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Originally published In Press as doi:10.1074/jbc.C000135200 on May 17, 2000

J. Biol. Chem., Vol. 275, Issue 27, 20255-20259, July 7, 2000
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Overexpression of a Nuclear Protein, TIEG, Mimics Transforming Growth Factor-beta Action in Human Osteoblast Cells*

Theresa E. HefferanDagger §, Gregory G. ReinholzDagger §, David J. RickardDagger , Steven A. JohnsenDagger , Katrina M. Waters, M. SubramaniamDagger , and Thomas C. SpelsbergDagger ||

From the Dagger  Department of Biochemistry and Molecular Biology, Mayo Clinic and Mayo Foundation, Rochester, Minnesota 55905 and the  Division of Endocrine, Reproductive, and Developmental Toxicology, Chemical Industry Institute of Toxicology, Research Triangle Park, North Carolina 27709

Although transforming growth factor-beta (TGF-beta ) is a growth factor with many known regulatory activities in many different cell types, its intracellular signaling pathway is still not fully understood. A TGF-beta -inducible early gene (TIEG) was discovered and shown by this laboratory to be a 3-zinc finger transcription factor family member; its expression is rapidly induced in cells treated with TGF-beta . To ascertain whether TIEG plays a major role in the TGF-beta pathway, human osteosarcoma MG-63 cells were stably transfected either with an expression vector containing a TIEG cDNA or with the vector alone. Clones that contain only the vector express normal levels of TIEG mRNA and protein and display the same patterns of gene expression and levels of cell proliferation as the nontransfected, non-TGF-beta -treated parental cells. However, transfected cells that overexpress TIEG mRNA and protein (TIEG-6 and TIEG-7) display changes that mimic those of MG-63 cells treated with TGF-beta , i.e. increased alkaline phosphatase activity, decreased levels of osteocalcin mRNA and protein, and decreased cell proliferation. The degree of these changes correlated with the level of TIEG expressed in the cell lines. TGF-beta treatment of the overexpressed cells showed no added effects. These findings and other published reports support a primary role of TIEG as a transcription factor in the TGF-beta signaling pathway.

* This work was supported by National Institutes of Health (NIH) Grant AR43627, the Howard Wagner Cancer research Fund, the Mayo Foundation, and NIH Training Grant Awards HD07108 (to G. G. R.) and CA09441 (to K. M. W.).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.

§ These authors contributed equally to this manuscript.

|| To whom correspondence should be addressed: Dept. of Biochemistry and Molecular Biology, 1601A Guggenheim Bldg., Mayo Clinic, 200 First Street S.W., Rochester, MN 55905. Tel.: 507-284-2480; Fax: 507-284-2053; E-mail: spelsberg.thomas@mayo.edu.

Copyright © 2000 by The American Society for Biochemistry and Molecular Biology, Inc.
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