Castrén M, Damm K
Max-Planck-Institute of Psychiatry, Department of Neuroendocrinology, München, Germany.
J Neuroendocrinol. 1993 Aug;5(4):461-6. doi: 10.1111/j.1365-2826.1993.tb00508.x.
The authors have identified two types of hippocampal cDNAs for the rat mineralocorticoid receptor (rMR) which are identical in the protein coding domain but differ in their 5'-untranslated sequences. One of these clones encodes a novel type of rMR cDNA with a high homology to a previously described human MR cDNA isolated from the kidney. A genomic clone containing the 5'-end of the rat MR gene was isolated. The 12.7 kb genomic region contains the 5'-coding exon with the translational start site and contiguous DNA sequences encoding the N-terminal domain of the rMR. A 240 bp region homologous to the 5'-untranslated sequences of the novel rMR cDNA was located 5.2 kb upstream the protein coding region. Characterization of the nucleotide sequence preceding this exon revealed several features characteristic for promoters of so-called 'housekeeping genes'. The sequence analyzed is 635 bp in length, is rich in G+C nucleotides (63%) and lacks TATA or CAAT regulatory elements. It contains three putative binding sites for transcription factor Sp1 as well as several short sequences that are similar to known cis-acting enhancers or binding sites for transcription factors. In transient transfection experiments employing the luciferase reporter gene and the CV1 cell line this region exhibits substantial promoter activity. These experiments demonstrate that expression of the rat MR gene in the hippocampus results in at least two transcripts with different 5'-untranslated exons.(ABSTRACT TRUNCATED AT 250 WORDS)
作者们鉴定出了大鼠盐皮质激素受体(rMR)的两种海马cDNA类型,它们在蛋白质编码结构域相同,但5'-非翻译序列不同。其中一个克隆编码一种新型的rMR cDNA,与先前从肾脏分离出的人类MR cDNA具有高度同源性。分离出了一个包含大鼠MR基因5'-末端的基因组克隆。这个12.7 kb的基因组区域包含带有翻译起始位点的5'-编码外显子以及编码rMR N-末端结构域的相邻DNA序列。在蛋白质编码区域上游5.2 kb处定位到了一个与新型rMR cDNA的5'-非翻译序列同源的240 bp区域。对该外显子之前的核苷酸序列进行特征分析,发现了一些所谓“管家基因”启动子的特征。所分析的序列长度为635 bp,富含G+C核苷酸(63%),并且缺乏TATA或CAAT调控元件。它包含三个转录因子Sp1的假定结合位点以及几个与已知顺式作用增强子或转录因子结合位点相似的短序列。在使用荧光素酶报告基因和CV1细胞系的瞬时转染实验中,该区域表现出显著的启动子活性。这些实验表明,大鼠MR基因在海马中的表达产生了至少两种具有不同5'-非翻译外显子的转录本。(摘要截短于250字)