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BOR - Papers in Press, published online ahead of print May 12, 2004.
Biol Reprod 2004, 10.1095/biolreprod.104.028969
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BIOLOGY OF REPRODUCTION 71, 837–844 (2004)
DOI: 10.1095/biolreprod.104.028969
© 2004 by the Society for the Study of Reproduction, Inc.


Testis

Ontogeny of a Demethylation Domain and Its Relationship to Activation of Tissue-Specific Transcription1

Christopher B. Geyer3, Christine Mione Kiefer4, Thomas P. Yang4,6, and John R. McCarrey2,3,7

Department of Cellular and Structural Biology,3 University of Texas Health Science Center at San Antonio, San Antonio, Texas 78229 Department of Biochemistry and Molecular Biology,4 Center for Mammalian Genetics,5 Department of Pediatrics,6 University of Florida School of Medicine, Gainesville, Florida 32611 Department of Biology,7 University of Texas at San Antonio, San Antonio, Texas 78249

We examined DNA methylation throughout the endogenous murine testis-specific phosphoglycerate kinase (Pgk2) gene and in human PGK2 promoter/CAT reporter transgenes in mouse spermatogenic cells before, during, and following the period of active transcription of this gene. We observed the gradual development of a domain of demethylation beginning over the promoter and then expanding approximately 1 kilobase in each direction within the endogenous Pgk2 gene. This demethylation domain develops in the absence of DNA replication and precedes other molecular changes that potentiate tissue-specific activation of this gene. Studies with transgenes show that a signal residing in the Pgk2 core promoter directs this gene-, cell type-, and stage-specific demethylation process. These results are consistent with a model in which regulated, tissue- and gene-specific demethylation initiates a cascade of subsequent molecular events required for tissue-specific activation of transcription during spermatogenesis in vivo.

1 Supported by a grant from the NIH to J.R.M. (HD 40891).

2 Correspondence. FAX: 210.458.5658; jmccarrey{at}utsa.edu




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