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

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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