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A Transgenic Analysis of Mouse Lactate Dehydrogenase c Promoter Activity in the Testis

TIM L. KROFT^*,

SIMING LI^*,

LYNN DOGLIO

From the ^* Department of Biochemistry, Molecular Biology, and Cell Biology, Northwestern University, Evanston, and Northwestern University Medical School and the Children's Memorial Institute for Education and Research, Chicago, Illinois. Present address: Department of Biology, Emory University, 1510 Clifton Rd, Atlanta, GA 30322. Present address: Dana-Farber Cancer Institute and Department of Genetics, Harvard Medical School, Boston, MA 02115.

Correspondence to: Erwin Goldberg, Department of Biochemistry, Molecular Biology, and Cell Biology, Northwestern University, 2153 Sheridan Road, Evanston, IL 60208 (fax: 847-467-1380; e-mail: erv{at}northwestern.edu).

Transcription of the mouse testis-specific lactate dehydrogenase c (mldhc) gene is limited to cells of the germinal epithelium. Cloning and analysis of the mldhc promoter revealed that a 100-bp core promoter was able to regulate testis-specific transcription in vitro and in transgenic mice. Surprisingly, expression of the reporter in transgenic testes was limited to pachytene spermatocytes, whereas native LDH-C₄ was detected in pachytene and all later germ cells. To locate additional regulatory sequence that could recapitulate the native LDH-C₄ distribution pattern, we investigated the contribution of 5' and 3' flanking sequences to the regulation of LDH-C₄ expression. We found that transcription factor YY1 binds to the mldhc promoter, that the mldhc 3' untranslated sequence does not permit a postmeiotic expression of a ß-galactosidase reporter in transgenic mice, and that native mldhc mRNA is predominately meiotic, with only a low level of postmeiotic distribution. Our results suggest that the high level of LDH-C₄ in postmeiotic cells results from mRNA and protein stability.

     Key words: Transcription, mRNA stability, protein stability

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