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人生长激素促分泌素受体基因5'侧翼区的克隆与特性分析

Cloning and characterization of the 5'-flanking region of the human growth hormone secretagogue receptor gene.

作者信息

Kaji H, Tai S, Okimura Y, Iguchi G, Takahashi Y, Abe H, Chihara K

机构信息

Third Division, Department of Medicine, Kobe University School of Medicine, Kobe 650-0017, Japan.

出版信息

J Biol Chem. 1998 Dec 18;273(51):33885-8. doi: 10.1074/jbc.273.51.33885.

Abstract

Recently, the growth hormone secretagogue receptor (GHS-R) cDNA has been isolated from the pituitary and hypothalamus. To evaluate the regulation of human (h) GHS-R gene expression, we cloned the hGHS-R gene containing the 5'-flanking region of 0.6-2.9 kilobase pairs. Analysis of the hGHS-R transcripts with 5'-rapid amplification of cDNA ends suggested that the putative transcription initiation site was approximately -453 base pairs upstream of the translation initiation site (+1). There is no typical TATA, CAAT, or GC box but an initiator-like sequence and putative binding sites for several transcription factors around the putative transcription start site. The 5'-flanking region inserted into a luciferase reporter vector had promoter activity in GH3 cells but had activity indistinguishable from background in HeLa or EP1 cells. The hGHS-R promoter activity in GH3 cells increased by deletion of nucleotides from -1224 to -734, whereas it was decreased by further deletion from -734 to -608. Knowledge of the promoter region of the hGHS-R gene will facilitate elucidation of its transcriptional control.

摘要

最近,已从垂体和下丘脑分离出了生长激素促分泌素受体(GHS-R)cDNA。为了评估人类(h)GHS-R基因表达的调控,我们克隆了包含0.6 - 2.9千碱基对5'侧翼区域的hGHS-R基因。用5' cDNA末端快速扩增法分析hGHS-R转录本表明,推定的转录起始位点在翻译起始位点(+1)上游约453个碱基对处。在推定的转录起始位点周围没有典型的TATA、CAAT或GC盒,但有一个类似起始子的序列以及几个转录因子的推定结合位点。插入荧光素酶报告载体的5'侧翼区域在GH3细胞中具有启动子活性,但在HeLa或EP1细胞中的活性与背景无差异。通过从 - 1224至 - 734缺失核苷酸,GH3细胞中的hGHS-R启动子活性增加,而从 - 734至 - 608进一步缺失则使其活性降低。了解hGHS-R基因的启动子区域将有助于阐明其转录调控机制。

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