title: Transient expression analyses of DNA extending 2.4 kb upstream of the human renin gene.
abstract: The influence on homologous and heterologous promoter activity of DNA extending 2.4 kb upstream of the human renin gene (REN) was examined by transient expression assay in JEG-3 cells, using the gene for chloramphenicol acetyl transferase (CAT) as reporter, and cotransfection with pCH110 to control for transfection efficiency. Analyses of constituent subfragments of the region 5' of residue -144, using the herpes simplex virus thymidine kinase (tk) promoter to drive transcription, provided no evidence for negative regulatory influences within the -2400 to -144 DNA. That distal 5'-flanking DNA may have little influence on promoter activity is further supported by a sharp decline in nucleotide homology between human, rat and mouse renin genes further upstream than human residue -604. Constructs containing renin DNA to residue +13, i.e., which retained the REN promoter, all displayed very low CAT activities, consistent with negative cis-acting control within the -149 to +13 region. This finding contrasts with results of similar studies for mouse, in which renin gene control was suggested to be mediated primarily via cell-specific trans-acting activator(s) acting on yet-to-be identified enhancer(s). Mouse renin genes have, however, a common DNA insertion that could have disrupted the negative element in this region, and which might contain enhancer target(s) for trans-acting factor(s). In conclusion, the present study involving JEG-3 cells has demonstrated that distal human renin 5'-flanking DNA has little cis-acting influence on promoter activity, whereas DNA located within 100 base pairs of the renin promoter may have a negative regulatory effect.(ABSTRACT TRUNCATED AT 250 WORDS)
authors.affiliation: Department of Physiology, University of Sydney, NSW, Australia.
authors.forename: D L
authors.initials: DL
authors.lastname: Smith
authors.forename: B J
authors.initials: BJ
authors.lastname: Morris
mesh_terms: D003001:Cloning, Molecular; D015870:Gene Expression; D006801:Humans; D011401:Promoter Regions, Genetic; D012083:Renin / Q000235:genetics*; D015183:Restriction Mapping; D012689:Sequence Homology, Nucleic Acid; D014407:Tumor Cells, Cultured
chemical_list: D012083:Renin