated DNA repair-deficient human cells. DNA Repair (Amst). 2010, 9:40-47
Liddiard K, Hills R, Burnett AK, Darley RL, Tonks A. Ogg1 is a novel prognostic indicator in acute myeloid leukaemia. Oncogene. 2010, 29:2005-2012
Zaremba T, Thomas H, Cole M, Plummer ER, Curtin NJ. Doxorubicin-induced suppression of poly(adp-ribose) polymerase-1 (parp-1) activity and expression and its implication for parp inhibitors in clinical trials. Cancer Chemother Pharmacol. 2010, 66:807-812
Noren Hooten N, Kompaniez K, Barnes J, Lohani A, Evans MK. Poly(adp-ribose) polymerase 1 (parp-1) binds to 8-oxoguanine-DNA glycosylase (ogg1). J Biol Chem. 2011, 286:44679-44690
Poschmann J, Drouin S, Jacques PE, El Fadili K, Newmarch M, Robert F, Ramotar D. The peptidyl prolyl isomerase rrd1 regulates the elongation of rna polymerase ii during transcriptional stresses. PLoS One. 2011, 6:e23159
Andrabi SA, Kim NS, Yu SW, Wang H, Koh DW, Sasaki M, Klaus JA, Otsuka T, Zhang Z, Koehler RC, Hurn PD, Poirier GG, Dawson VL, Dawson TM. Poly(adp-ribose) (par) polymer is a death signal. Proc Natl Acad Sci U S A. 2006, 103:18308-18313
Woodhouse BC, Dianov GL. Poly adp-ribose polymerase-1: An international molecule of mystery. DNA Repair (Amst). 2008, 7:1077-1086
Roberts SA, Strande N, Burkhalter MD, Strom C, Havener JM, Hasty P, Ramsden DA. Ku is a 5'-drp/ap lyase that excises nucleotide damage near broken ends. Nature. 2010, 464:1214-1217
Sawada M, Sun W, Hayes P, Leskov K, Boothman DA, Matsuyama S. Ku70 suppresses the apoptotic translocation of bax to mitochondria. Nat Cell Biol. 2003, 5:320-329
Muller C, Paupert J, Monferran S, Salles B. The double life of the ku protein: Facing the DNA breaks and the extracellular environment. Cell Cycle. 2005, 4:438-441
Featherstone C, Jackson SP. Ku, a DNA repair protein with multiple cellular functions? Mutat Res. 1999, 434:3-15
Essers J, Theil AF, Baldeyron C, van Cappellen WA, Houtsmuller AB, Kanaar R, Vermeulen W. Nuclear dynamics of pcna in DNA replication and repair. Mol Cell Biol. 2005, 25:9350-9359
Shivji KK, Kenny MK, Wood RD. Proliferating cell nuclear antigen is required for DNA excision repair. Cell. 1992, 69:367-374
Clark AB, Valle F, Drotschmann K, Gary RK, Kunkel TA. Functional interaction of proliferating cell nuclear antigen with msh2-msh6 and msh2-msh3 complexes. J Biol Chem. 2000, 275:36498-36501
Parker A, Gu Y, Mahoney W, Lee SH, Singh KK, Lu AL. Human homolog of the muty repair protein (hmyh) physically interacts with proteins involved in long patch DNA base excision repair. J Biol Chem. 2001, 276:5547-5555
Henneke G, Koundrioukoff S, Hubscher U. Phosphorylation of human fen1 by cyclin-dependent kinase modulates its role in replication fork regulation. Oncogene. 2003, 22:4301-4313
Gary R, Ludwig DL, Cornelius HL, MacInnes MA, Park MS. The DNA repair endonuclease xpg binds to proliferating cell nuclear antigen (pcna) and shares sequence elements with the pcna-binding regions of fen-1 and cyclin-dependent kinase inhibitor p21. J Biol Chem. 1997, 272:24522-24529
Fan J, Otterlei M, Wong HK, Tomkinson AE, Wilson DM, 3rd. Xrcc1 co-localizes and physically interacts with pcna. Nucleic Acids Res. 2004, 32:2193-2201
Matheos D, Ruiz MT, Price GB, Zannis-Hadjopoulos M. Ku antigen, an origin-specific binding protein that associates with replication proteins, is required for mammalian DNA replication. Biochim Biophys Acta. 2002, 1578:59-72
Pigram WJ, Fuller W, Hamilton LD. Stereochemistry of intercalation: Interaction of daunomycin with DNA. Nat New Biol. 1972, 235:17-19
Koike M, Miyasaka T, Mimori T, Shiomi T. Subcellular localization and protein-protein interaction regions of ku proteins. Biochem Biophys Res Commun. 1998, 252:679-685
Koike M, Shiomi T, Koike A. Ku70 can translocate to the nucleus independent of ku80 translocation and DNA-pk autophosphorylation. Biochem Biophys Res Commun. 2000, 276:1105-1111
Koike M, Awaji T, Kataoka M, Tsujimoto G, Kartasova T, Koike A, Shiomi T. Differential subcellular localization of DNA-dependent protein kinase components ku and DNA-pkcs during mitosis. J Cell Sci. 1999, 112 ( Pt 22):4031-4039
Mimori T, Hardin JA, Steitz JA. Characterization of the DNA-binding protein antigen ku recognized by autoantibodies from patients with rheumatic disorders. J Biol Chem. 1986, 261:2274-2278
Bakalkin G, Yakovleva T, Hurd YL, Nussenzweig A, Li GC, Terenius L. Autoantigen ku in the brain. Developmentally regulated expression and subcellular localization. Neuroreport. 1998, 9:2147-2151
Yaneva M, Jhiang S. Expression of the ku protein during cell proliferation. Biochim Biophys Acta. 1991, 1090:181-187
Prabhakar BS, Allaway GP, Srinivasappa J, Notkins AL. Cell surface expression of the 70-kd component of ku, a DNA-binding nuclear autoantigen. J Clin Invest. 1990, 86:1301-1305
Fewell JW, Kuff EL. Intracellular redistribution of ku immunoreactivity in response to cell-cell contact and growth modulating components in the medium. J Cell Sci. 1996, 109 ( Pt 7):1937-1946
Grawunder U, Finnie N, Jackson SP, Riwar B, Jessberger R. Expression of DNA-dependent protein kinase holoenzyme upon induction of lymphocyte differentiation and v(d)j recombination. Eur J Biochem. 1996, 241:931-940
Pucci S, Mazzarelli P, Sesti F, Boothman DA, Spagnoli LG. Interleukin-6 affects cell death escaping mechanisms acting on bax-ku70-clusterin interactions in human colon cancer progression. Cell Cycle. 2009, 8:473-481
Pucci S, Bonanno E, Pichiorri F, Angeloni C, Spagnoli LG. Modulation of different clusterin isoforms in human colon tumorigenesis. Oncogene. 2004, 23:2298-2304
Pucci S, Mazzarelli P, Nucci C, Ricci F, Spagnoli LG. Clu "in and out": Looking for a link. Adv Cancer Res. 2009, 105:93-113
Mazzarelli P, Pucci S, Spagnoli LG. Clu and colon cancer. The dual face of clu: From normal to malignant phenotype. Adv Cancer Res. 2009, 105:45-61
Moore HM, Bai B, Boisvert FM, Latonen L, Rantanen V, Simpson JC, Pepperkok R, Lamond AI, Laiho M. Quantitative proteomics and dynamic imaging of the nucleolus reveal distinct responses to uv and ionizing radiation. Mol Cell Proteomics. 2011, 10:M111 009241
Refbacks
- There are currently no refbacks.
