Deletional analysis of the human renin promoter: transcriptional activation by the SV40 enhancer enables identification of promoter regulatory elements in non-renin-expressing cells.

N Kasahara
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Abstract

Identification of regulatory elements in the human renin gene promoter has been hindered by the lack of suitable renin-expressing cell lines. In this paper, the SV40 viral enhancer is coupled to a human renin promoter/chloramphenicol acetyltransferase (CAT) reporter gene construct, to determine whether promoter regulatory elements can be identified in the context of enhanced transcription in the normally non-renin-expressing HeLa cell line. The present results indicate that the SV40 enhancer can overcome tissue-specificity of the human renin promoter, and confer correctly initiated transcriptional activity to HeLa cells. Analysis of a series of 5'-end deletions of the human renin promoter linked to the CAT gene and SV40 enhancer identified negative regulatory elements between positions-275 and -225, and between-142 and -102 of the human renin promoter, as well as a positive regulatory element between -225 and -142. Thus, studies of renin promoter regulatory elements need not be limited to renin-expressing cells, but can be performed in non-renin-expressing cells with the addition of an enhancer. This strategy can be generally applicable to the study of tissue-specific gene regulation in cases where there are no specific cell lines available.

人肾素启动子的缺失分析:SV40增强子的转录激活能够识别非肾素表达细胞中的启动子调控元件。
由于缺乏合适的肾素表达细胞系,人类肾素基因启动子调控元件的鉴定一直受到阻碍。本文将SV40病毒增强子与人肾素启动子/氯霉素乙酰转移酶(chloramphenicol acetyltransferase, CAT)报告基因构建体偶联,以确定在正常不表达肾素的HeLa细胞系中是否可以在转录增强的背景下鉴定启动子调控元件。目前的结果表明,SV40增强子可以克服人肾素启动子的组织特异性,并赋予HeLa细胞正确启动的转录活性。对与CAT基因和SV40增强子相关的人肾素启动子的一系列5'端缺失进行分析,发现了人肾素启动子的-275和-225之间、-142和-102之间的负调控元件,以及-225和-142之间的正调控元件。因此,肾素启动子调控元件的研究不必局限于肾素表达细胞,而可以在添加增强子的非肾素表达细胞中进行。在没有特定细胞系的情况下,这种策略可以普遍适用于组织特异性基因调控的研究。
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