Abstract
DNA-dependent protein kinase (DNA-PK) is required for the repair of double strand DNA breaks by nonhomologous DNA end joining. The catalytic subunit of DNA-PK, PRKDC, may also be involved in repair-related or separate cell signaling pathways. To learn more about the cellular function of DNA-PK under normal physiological conditions, we identified genes that are differentially expressed between an immortalized wild-type mouse fibroblast cell line and its DNA-PK-deficient counterpart (Prkdc−/−). The proto-oncogene Mdm2 and the farnesoid X receptor gene Nr1h4 were overexpressed in the DNA-PK-deficient cell line. We show that in the DNA-PK-deficient cell line the genes for both Mdm2 and Nr1h4 are amplified to a degree that could account for most, if not all, of their increased expression. Other genes were strongly downregulated in the DNA-PK-deficient cell line, but this opposite expression pattern was not due to gene amplification in the wild-type cells. None of these genes was differentially expressed in DNA-PK-containing and DNA-PK-deficient primary mouse embryo fibroblasts. Our results suggest a model in which DNA-PK indirectly affects the cellular gene expression profile through its caretaker role and by preventing gene amplification.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Kirchgessner C.U., Patil C.K., Evans J.W., Cuomo C.A., Fried L.M., Carter T., Oettinger M.A. and Brown J.M. 1995. Science 267: 1178–1183.
Lees-Miller S.P., Godbout R., Chan D.W., Weinfeld M., Day R.S. 3rd, Barron G.M. and Allalunis-Turner J. 1995. Science 267: 1183–1185.
Kurimasa A., Kumano S., Boubnov N.V., Story M.D., Tung C.S., Peterson S.R. and Chen D.J. 1999. Mol. Cell. Biol. 19: 3877–3884.
Kharbanda S., Pandey P., Jin S., Inoue S., Bharti A., Yuan Z.-M., Weichselbaum R., Weaver D. and Kufe D. 1997. Nature 386: 732–735.
Woo R.A., McLure K.G., Lees-Miller S.P., Rancourt D.E. and Lee P.W.K. (1998) Nature 394: 700–704.
Galloway A.M. and Allalunis-Turner J. 2000. Radiat. Res. 154: 609–615.
Bryntesson F., Regan J.C., Jeggo P.A., Taccioli G.E. and Hubank M. 2001. Radiat. Res. 156: 167–176.
Ai R., Sandoval A. and Labhart P. (2003) Gene Expr. 11: 35–45.
Kurimasa A., Ouyang H., Dong L., Wang S., Li X., Cordon-Cardo C., Chen D.J. and Li, G.C. 1999. Proc. Natl. Acad. Sci. USA 96: 1403–1408.
Araki R., Fukumura R., Fujimori A., Taya Y., Shiloh Y., Kurimasa A., Burma S., Li G.C., Chen D.J., Sato K., Hoki Y., Tatsumi K. and Abe M. 1999. Cancer Res. 59: 3543–3546.
Walker A.I., Hunt T., Jackson R.J. and Anderson C.W. 1985. EMBO J. 4: 139–145.
Finnie N.J., Gottlieb T.M., Blunt T., Jeggo P. and Jackson S.P. 1995. Proc. Natl. Acad. Sci. USA 92: 320–324.
Welsh J., Chada K., Dalal S.S., Cheng R., Ralph D. and McClelland M. 1992. Nucleic. Acids. Res. 20: 4965–4970.
Seol W., Choi H.S. and Moore D.D. 1995. Mol. Endocrinol. 9: 72–85.
Olson D.C., Marechal V., Momand J., Chen J., Romocki C. and Levine A.J. 1993. Oncogene 8: 2353–2360.
Koike M. 2002. J. Radiat. Res. (Tokyo) 43: 223–236.
Fakharzadeh S.S., Trusko S.P. and George D.L. 1991. EMBO J. 10: 1565–1569.
Tlsty T.D. 1990. Proc. Natl. Acad. Sci. USA 87: 3132–3136.
Wright J.A., Smith H.S., Watt F.M., Hancok M.C., Hudson D.L. and Stark G.R. 1990. Proc. Natl. Acad. Sci. USA 87: 1791–1795.
Windle B., Draper B.W., Yin Y.X., O'Gorman S. and Wahl G.M. 1991. Genes. Dev. 5: 160–174.
Ma C., Martin S., Trask B. and Hamlin J.L. 1993. Genes. Dev. 7: 605–620.
Mondello C., Rebuzzini P., Dolzan M., Edmonson S., Taccioli G.E. and Giulotto E. 2001. Cancer Res. 61: 4520–4525.
Paulson T.G., Almasan A., Bordy L.L. and Wahl G.M. 1998. Mol. Cell. Biol. 18: 3089–3100.
Karanjawala Z.E., Grawunder U., Hsieh C.L. and Lieber M.R. 1999. Curr. Biol. 9: 1501–1504.
Difilippantonio M.J., Zhu J., Chen H.T., Meffre E., Nussenzweig M.C., Max E.E., Ried T. and Nussenzweig A. 2000. Nature 404: 510–514.
Zhu C., Mills K.D., Ferguson D.O., Lee C., Manis J., Fleming J., Gao Y., Morton C.C. and Alt F.W. 2002. Cell 109: 811–821.
Gladdy R.A., Taylor M.D., Williams C.J., Grandal I., Karaskova J., Squire J.A., Rutka J.T., Guidos C.J., and Danska J.S. 2003. Cancer Cell 3: 37–50.
Sharpless N.E., Ferguson D.O., O'Hagan R.C., Castrillon D.H., Lee C., Farazi P.A., Alson S., Fleming J., Morton C.C., Frank K., Chin L., Alt F.W. and DePinho R.A. 2001. Mol. Cell 8: 1187–1196.
Haines D.S. 1997. Leuk. Lymphoma 26: 227–238
Laffitte B.A., Kast H.R., Nguyen C.M., Zavacki A.M., Moore D.D. and Edwards P.A. 2000. J. Biol. Chem. 275: 10638–10647.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Ai, R., Sandoval, A., Chen, D.J. et al. Amplification and overexpression of oncogene Mdm2 and orphan receptor gene Nr1h4 in immortal PRKDC knockout cells. Mol Biol Rep 31, 91–96 (2004). https://doi.org/10.1023/B:MOLE.0000031358.71141.78
Issue Date:
DOI: https://doi.org/10.1023/B:MOLE.0000031358.71141.78