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Molecular and Cellular Biology, January 2004, p. 377-388, Vol. 24, No. 1
0270-7306/04/$08.00+0     DOI: 10.1128/MCB.24.1.377-388.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Gata4 Regulates Testis Expression of Dmrt1

Ning Lei and Leslie L. Heckert*

Department of Molecular and Integrative Physiology, The University of Kansas Medical Center, Kansas City, Kansas 66160

Received 27 December 2002/ Returned for modification 18 February 2003/ Accepted 30 September 2003

The doublesex and mab-3 related transcription factor 1 (Dmrt1) is a putative transcriptional regulator that is expressed exclusively in the gonads and is required for postnatal testis differentiation. Here we describe the transcriptional mechanisms regulating testis-specific expression of the Dmrt1 gene. Transient-transfection analysis identified a region of the promoter between kb -3.2 and -2.8 that is important for Sertoli cell-specific expression. DNase I footprinting revealed four sites of DNA-protein interaction within this region, three of which were prominent in primary Sertoli cells. Analysis of these sites, using electrophoretic mobility shift assays, revealed that Gata4 and another unknown factor bound within these regions. Further transient-transfection assays of various mutant promoters established the functional relevance of the Gata4-response and unknown factor-response elements, while studies of Dmrt1 expression in 13.5 days postcoitum Fog2 null gonads supported the in vivo importance of Gata4's regulation. As a whole, these studies identify Gata4 as an important regulator in the Dmrt1 transcriptional machinery that is responsible for robust expression of Dmrt1 in the testis.

* Corresponding author. Mailing address: Department of Molecular and Integrative Physiology, The University of Kansas Medical Center, 3901 Rainbow Blvd., Kansas City, KS 66160. Phone: (913) 588-7588. Fax: (913) 588-7530. E-mail: lheckert{at}kumc.edu.

Molecular and Cellular Biology, January 2004, p. 377-388, Vol. 24, No. 1
0022-538X/04/$08.00+0     DOI: 10.1128/MCB.24.1.377-388.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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