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Department of Radiation Medicine, Division of Radiation Research, Georgetown University Medical Center, Washington, DC 20007 [A. D. K., M. O. J.], and Medicine Branch, National Cancer Institute, NIH, Bethesda, Maryland 20892 [K-N. C.]
A radiation-inducible immediate-early gene, IEX-1, was identified and characterized in human squamous carcinoma cells. Sequence analysis revealed 156-amino acid nucleotides, encoding a protein of Mr 20,000. The protein is glycosylated (Mr 
27,000) in the presence of microsomal membranes. Northern analysis reveals a 1.2-kb transcript. Treatment with cycloheximide was associated with superinduction of this transcript, suggesting that it is an immediate-early gene. The abundance of IEX-1 mRNA increased rapidly after exposure of the cells to ionizing radiation (2–10 Gy), reaching a maximum by 15 min and returning subsequently to basal levels by 4 h. Expression of IEX-1 was also induced significantly by the protein kinase C (PKC) activator 12-O-tetradecanoylphorbol-13-acetate (TPA), the protein phosphatase inhibitor okadaic acid, and tumor necrosis factor-
, whereas treatment of cells with UV light and H202 had little effect on IEX-1 expression. Cells depleted of PKC by prolonged incubation with TPA showed no attenuated IEX-1 response to tumor necrosis factor-. This is the first report of IEX-1, a radiation-inducible glycosylated human protein, whose expression can be mediated through multisignal transduction pathways.
1 To whom requests for reprints should be addressed, at Department of Radiation Medicine, Division of Radiation Research, The Research Building, E211A, 3970 Reservoir Road NW, Washington, DC 20007. Phone: (202) 687-8352; Fax: (202) 687-2221.
Received 12/19/96. Accepted 2/13/96.
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