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BOR - Papers in Press, published online ahead of print January 22, 2003.
Biol Reprod 2003, 10.1095/biolreprod.102.014084
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BIOLOGY OF REPRODUCTION 68, 2267–2273 (2003)
DOI: 10.1095/biolreprod.102.014084
© 2003 by the Society for the Study of Reproduction, Inc.

Testis

Specific Binding of Nuclear Proteins to a Bifunctional Promoter Element Upstream of the H1/AC Box of the Testis-Specific Histone H1t Gene1

Steven A. Wolfe, and Sidney R. Grimes2

Research Service (151), Overton Brooks Veterans Administration Medical Center, Shreveport, Louisiana 71101-4295 Department of Biochemistry and Molecular Biology, Louisiana State University Health Science Center, Shreveport, Louisiana 71130-3932

The testis-specific histone H1t gene is transcribed exclusively in primary spermatocytes during spermatogenesis. Studies with transgenic mice show that 141 base pairs (bp) of the H1t proximal promoter accompanied with 800 bp of downstream sequence are sufficient for tissue-specific transcription. Nuclear proteins from testis and pachytene spermatocytes produce footprints spanning the region covering the repressor element (RE) from 100 to 125 nucleotides upstream of the H1t transcriptional initiation site. Only testis nuclear proteins bind to the 5'-end of the element and produce a unique, low-mobility complex in electrophoretic mobility shift assays. This testis complex is distinct from the complex formed by a repressor protein derived from several cell lines that binds to the 3'-end of the element. The testis complex band is formed when using nuclear proteins from primary spermatocytes, where the H1t gene is transcribed, and band intensity drops 70%–80% when using nuclear proteins from early spermatids, where H1t gene transcription ceases. Protein-DNA cross-linking experiments using testis nuclear proteins produce electrophoretic bands of 59, 52, and 50 kDa on SDS/PAGE gels.

1 Supported by a merit review grant from the Department of Veterans Affairs and by NIH Grant HD29381 (S.R.G.).

2 Correspondence: Sidney R. Grimes, Medical Research Service (151), Overton Brooks Veterans Administration Medical Center, 510 E. Stoner Ave., Shreveport, Louisiana 71101-4295. FAX: 318 429 5733; srgrimes{at}prysm.net




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R. Chowdhary, R. A. Ali, W. Albig, D. Doenecke, and V. B Bajic
Promoter modeling: the case study of mammalian histone promoters
Bioinformatics, June 1, 2005; 21(11): 2623 - 2628.
[Abstract] [Full Text] [PDF]


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Copyright © 2003 by the Society for the Study of Reproduction.