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Published on: May 27, 2016
Cloning and characterization of the BRD7 gene promoter
Huaying Liu1, Cong Peng, Ming Zhou
1Cancer Research Institute, Xiang-Ya School of Medicine, Central South University, Hunan, People's Republic of China.
Abstract:
BRD7, a novel bromodomain gene, encodes a protein that inhibits cell growth and cell cycle progression by transcriptional regulation of some cell cycle-related genes. Its transcriptional down-expression has been shown to be critical to the pathogenesis of Nasopharyngeal carcinoma (NPC). Little is known about the transcriptional mechanisms controlling BRD7 gene expression. In this paper, we have characterized the 5' regulatory region of the BRD7 gene in order to understand the molecular mechanisms regulating its expression. Transient transfection results suggested that the analyzed upstream sequences of the BRD7 gene might contain some important but not sufficient sequence information to confer the cell-type specificity of BRD7 gene expression. Further analysis with a series of deletions demonstrated that a 125-bp region was required for the basal promoter activity of the BRD7 gene. Results from ChIP and EMSA indicated that the promoter was responsive to Sp1, E2F, and E2F6. All of these suggest a possible mechanism that transcriptional factor Sp1, E2F, and E2F-6 are associated in the BRD7 promoter region and regulate BRD7 promoter activity. Taken together, these results will help to better understand the role of the BRD7 gene in signal-dependent transcriptional regulation, and to develop new reagents for therapeutic upregulation of the BRD7 gene in NPC.
Insights
Bromodomain gene 7 (BRD7) inhibits cell growth and its reduced expression is critical in Nasopharyngeal carcinoma (NPC). This study identified a 125-bp region in the BRD7 promoter regulated by Sp1, E2F, and E2F6 transcription factors.
Area of Science:
- Molecular Biology
- Cancer Genetics
Background:
- Bromodomain gene 7 (BRD7) plays a role in inhibiting cell growth and cell cycle progression.
- Down-expression of BRD7 is implicated in the pathogenesis of Nasopharyngeal carcinoma (NPC).
- The transcriptional mechanisms regulating BRD7 gene expression remain largely unknown.
Purpose of the Study:
- To characterize the 5' regulatory region of the BRD7 gene.
- To elucidate the molecular mechanisms governing BRD7 gene expression.
- To identify key regulatory elements and transcription factors involved in BRD7 promoter activity.
Main Methods:
- Transient transfection assays to assess promoter activity.
- Deletion analysis to identify essential promoter regions.
- Chromatin immunoprecipitation (ChIP) and Electrophoretic Mobility Shift Assay (EMSA) to investigate transcription factor binding.
Main Results:
- Upstream sequences of BRD7 contain elements for cell-type specificity, but are not sufficient alone.
- A 125-bp region is essential for basal promoter activity of the BRD7 gene.
- The BRD7 promoter is responsive to transcription factors Sp1, E2F, and E2F6, suggesting their cooperative role.
Conclusions:
- Sp1, E2F, and E2F6 are likely associated with the BRD7 promoter region, regulating its activity.
- Understanding these regulatory mechanisms is crucial for comprehending BRD7's role in signal-dependent transcription.
- These findings may facilitate the development of therapeutic strategies for NPC by targeting BRD7 upregulation.
