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Originally published In Press as doi:10.1074/jbc.M003345200 on June 26, 2000

J. Biol. Chem., Vol. 275, Issue 36, 27989-27999, September 8, 2000
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Heterodimeric Pbx-Prep1 Homeodomain Protein Binding to the Glucagon Gene Restricting Transcription in a Cell Type-dependent Manner*

Stephan HerzigDagger , Laszlo Füzesi§, and Willhart KnepelDagger

From the Dagger  Department of Molecular Pharmacology and § Department of Gastroenteropathology, University of Göttingen, 37075 Göttingen, Germany

Homeodomain proteins specify developmental pathways and cell-specific gene transcription whereby proteins of the PBC subclass can direct target gene specificity of Hox proteins. Proteins encoded by nonclustered homeobox genes have been shown to be essential for cell lineage differentiation and gene expression in pancreatic islets. Using specific antiserum in an electrophoretic mobility shift assay and in vitro transcribed/translated proteins, the nuclear proteins binding domain B of the G3 enhancer-like element of the glucagon gene were identified in the present study as heterodimers consisting of the ubiquitously expressed homeodomain protein Prep1 and the also widely expressed PBC homeoprotein Pbx (isoform 1a, 1b, or 2). These heterodimeric complexes were found to bind also to the glucagon cAMP response element and to a newly identified element termed G5 (from -169 to -140). Whereas the expression of Prep1 or Pbx forms alone had no effect, coexpression of Pbx1a/1b-Prep1 inhibited the glucagon promoter when activated by cotransfected Pax6 or another transcription factor in non-glucagon-producing cells. In contrast, in glucagon-producing pancreatic islet cells, Pbx-Prep1 had no effect on GAL4-Pax6-induced mutant glucagon promoter activity or on Pax6-dependent wild-type glucagon promoter activity. Furthermore, 5'-deletion of G5 enhanced glucagon promoter activity in a non-glucagon-producing cell line but not in glucagon-producing islet cells. This study thus identifies a novel target and Hox-independent function of Pbx-Prep1 heterodimers that, through repression of glucagon gene transcription in non-glucagon-producing cells, may help to establish islet cell-specific expression of the glucagon gene.

* This work was supported by Deutsche Forschungsgemeinschaft Grant SFB 402/A3.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 Molecular Pharmacology, University of Göttingen, Robert-Koch-Str. 40, Postfach 3742, D-37070 Göttingen, Germany. Tel.: 49-551-395787; Fax: 49-551-399652; E-mail: wknepel@med.uni-goettingen.de.

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