Browse > Article
http://dx.doi.org/10.5487/TR.2008.24.3.175

Activation Mechanism of Protein Kinase B by DNA-dependent Protein Kinase Involved in the DNA Repair System  

Li, Yuwen (Department of Pharmacology, Cell Signaling Laboratory, Research Center for Transgenic Cloned Pigs, Daejeon Regional Cancer Center, Cancer Research Institute, Research Institute for Medical Sciences)
Piao, Longzhen (Department of Pharmacology, Cell Signaling Laboratory, Research Center for Transgenic Cloned Pigs, Daejeon Regional Cancer Center, Cancer Research Institute, Research Institute for Medical Sciences)
Yang, Keum-Jin (Department of Pharmacology, Cell Signaling Laboratory, Research Center for Transgenic Cloned Pigs, Daejeon Regional Cancer Center, Cancer Research Institute, Research Institute for Medical Sciences)
Shin, Sang-Hee (Department of Pharmacology, Cell Signaling Laboratory, Research Center for Transgenic Cloned Pigs, Daejeon Regional Cancer Center, Cancer Research Institute, Research Institute for Medical Sciences)
Shin, Eul-Soon (Department of Pharmacology, Cell Signaling Laboratory, Research Center for Transgenic Cloned Pigs, Daejeon Regional Cancer Center, Cancer Research Institute, Research Institute for Medical Sciences)
Park, Kyung-Ah (Department of Pharmacology, Cell Signaling Laboratory, Research Center for Transgenic Cloned Pigs, Daejeon Regional Cancer Center, Cancer Research Institute, Research Institute for Medical Sciences)
Byun, Hee-Sun (Department of Pharmacology, Cell Signaling Laboratory, Research Center for Transgenic Cloned Pigs, Daejeon Regional Cancer Center, Cancer Research Institute, Research Institute for Medical Sciences)
Won, Min-Ho (Department of Pharmacology, Cell Signaling Laboratory, Research Center for Transgenic Cloned Pigs, Daejeon Regional Cancer Center, Cancer Research Institute, Research Institute for Medical Sciences)
Choi, Byung-Lyul (Department of Pharmacology, Cell Signaling Laboratory, Research Center for Transgenic Cloned Pigs, Daejeon Regional Cancer Center, Cancer Research Institute, Research Institute for Medical Sciences)
Lee, Hyun-Ji (Department of Pharmacology, Cell Signaling Laboratory, Research Center for Transgenic Cloned Pigs, Daejeon Regional Cancer Center, Cancer Research Institute, Research Institute for Medical Sciences)
Kim, Young-Rae (Department of Pharmacology, Cell Signaling Laboratory, Research Center for Transgenic Cloned Pigs, Daejeon Regional Cancer Center, Cancer Research Institute, Research Institute for Medical Sciences)
Hong, Jang-Hee (Department of Pharmacology, Cell Signaling Laboratory, Research Center for Transgenic Cloned Pigs, Daejeon Regional Cancer Center, Cancer Research Institute, Research Institute for Medical Sciences)
Hur, Gang-Min (Department of Pharmacology, Cell Signaling Laboratory, Research Center for Transgenic Cloned Pigs, Daejeon Regional Cancer Center, Cancer Research Institute, Research Institute for Medical Sciences)
Kim, Jeong-Lan (Department of Psychiatry, College of Medicine, Chungnam National University)
Cho, Jae-Youl (School of Bioscience and Biotechnology, and Institute of Bioscience and Biotechnology, Kangwon National University)
Seok, Jeong-Ho (Department of Pharmacology, Cell Signaling Laboratory, Research Center for Transgenic Cloned Pigs, Daejeon Regional Cancer Center, Cancer Research Institute, Research Institute for Medical Sciences)
Park, Jong-Sun (Department of Pharmacology, Cell Signaling Laboratory, Research Center for Transgenic Cloned Pigs, Daejeon Regional Cancer Center, Cancer Research Institute, Research Institute for Medical Sciences)
Publication Information
Toxicological Research / v.24, no.3, 2008 , pp. 175-182 More about this Journal
Abstract
DNA-dependent protein kinase(DNA-PK) is involved in joining DNA double-strand breaks induced by ionizing radiation or V(D)J recombination and is activated by DNA ends and composed of a DNA binding subunit, Ku, and a catalytic subunit, DNA-PKcs. It has been suggested that DNA-PK might be $2^{nd}$ upstream kinase for protein kinase B(PKB). In this report, we showed that Ser473 phosphorylation in the hydrophobic-motif of PKB is blocked in DNA-PK knockout mouse embryonic fibroblast cells(MEFs) following insulin stimulation, while there is no effect on Ser473 phosphorylation in DNA-PK wild type MEF cells. The observation is further confirmed in human glioblastoma cells expressing a mutant form of DNA-PK(M059J) and a wild-type of DNA-PK(M059K), indicating that DNA-PK is indeed important for PKB activation. Furthermore, the treatment of cells with doxorubicin, DNA-damage inducing agent, leads to PKB phosphorylation on Ser473 in control MEF cells while there is no response in DNA-PK knockout MEF cells. Together, these results proposed that DNA-PK has a potential role in insulin signaling as well as DNA-repair signaling pathway.
