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J. Biol. Chem., Vol. 279, Issue 3, 1615-1620, January 16, 2004
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B Kinases Determines the Significance of Phosphatidylinositol 3-Kinase/Akt Signaling to NF-B Activation*
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From the
Department of Microbiology and Immunology and Walther Oncology Center, Indiana University School of Medicine, Indianapolis, Indiana 46202 and Walther Cancer Institute, Indianapolis, Indiana 46208 and the ¶Laboratory of Genetics, Salk Institute, La Jolla, California 92037
Phosphatidylinositol (PI) 3-kinase/Akt signaling activates NF-
B through pleiotropic, cell type-specific mechanisms. This study investigated the significance of PI 3-kinase/Akt signaling to tumor necrosis factor (TNF)-induced NF-B activation in transformed, immortalized, and primary cells. Pharmacological inhibition of PI 3-kinase blocked TNF-induced NF-B DNA binding in the 293 line of embryonic kidney cells, partially affected binding in MCF-7 breast cancer cells, HeLa and ME-180 cervical carcinoma cells, and NIH 3T3 cells but was without significant effect in H1299 and human umbilical vein endothelial cells, cell types in which TNF activated Akt. NF-B is retained in the cytoplasm by inhibitory proteins, IBs, which are phosphorylated and targeted for degradation by IB kinases (IKK
and IKK
). Expression and the ratios of IKK and IKK, which homo- and heterodimerize, varied among cell types. Cells with a high proportion of IKK (the IKK kinase activated by Akt) to IKK were most sensitive to PI 3-kinase inhibitors. Consequently, transient expression of IKK diminished the capacity of the inhibitors to block NF-B DNA binding in 293 cells. Also, inhibitors of PI 3-kinase blocked NF-B DNA binding in Ikk–/– but not Ikk–/– or wild-type cells in which the ratio of IKK to IKK is low. Thus, noncoordinate expression of IB kinases plays a role in determining the cell type-specific role of Akt in NF-B activation.
Received for publication, June 30, 2003 , and in revised form, October 27, 2003.
* This work was supported by National Institutes of Health Grants CA67891 and 73023 (to D. B. D.). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
Supported by the Indiana University Cancer Center and the Indiana University Diabetes Training Program.
|| To whom correspondence should be addressed: Walther Oncology Center, Indiana University School of Medicine, 950 West Walnut St., Indianapolis, IN 46202. E-mail: ddonner{at}iupui.edu.
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