Nuclear factor-Y (NF-Y) regulates transcription of mouse Dmrt7 gene by binding to tandem CCAAT boxes in its proximal promoter
<p><i>Dmrt7</i>, a member of the <i>Dmrt</i> family of genes, is required for spermatogenesis. However, promoter functions of the gene<i> Dmrt7 </i>remain unknown. We have cloned and characterized the proximal promoter region of the mouse <i>Dmrt7</...
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| Format: | Article |
| Language: | English |
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Ivyspring International Publisher
2010-01-01
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| Series: | International Journal of Biological Sciences |
| Online Access: | http://www.biolsci.org/v06p0655.htm |
| Summary: | <p><i>Dmrt7</i>, a member of the <i>Dmrt</i> family of genes, is required for spermatogenesis. However, promoter functions of the gene<i> Dmrt7 </i>remain unknown. We have cloned and characterized the proximal promoter region of the mouse <i>Dmrt7</i> gene. Functional analysis of the 5' flanking region by sequential deletion mutations revealed crucial positive elements between -60 and +1, in which two highly conserved and tandem CCAAT boxes: the CCAAT box1 (-48/-44) and the CCAAT box2 (-7/-3) are located. Site-directed mutagenesis studies demonstrated that both CCAAT boxes are indispensable to the promoter activity. Electrophoretic mobility shift assays (EMSAs) and gel-supershift assays indicated that transcription factor NF-Y binds to the promoter. Chromatin immunoprecipitation (ChIP) analysis demonstrated that NF-Y interacts <i>in vivo</i> with the promoter of the <i>Dmrt7</i> gene in testis. Co-transfection and reporter analysis showed that over-expression of NF-Ys increased transcription of the <i>Dmrt7-luc </i>gene whereas expression of a dominant-negative NF-Ya decreased the transcription. This suggests that NF-Y can activate the <i>Dmrt7</i> promoter. These results provide evidence of a transcription regulatory mechanism that controls <i>Dmrt7</i> gene expression in mouse testis.</p> |
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| ISSN: | 1449-2288 |