Keywords
DNA-PK; DNA damage; Protein kinase B; Insulin signaling; Cell Signaling;
Citations & Related Records
연도 인용수 순위
  • Reference
1 Andjelkovic, M., Maira, S.M., Cron, P., Parker, P.J. and Hemmings, B.A. (1999). Domain swapping used to investigate the mechanism of protein kinase B regulation by 3- phosphoinositide-dependent protein kinase 1 and Ser473 kinase. Mol. Cell. Biol., 19, 5061-5072   DOI
2 Burma, S. and Chen, D.J. (2004). Role of DNA-PK in the cellular response to DNA double-strand breaks. DNA Repair (Amst), 3, 909-918   DOI   ScienceOn
3 Feng, J., Park, J., Cron, P., Hess, D. and Hemmings, B.A. (2004). Identification of a PKB/Akt hydrophobic motif ser- 473 kinase as DNA-dependent protein kinase. J. Biol. Chem., 279, 41189-41196   DOI   ScienceOn
4 Gottschalk, A.R., Doan, A., Nakamura, J.L., Stokoe, D. and Haas-Kogan, D.A. (2005). Inhibition of phosphatidylinositol- 3-kinase causes increased sensitivity to radiation through a PKB-dependent mechanism. Int. J. Radiat. Oncol. Biol. Phys., 63, 1221-1227   DOI   ScienceOn
5 Grana, T.M., Rusyn, E.V., Zhou, H., Sartor, C.I. and Cox, A.D. (2002). Ras mediates radioresistance through both phosphatidylinositol 3-kinase-dependent and Raf-dependent but mitogen-activated protein kinase/extracellular signalregulated kinase kinase-independent signaling pathways. Cancer. Res., 62, 4142-4150
6 Gupta, A.K., Cerniglia, G.J., Mick, R., Ahmed, M.S., Bakanauskas, V.J., Muschel, R.J. and McKenna, W.G. (2003). Radiation sensitization of human cancer cells in vivo by inhibiting the activity of PI3K using LY294002. Int. J. Radiat. Oncol. Biol. Phys., 56, 846-853   DOI   ScienceOn
7 Hanakahi, L.A., Bartlet-Jones, M., Chappell, C., Pappin, D. and West, S.C. (2000). Binding of inositol phosphate to DNA-PK and stimulation of double-strand break repair. Cell., 102, 721-729   DOI   ScienceOn
8 Hoeijmakers, J.H. (2001a). DNA repair mechanisms. Maturitas, 38, 17-22; discussion 22-13   DOI   ScienceOn
9 Ma, Y. and Lieber, M.R. (2002). Binding of inositol hexakisphosphate (IP6) to Ku but not to DNA-PKcs. J. Biol. Chem., 277, 10756-10759   DOI   ScienceOn
10 Stokoe, D., Stephens, L.R., Copeland, T., Gaffney, P.R., Reese, C.B., Painter, G.F., Holmes, A.B., McCormick, F. and Hawkins, P.T. (1997). Dual role of phosphatidylinositol-3,4,5-trisphosphate in the activation of protein kinase B. Science, 277, 567-570   DOI   ScienceOn
11 Yang, Z.Z., Tschopp, O., Baudry, A., Dummler, B., Hynx, D. and Hemmings, B.A. (2004). Physiological functions of protein kinase B/Akt. Biochem. Soc. Trans, 32, 350-354   DOI
12 Toker, A. and Newton, A.C. (2000). Akt/protein kinase B is regulated by autophosphorylation at the hypothetical PDK-2 site. J. Biol. Chem., 275, 8271-8274   DOI   ScienceOn
13 Allalunis-Turner, M.J., Barron, G.M., Day, R.S., 3rd, Dobler, K.D. and Mirzayans, R. (1993). Isolation of two cell lines from a human malignant glioma specimen differing in sensitivity to radiation and chemotherapeutic drugs. Radiat. Res., 134, 349-354   DOI
14 Dummler, B. and Hemmings, B.A. (2007). Physiological roles of PKB/Akt isoforms in development and disease. Biochem. Soc. Trans, 35, 231-235   DOI   ScienceOn
15 Hoeijmakers, J.H. (2001b). Genome maintenance mechanisms for preventing cancer. Nature, 411, 366-374   DOI   ScienceOn
