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Molecular and Cellular Biology, January 2003, p. 1-13, Vol. 23, No. 1
0270-7306/03/$08.00+0     DOI: 10.1128/MCB.23.1.1-13.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Transcriptional Repression of Peri-Implantation EMX2 Expression in Mammalian Reproduction by HOXA10

Patrick J. Troy, Gaurang S. Daftary, Catherine N. Bagot, and Hugh S. Taylor*

Division of Reproductive Endocrinology, Yale University School of Medicine, New Haven, Connecticut 06520-8063

Received 6 June 2002/ Returned for modification 5 August 2002/ Accepted 4 October 2002

HOXA10 is necessary for mammalian reproduction; however, its transcriptional targets are not completely defined. EMX2, a divergent homeobox gene, is necessary for urogenital tract development. In these studies we identify and characterize the regulation of EMX2 by HOXA10. By using Northern analysis and in situ hybridization, we found that EMX2 is expressed in the adult urogenital tract in an inverse temporal pattern from HOXA10, suggestive of a negative regulatory relationship. Constitutive expression of HOXA10 diminished EMX2 mRNA, whereas blocking HOXA10 through the use of antisense resulted in high EMX2 mRNA expression. Deletional analysis of the EMX2 5' regulatory region revealed that a 150-bp element mediated transcriptional repression when cotransfected with pcDNA3.1/HOXA10 in transient-transfection assays. Binding of HOXA10 protein to this element was demonstrated by electrophoretic mobility shift assay and further localized to a consensus HOXA10 binding site within this element by DNase I footprinting. Site-directed mutagenesis abolished binding, as well as the negative transcriptional regulation. Transcriptional activation of empty spiracles, the Drosophila ortholog of EMX2, by Abdominal-B (HOXA10 ortholog) has been previously demonstrated. These findings demonstrate conservation of the transcription factor-target gene relationship, although the direction of regulation is reversed with possible evolutionary implications.

* Corresponding author. Mailing address: Division of Reproductive Endocrinology, Yale University School of Medicine, 333 Cedar Street, P.O. Box 208063, New Haven, CT 06520-8063. Phone: (203) 785-4005. Fax: (203) 785-7134. E-mail: hugh.taylor{at}yale.edu.

Molecular and Cellular Biology, January 2003, p. 1-13, Vol. 23, No. 1
0022-538X/03/$08.00+0     DOI: 10.1128/MCB.23.1.1-13.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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