JBC Transcription and Nuclear Factor Monoclonals

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J Biol Chem, Vol. 275, Issue 11, 8114-8120, March 17, 2000

Yin-yang 1 and Glucocorticoid Receptor Participate in the Stat5-mediated Growth Hormone Response of the Serine Protease Inhibitor 2.1 Gene*

Pearl L. BergadDagger , Howard C. Towle§, and Susan A. BerryDagger ||

From the Departments of Dagger  Pediatrics and § Biochemistry, Molecular Biology, and Biophysics and the  Institute of Human Genetics, University of Minnesota, Minneapolis, Minnesota 55455

A growth hormone-inducible nuclear factor complex (GHINF), affinity-purified using the growth hormone response element (GHRE) from the promoter of rat serine protease inhibitor 2.1, was found to contain Stat5a and -5b, as well as additional components. The ubiquitous transcription factor yin-yang 1 (YY1) is present in GHINF. An antibody to YY1 inhibited the formation of the GHINF·GHRE complex in an electrophoretic mobility shift assay. Furthermore, Stat5 was co-immunoprecipitated from rat hepatic nuclear extracts with antibodies to YY1. An examination of the GHRE shows that, in addition to two gamma -activated sites, it contains a putative YY1 binding site between the two gamma -activated sites, overlapping them both. Mutation of this putative YY1 site results in a decrease of GHINF·GHRE complex formation in an electrophoretic mobility shift assay and a corresponding decrease in growth hormone (GH) response in functional assays. The glucocorticoid receptor was also present in GHINF, and Stat5 co-immunoprecipitates with glucocorticoid receptor in hepatic nuclear extracts from rats treated with GH. GH activation of serine protease inhibitor 2.1 requires the unique sequence of the GHRE encompassing the recognition sites of several transcription factors, and the interaction of these factors enhances the assembly of the transcription complex.

* This work was supported by National Institutes of Health Grant DK32817, the Genentech Foundation for Growth and Development, and the Vikings Children's Fund.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.

|| To whom correspondence should be addressed: Dept. of Pediatrics, University of Minnesota, 420 Delaware St. S.E., Box 75, Minneapolis, MN 55455. Tel.: 612-624-7144; Fax: 612-624-2682; E-mail: berry002@tc.umn.edu.

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