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A Short Core Promoter Drives Expression of the ALF Transcription Factor in Reproductive Tissues of Male and Female Mice¹

Department of Molecular and Cell Biology,³ The University of Texas at Dallas, Richardson, Texas 75080 Department of Anatomy and Cell Biology,⁴ McGill University School of Medicine, Strathcona Building, Montreal, Quebec H3A 2B2, Canada Department of Physiology,⁵ West China Center of Medical Sciences, Sichuan University, Chengdu 610041, Peoples Republic of China

The control of gene expression in reproductive tissues involves a number of unique germ cell-specific transcription factors. One such factor, ALF (TFIIA), encodes a protein similar to the large subunit of general transcription factor TFIIA. To understand how this factor is regulated, we characterized transgenic mice that contain the ALF promoter linked to either ß-galactosidase or green fluorescent protein (GFP) reporters. The results show that as little as 133 base pairs are sufficient to drive developmentally accurate and cell-specific expression. Transgene DNA was methylated and inactive in liver, but could be reactivated in vivo by system administration of 5-aza, 2'-deoxycytidine. Fluorescence-activated cell sorting allowed the identification of male germ cells that express the GFP transgene and provides a potential method to collect cells that might be under the control of a nonsomatic transcription system. Finally, we found that transcripts from the endogenous ALF gene and derived transgenes can also be detected in whole ovary and in germinal vesicle-stage oocytes of female mice. The ALF sequence falls into a class of germ cell promoters whose features include small size, high GC content, numerous CpG dinucleotides, and an apparent TATA-like element. Overall, the results define a unique core promoter that is active in both male and female reproductive tissues, and suggest mouse ALF may have a regulatory role in male and female gametogenic gene expression programs.

¹ Support for this work was provided by the American Cancer Society, the Robert A. Welch Foundation, and the National Institutes of Health. L.Y. was supported in part by a fellowship from the West China Center of Medical Sciences, Sichuan University. S.Y.H. and W.X. contributed equally to this work.

² Correspondence: Jeff DeJong, The University of Texas at Dallas, Department of Molecular and Cell Biology, 2601 N. Floyd Road, Richardson, TX 75080. FAX: 972 883 2409; dejong{at}utdallas.edu

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