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Volume 271, Number 50, Issue of December 13, 1996 pp. 32375-32380
©1996 by The American Society for Biochemistry and Molecular Biology, Inc.

Transcription of the Transforming Growth Factor-beta 2 Gene Is Dependent on an E-box Located between an Essential cAMP Response Element/Activating Transcription Factor Motif and the TATA Box of the Gene

(Received for publication, July 12, 1996, and in revised form, September 12, 1996)

Beáta Scholtz par , Michelle Kingsley-Kallesen par and Angie Rizzino par

From the  Eppley Institute for Research in Cancer and Allied Diseases, par  Department of Pathology and Microbiology, University of Nebraska Medical Center, Omaha, Nebraska 68198-6805

Transforming growth factor-beta 2 (TGF-beta 2) is an important regulator of cell proliferation and differentiation; however, its transcriptional regulation is not well understood. Here we report characterization of an essential E-box motif, positioned at -50/-45 between a previously described functional cAMP response element/activating transcription factor site and the TATA box of the human TGF-beta 2 promoter. By site-directed mutagenesis, we demonstrate that this E-box motif is necessary for the promoter activity, not only in differentiated cells derived from embryonal carcinoma cells, but also in choriocarcinoma cells and in MCF-7 breast carcinoma cells. We also demonstrate that the transcription factors USF1 and USF2 bind to this E-box motif in vitro when nuclear extracts from each of these cell lines are examined by gel retardation assays. Moreover, using a dominant-negative USF2 protein, we show that USF proteins are critical for TGF-beta 2 promoter activity in vivo. The importance of the E-box motif described in this study is supported by the presence of an E-box motif in the same position in the chicken TGF-beta 2 gene promoter.


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