16 Alessi, D.R., James, S.R., Downes, C.P., Holmes, A.B., Gaffney, P.R., Reese, C.B. and Cohen, P. (1997). Characterization of a 3-phosphoinositide-dependent protein kinase which phosphorylates and activates protein kinase Balpha. Curr. Biol., 7, 261-269   DOI   ScienceOn
17 Taccioli, G.E., Amatucci, A.G., Beamish, H.J., Gell, D., Xiang, X.H., Torres Arzayus, M.I., Priestley, A., Jackson, S.P., Marshak Rothstein, A., Jeggo, P.A. and Herrera, V.L. (1998). Targeted disruption of the catalytic subunit of the DNA-PK gene in mice confers severe combined immunodeficiency and radiosensitivity. Immunity, 9, 355-366   DOI   ScienceOn
18 Weterings, E. and van Gent, D.C. (2004). The mechanism of non-homologous end-joining: a synopsis of synapsis. DNA Repair (Amst), 3, 1425-1435   DOI   ScienceOn
19 Chan, D.W., Son, S.C., Block, W., Ye, R., Khanna, K.K., Wold, M.S., Douglas, P., Goodarzi, A.A., Pelley, J., Taya, Y., Lavin, M.F. and Lees-Miller, S.P. (2000). Purification and characterization of ATM from human placenta. A manganese-dependent, wortmannin-sensitive serine/threonine protein kinase. J. Biol. Chem., 275, 7803-7810   DOI   ScienceOn
20 O'Neill, T., Dwyer, A.J., Ziv, Y., Chan, D.W., Lees-Miller, S.P., Abraham, R.H., Lai, J.H., Hill, D., Shiloh, Y., Cantley, L.C. and Rathbun, G.A. (2000). Utilization of oriented peptide libraries to identify substrate motifs selected by ATM. J. Biol. Chem., 275, 22719-22727   DOI   ScienceOn
21 Byrum, J., Jordan, S., Safrany, S.T. and Rodgers, W. (2004). Visualization of inositol phosphate-dependent mobility of Ku: depletion of the DNA-PK cofactor InsP6 inhibits Ku mobility. Nucleic. Acids. Res., 32, 2776-2784   DOI   ScienceOn
22 Hanakahi, L.A. and West, S.C. (2002). Specific interaction of IP6 with human Ku70/80, the DNA-binding subunit of DNA-PK. Embo. J., 21, 2038-2044   DOI   ScienceOn
23 Sarbassov, D.D., Guertin, D.A., Ali, S.M. and Sabatini, D.M. (2005). Phosphorylation and regulation of Akt/PKB by the rictor-mTOR complex. Science, 307, 1098-1101   DOI   ScienceOn
24 Quevedo, C., Kaplan, D.R. and Derry, W.B. (2007). AKT-1 regulates DNA-damage-induced germline apoptosis in C. elegans. Curr. Biol., 17, 286-292   DOI   ScienceOn
25 van Gent, D.C., Hoeijmakers, J.H. and Kanaar, R. (2001). Chromosomal stability and the DNA double-stranded break connection. Nat. Rev. Genet., 2, 196-206   DOI   ScienceOn
26 Hresko, R.C. and Mueckler, M. (2005). mTOR.RICTOR is the Ser473 kinase for Akt/protein kinase B in 3T3-L1 adipocytes. J. Biol. Chem., 280, 40406-40416   DOI   ScienceOn
27 Alessi, D.R., Andjelkovic, M., Caudwell, B., Cron, P., Morrice, N., Cohen, P. and Hemmings, B.A. (1996). Mechanism of activation of protein kinase B by insulin and IGF-1. Embo. J., 15, 6541-6551
28 Andjelkovic, M., Alessi, D.R., Meier, R., Fernandez, A., Lamb, N.J., Frech, M., Cron, P., Cohen, P., Lucocq, J.M. and Hemmings, B.A. (1997). Role of translocation in the activation and function of protein kinase B. J. Biol. Chem., 272, 31515-31524   DOI   ScienceOn
29 Meek, K., Gupta, S., Ramsden, D.A. and Lees-Miller, S.P. (2004). The DNA-dependent protein kinase: the director at the end. Immunol. Rev., 200, 132-141   DOI   ScienceOn
30 Beamish, H.J., Jessberger, R., Riballo, E., Priestley, A., Blunt, T., Kysela, B. and Jeggo, P.A. (2000). The C-terminal conserved domain of DNA-PKcs, missing in the SCID mouse, is required for kinase activity. Nucleic. Acids. Res., 28, 1506-1513   DOI
