Demonstration of a testis-specific trans-acting factor Tet-1 in vitro that binds to the promoter of the mouse protamine 1 gene.
We have established testis-specific in vitro transcription of the mouse protamine 1 (MP1) gene using rat testis nuclear extracts. Addition of testis nuclear extracts to brain extracts enhanced transcription from the MP1 upstream sequence-carrying adenovirus major late promoter. Moreover, the MP1 upstream region from positions -92 to -41 alone exhibited transcriptional activation in a tissue-specific manner. DNase I footprinting demonstrated the presence of a DNA-binding factor around position -60 (Tet-1) in testis nuclear extracts, but not in other tissues. Gel shift analysis also revealed the presence of testis-specific Tet-1. Since mutational analysis in transcriptional and binding assays demonstrates that the Tet-1 site is responsible for transcriptional activation, we suggest that Tet-1 is a novel tissue-specific trans-acting factor. The Tet-1-recognizing sequence was delineated to the 11-mer TGACTTCATAA at position -64. Although the first 8-mer in the Tet-1 11-mer shares homology with the cyclic AMP-responsive element, Tet-1 is demonstrated to be distinct from known cAMP-responsive element-binding factors.
Tamura
T
T
Department of Biochemistry, Saitama Medical School, Japan.
Makino
Y
Y
Mikoshiba
K
K
Muramatsu
M
M
D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D001483:Base Sequence; D004247:DNA / Q000235:genetics; D016172:DNA Fingerprinting; D004268:DNA-Binding Proteins / Q000378:metabolism; D005260:Female; D005786:Gene Expression Regulation; D005814:Genes, Viral; D008297:Male; D051379:Mice; D008969:Molecular Sequence Data; D009154:Mutation; D010957:Plasmids; D011401:Promoter Regions, Genetic; D011479:Protamines / Q000235:genetics; D013379:Substrate Specificity; D013737:Testis / Q000378:metabolism*; D015534:Trans-Activators / Q000378:metabolism*; D014157:Transcription Factors / Q000235:genetics; D014158:Transcription, Genetic
D004268:DNA-Binding Proteins; D011479:Protamines; D015534:Trans-Activators; D014157:Transcription Factors; D004247:DNA