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Distinct Regions of the Mouse Cyclin A1 Gene, Ccna1, Confer Male Germ-Cell Specific Expression and Enhancer Function¹

The Institute of Human Nutrition,³ Departments of Genetics and Development⁴ Obstetrics and Gynecology,⁵ The Center for Reproductive Sciences,⁶ The Herbert Irving Comprehensive Cancer Center,⁷ Columbia University Medical Center, New York, New York 10032

The gene encoding mouse cyclin A1, Ccna1, is expressed at highest levels in late pachytene-diplotene spermatocytes, where it is required for meiotic cell division. To begin to understand the mechanisms responsible for its highly restricted pattern of expression, transgenic mouse lines carrying constructs consisting of the cyclin A1 regulatory region fused with the reporter gene lacZ were generated. Analysis of tissue-specific and testicular cell-type-specific transgene expression indicated that sequences within –1.3 kilobases (kb) of the cyclin A1 putative transcriptional start site were sufficient to direct transgene expression uniquely to late spermatocytes while maintaining repression in other tissues. However, sequences located between –4.8 kb and –1.3 kb of the putative transcriptional start site were apparently required to transcribe the reporter at levels needed for consistent X-gal staining. Comparison of the mouse, rat, and human proximal promoters revealed regions of high sequence conservation and consensus sequences both for known transcription factors, some of which are coexpressed with Ccna1, such as A-myb and Hsf2, and for elements that control expression of genes in somatic cell cycles, such as CDE, CHR, and CCAAT elements. Thus, the promoter region within 1.3 kb upstream of the putative Ccna1 transcriptional start can direct expression of lacZ to spermatocytes, while sequences located between –4.8 kb and –1.3 kb of the putative transcriptional start site may enhance expression of lacZ.

¹ Supported by NIH grants HD34915 (D.J.W) and T32 DK07647 (K.M.L.).

² Correspondence: Debra Wolgemuth, Department of Genetics and Development, Columbia University Medical Center, 630 West 168th Street, Black 1613, New York, NY 10032. FAX: 212 305 6084; djw3{at}columbia.edu

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