Deletional analysis of the human renin promoter: transcriptional activation by the SV40 enhancer enables identification of promoter regulatory elements in non-renin-expressing cells.
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.
Kasahara
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Department of Endocrinology, Tokyo Medical and Dental University.
D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics; D004274:DNA, Recombinant; D004742:Enhancer Elements, Genetic; D015971:Gene Expression Regulation, Enzymologic; D005814:Genes, Viral; D005819:Genetic Markers; D006367:HeLa Cells; D006801:Humans; D009928:Organ Specificity; D011401:Promoter Regions, Genetic; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics; D012083:Renin / Q000096:biosynthesis / Q000235:genetics*; D012260:Ribonucleases / Q000032:analysis; D017384:Sequence Deletion; D013539:Simian virus 40 / Q000235:genetics; D014158:Transcription, Genetic; D014162:Transfection
D004274:DNA, Recombinant; D005819:Genetic Markers; D011993:Recombinant Fusion Proteins; D015500:Chloramphenicol O-Acetyltransferase; D012260:Ribonucleases; D012083:Renin