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Transcriptional regulation of the hu...
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Zhao, Xin.
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Transcriptional regulation of the human angiotensin II type 1 receptor gene.
紀錄類型:
書目-語言資料,印刷品 : Monograph/item
正題名/作者:
Transcriptional regulation of the human angiotensin II type 1 receptor gene./
作者:
Zhao, Xin.
面頁冊數:
129 p.
附註:
Adviser: Terry S. Elton.
Contained By:
Dissertation Abstracts International62-06B.
標題:
Chemistry, Biochemistry. -
電子資源:
http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=3017336
ISBN:
049327782X
Transcriptional regulation of the human angiotensin II type 1 receptor gene.
Zhao, Xin.
Transcriptional regulation of the human angiotensin II type 1 receptor gene.
- 129 p.
Adviser: Terry S. Elton.
Thesis (Ph.D.)--Brigham Young University, 2001.
The peptide hormone, angiotensin II, regulates a variety of physiological responses which are mediated by its interaction with high affinity G protein-coupled receptors localized on the surface of target cells. This study was initiated to better define the regulation of the human angiotensin II type 1 receptor (hAT1R) gene at the transcriptional level. In this study, the functional promoter of the hAT1R gene was identified and the transcription initiation site was located to an “A” residue 24 by downstream from the putative TATA box. The TATA box has influence on promoter activity and the regulatory mechanism was cell specific. The promoter activity experiment demonstrated that a 145 base pair (bp) sequence within the promoter region was required for basal level expression of the hAT1R gene in various cell lines. Deletion analysis of the hAT1R promoter localized the major regulatory region to the GC-box at −105 to −85 by relative to the transcription start site in H295-R cells. Electrophoretic mobility shift assays (EMSAs) using double-stranded (ds-) oligonucleotide corresponding to this region and H295-R cell nuclear extract resulted in five DNA-protein complexes. EMSAs performed with competitive ds-oligonucleotides which harbored the consensus binding site for Sp1 prevented the formation of the DNA-protein complexes. Super-shift EMSAs also demonstrated that Sp1 and Sp3 could bind to the GC-box present within the −105 to −85 by region of the hAT<sub>1</sub>R promoter. Transactivation experiments utilizing Drosophila SL2 cells, which lack endogenous Sp family transcription factors, demonstrated that Sp1 and Sp3 activated the hAT<sub>1</sub>R promoter. Taken together, these findings suggest that the −105 to −85 by region of the hAT<sub>1</sub>R promoter is important for binding of transcription factors Sp1 and Sp3 and are necessary for the expression of the hAT<sub>1</sub>R gene in H295R cells.
ISBN: 049327782XSubjects--Topical Terms:
1017722
Chemistry, Biochemistry.
Transcriptional regulation of the human angiotensin II type 1 receptor gene.
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The peptide hormone, angiotensin II, regulates a variety of physiological responses which are mediated by its interaction with high affinity G protein-coupled receptors localized on the surface of target cells. This study was initiated to better define the regulation of the human angiotensin II type 1 receptor (hAT1R) gene at the transcriptional level. In this study, the functional promoter of the hAT1R gene was identified and the transcription initiation site was located to an “A” residue 24 by downstream from the putative TATA box. The TATA box has influence on promoter activity and the regulatory mechanism was cell specific. The promoter activity experiment demonstrated that a 145 base pair (bp) sequence within the promoter region was required for basal level expression of the hAT1R gene in various cell lines. Deletion analysis of the hAT1R promoter localized the major regulatory region to the GC-box at −105 to −85 by relative to the transcription start site in H295-R cells. Electrophoretic mobility shift assays (EMSAs) using double-stranded (ds-) oligonucleotide corresponding to this region and H295-R cell nuclear extract resulted in five DNA-protein complexes. EMSAs performed with competitive ds-oligonucleotides which harbored the consensus binding site for Sp1 prevented the formation of the DNA-protein complexes. Super-shift EMSAs also demonstrated that Sp1 and Sp3 could bind to the GC-box present within the −105 to −85 by region of the hAT<sub>1</sub>R promoter. Transactivation experiments utilizing Drosophila SL2 cells, which lack endogenous Sp family transcription factors, demonstrated that Sp1 and Sp3 activated the hAT<sub>1</sub>R promoter. Taken together, these findings suggest that the −105 to −85 by region of the hAT<sub>1</sub>R promoter is important for binding of transcription factors Sp1 and Sp3 and are necessary for the expression of the hAT<sub>1</sub>R gene in H295R cells.
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