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Originally published In Press as doi:10.1074/jbc.M006190200 on November 15, 2000

J. Biol. Chem., Vol. 276, Issue 12, 8884-8891, March 23, 2001
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Epidermal Growth Factor Sensitizes Cells to Ionizing Radiation by Down-regulating Protein Mutated in Ataxia-Telangiectasia*

Nuri GuevenDagger §, Katherine E. Keating§, Philip Chen, Toshiyuki Fukao||, Kum Kum Khanna, Dianne Watters**, Peter H. RodemannDagger , and Martin F. LavinDagger Dagger

From the Dagger  Section for Radiobiology and Molecular Environmental Research, Röntgenweg 11, 72076 Tübingen, Germany, the  Queensland Cancer Fund Research Laboratory, The Queensland Institute of Medical Research, P. O. Royal Brisbane Hospital, Brisbane, Queensland 4029, Australia, the || Gifu University School of Medicine, 40 Tsukasa-Machi, Gifu 500-8076, Japan, and the ** Department of Surgery, University of Queensland, P. O. Royal Brisbane Hospital, Brisbane, Queensland 4029, Australia

Epidermal growth factor (EGF) has been reported to either sensitize or protect cells against ionizing radiation. We report here that EGF increases radiosensitivity in both human fibroblasts and lymphoblasts and down-regulates both ATM (mutated in ataxia-telangiectasia (A-T)) and the catalytic subunit of DNA-dependent protein kinase (DNA-PKcs). No further radiosensitization was observed in A-T cells after pretreatment with EGF. The down-regulation of ATM occurs at the transcriptional level. Concomitant with the down-regulation of ATM, the DNA binding activity of the transcription factor Sp1 decreased. A causal relationship was established between these observations by demonstrating that up-regulation of Sp1 DNA binding activity by granulocyte/macrophage colony-stimulating factor rapidly reversed the EGF-induced decrease in ATM protein and restored radiosensitivity to normal levels. Failure to radiosensitize EGF-treated cells to the same extent as observed for A-T cells can be explained by induction of ATM protein and kinase activity with time post-irradiation. Although ionizing radiation damage to DNA rapidly activates ATM kinase and cell cycle checkpoints, we have provided evidence for the first time that alteration in the amount of ATM protein occurs in response to both EGF and radiation exposure. Taken together these data support complex control of ATM function that has important repercussions for targeting ATM to improve radiotherapeutic benefit.

* This work was supported in part by the Australian National Health and Medical Research Council, the Deutsche Forschungsgemeinschaft Grant 488/1-1, and the A-T Children's foundation.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

§ Both authors contributed equally to this work.

Dagger Dagger To whom correspondence should be addressed: the Queensland Cancer Fund Research Laboratory, Queensland Institute of Medical Research, P. O. Royal Brisbane Hospital, Brisbane, Queensland 4029, Australia. Tel.: 61-7-33620341; Fax: 61-7-33620106; E-mail: martinL@qimr.edu.au.

Copyright © 2001 by The American Society for Biochemistry and Molecular Biology, Inc.
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