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Volume 272, Number 8, Issue of February 21, 1997 pp. 5056-5062
©1997 by The American Society for Biochemistry and Molecular Biology, Inc.

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Novel Repeat Elements Direct Rat Proenkephalin Transcription during Spermatogenesis

(Received for publication, August 19, 1996, and in revised form, November, 20, 1996)

From the Neurobiology Group, Worcester Foundation for Biomedical Research, Shrewsbury, Massachusetts 01545 and the ^§ Department of Anatomy and Cell Biology, Tufts University School of Veterinary Medicine, North Grafton, Massachusetts 01536

The developmental program controlling sperm formation occurs in multiple stages that sequentially involve mitosis, meiosis, and spermiogenesis. The transcriptional mechanisms regulating these distinct phases are poorly understood. In particular, while a required role for the germ cell transcription factor cyclic AMP response element modulator- during spermiogenesis has recently been demonstrated, the transcriptional mechanisms leading to early haploid cell formation are unknown. The rat and mouse proenkephalin genes are selectively expressed from an alternate, germ cell-specific promoter in meiotic and early haploid cells. In this study, the minimal rat proenkephalin germ line promoter was localized to a 116-bp region encompassing the transcriptional start site region. Further, a proximal 51-bp sequence located in the 5-flanking region is absolutely required for germ line promoter activity. This 51 bp sequence corresponds to a previously characterized binding element (GCP1) that forms cell-specific complexes with rat spermatogenic cell nuclear factors distinct from cyclic AMP response element binding proteins. Further, GCP1 contains novel direct repeat sequences required for factor binding and transgene expression in spermatogenic cells. These repeat elements are highly similar to sequences within the active regions of other male germ line promoters expressed during meiosis. GCP1 may therefore contain transcriptional elements that participate more generally during meiosis in the differentiation of spermatocytes and early haploid spermatids.

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