31 Hartley, K.O., Gell, D., Smith, G.C., Zhang, H., Divecha, N., Connelly, M.A., Admon, A., Lees-Miller, S.P., Anderson, C.W. and Jackson, S.P. (1995). DNA-dependent protein kinase catalytic subunit: a relative of phosphatidylinositol 3-kinase and the ataxia telangiectasia gene product. Cell., 82, 849-856   DOI   ScienceOn
32 Gupta, A.K., Bakanauskas, V.J., Cerniglia, G.J., Cheng, Y., Bernhard, E.J., Muschel, R.J. and McKenna, W.G. (2001). The Ras radiation resistance pathway. Cancer. Res., 61, 4278-4282
33 Brazil, D.P., Yang, Z.-Z. and Hemmings, B.A. (2004). Advances in protein kinase B signalling: AKTion on multiple fronts. Trends in Biochemical Sciences, 29, 233-242   DOI   ScienceOn
34 Citterio, E., Vermeulen, W. and Hoeijmakers, J.H. (2000). Transcriptional healing. Cell., 101, 447-450   DOI   ScienceOn
35 Park, J., Hill, M.M., Hess, D., Brazil, D.P., Hofsteenge, J. and Hemmings, B.A. (2001). Identification of tyrosine phosphorylation sites on 3-phosphoinositide-dependent protein kinase-1 and their role in regulating kinase activity. J. Biol. Chem., 276, 37459-37471   DOI   ScienceOn
36 Weterings, E. and Chen, D.J. (2007). DNA-dependent pro tein kinase in nonhomologous end joining: a lock with multiple keys? J. Cell. Biol., 179, 183-186   DOI   ScienceOn
37 Li, X., Lu, Y., Liang, K., Liu, B. and Fan, Z. (2005). Differential responses to doxorubicin-induced phosphorylation and activation of Akt in human breast cancer cells. Breast. Cancer. Res., 7, R589-597   DOI   ScienceOn
38 Hill, M.M., Andjelkovic, M., Brazil, D.P., Ferrari, S., Fabbro, D. and Hemmings, B.A. (2001). Insulin-stimulated protein kinase B phosphorylation on Ser-473 is independent of its activity and occurs through a staurosporine-insensitive kinase. J. Biol. Chem., 276, 25643-25646   DOI   ScienceOn
39 Caporali, S., Levati, L., Starace, G., Ragone, G., Bonmassar, E., Alvino, E. and D'Atri, S. (2008). AKT is activated in an ATR-dependent manner in response to temozolomide and confers protection against drug-induced cell growth inhibition. Mol. Pharmacol., 74, 173-183   DOI   ScienceOn
40 Stephens, L., Anderson, K., Stokoe, D., Erdjument-Bromage, H., Painter, G.F., Holmes, A.B., Gaffney, P.R., Reese, C.B., McCormick, F., Tempst, P., Coadwell, J. and Hawkins, P.T. (1998). Protein kinase B kinases that mediate phosphatidylinositol 3,4,5-trisphosphate-dependent activation of protein kinase B. Science, 279, 710-714   DOI   ScienceOn
41 Li, B., Yuan, M., Kim, I.A., Chang, C.M., Bernhard, E.J. and Shu, H.K. (2004). Mutant epidermal growth factor receptor displays increased signaling through the phosphatidylinositol- 3 kinase/AKT pathway and promotes radioresistance in cells of astrocytic origin. Oncogene, 23, 4594-4602   DOI   ScienceOn
42 Bradford, M.M. (1976). A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem., 72, 248-254   DOI   ScienceOn
43 Viniegra, J.G., Martinez, N., Modirassari, P., Losa, J.H., Parada Cobo, C., Lobo, V.J., Luquero, C.I., Alvarez-Vallina, L., Ramon y Cajal, S., Rojas, J.M. and Sanchez-Prieto, R. (2005). Full activation of PKB/Akt in response to insulin or ionizing radiation is mediated through ATM. J. Biol. Chem., 280, 4029-4036   DOI   ScienceOn
44 Loeb, L.A. (1991). Mutator phenotype may be required for multistage carcinogenesis. Cancer. Res., 51, 3075-3079