Okawa Y, Adachi A, Ino M, Nagumo S, Matsui H
Department of Neuropsychiatry, St. Marianna University School of Medicine, Kawasaki, Kanagawa, Japan.
Psychiatry Clin Neurosci. 1998 Aug;52(4):453-8. doi: 10.1046/j.1440-1819.1998.00404.x.
Further characterization of the 5'-flanking promoter region of the human beta1-adrenergic receptor (AR) gene was attempted. The transcription initiation sites, determined by the primer extension and the rapid amplification of the 5'-cDNA end, are multiple in a spanning about 30 nucleotides (-289 to -261 relative to the translation start site). There exist inverted CCAAT boxes, multiple binding sites for transcription factor Sp1 and AP-2 nearby transcription initiation sites, however, this region lacks a typical TATA box. In order to localize the regulatory region for the basal transcription of the human beta1-AR gene, a variety of 5'-flanking sequence/chloramphenicol acetyltransferase reporter gene fusion constructs was prepared and transiently expressed in HeLa cells. Functional analyses reveal negatively (-3813 to -2925 and -1772 to -796) as well as positively (-2925 to -1772) regulatory regions, in addition to the region (-796 to -87) being necessary for the basic expression of the human beta1-AR gene.
我们尝试对人β1 - 肾上腺素能受体（AR）基因的5' - 侧翼启动子区域进行进一步表征。通过引物延伸和5' - cDNA末端快速扩增确定的转录起始位点在跨越约30个核苷酸（相对于翻译起始位点为 - 289至 - 261）的范围内有多个。在转录起始位点附近存在反向CCAAT框、转录因子Sp1和AP - 2的多个结合位点，然而，该区域缺乏典型的TATA框。为了定位人β1 - AR基因基础转录的调控区域，制备了多种5' - 侧翼序列/氯霉素乙酰转移酶报告基因融合构建体，并在HeLa细胞中瞬时表达。功能分析揭示了负向（ - 3813至 - 2925和 - 1772至 - 796）以及正向（ - 2925至 - 1772）调控区域，此外，人β1 - AR基因基础表达所需的区域为（ - 796至 - 87）。
